Nghiên cứu độc tính và tác dụng của cao lỏng HVT trên hội chứng rối loạn lipid máu tt tiếng anh

1
RATIONALE
Cardiovascular disease has been a major health
concern in the world, with the leading cause of mortality and
disability. According to estimates by the World Health
Organization, there are about 17 million people died from
cardiovascular disease every year, mainly caused by
atherosclerosis.
Dyslipidemia pattern is one of the most important risk
factors but a variable risk factor for the formation and
development of atherosclerosis. Modern medicine has many
drugs used to treat dyslipidemia, but some of them also have
undesirable effects such as muscle pain, muscle cramps,
impaired liver function, etc.
HVT remedy is made up of three commonly used
medicinal herbs: Folium Nelumbinis nuciferae, Cleistocalyx
operculatus Roxb and Pericarpium Citri reticulatae perenne,
are prepared into liquid form which act on improving healthy
qi, resolving phlegm-dampness and fortifying the spleen.
Currently, there are no studies evaluating the effects of
treatment of HVT on dyslipidemia pattern. To demonstrate
the effects and toxicity of HVT, as a scientific basis towards
the ability to widely use this product in clinical practice, we
conduct the research on the topic “A study on the toxicity and
effectiveness of HVT liquid drug on dyslipidemia pattern” with
the following goals:
1. To study the acute toxicity and semi-chronic toxicity of
liquid HVT.
2. To study the effect of dyslipidemia adjustment of liquid
HVT in experimental animals.
3. To evaluate the therapeutic effects and unwanted

effects of liquid HVT in patients with dyslipidemia caused
by internal phlegm-dampness.
PRACTICAL SIGNIFICANCE AND NEW
CONTRIBUTIONS OF THE STUDY
Practical significance
Although the treatment of dyslipidemia with modern
medicine is effective, it also causes some unwanted effects.
Therefore, the search for remedies derived from herbs, with


2
few tastes, effective and safe is always a necessity as well as
a research direction that is of interest to domestic and
international scientists.
Scientific significance
The dissertation has been systematically studied in both
experimental and clinical practice. The drug is produced in
liquid form with ingredients of three traditional medicine
drugs which are originated from Vietnamese traditional
medicinal drugs, easy to find and convenient for use in
treating dyslipidemedia. The study has provided scientific
evidence on the effect of dyslipidemedia adjustment as well
as unwanted effects (if any) on experimental and clinical
issues.
The applied research of a remedy in treatment has
made a contribution to the clarification of the theory of
traditional medicine. At the same time, gradually modernizing
traditional medicine is also a scientific and practical
significance.

ORGANIZATION OF THE STUDY
The dissertation consists of 128 pages, of which: Rationale (2
pages), Literature review (32 pages), Subjects and research
methods (18 pages), Research results (43 pages),
Discussions
(30
pages),
Conclusions
(2
pages),
Recommendations (1 page). There are 145 references used
in the study, of which there are 53 documents in
Vietnamese, 92 documents in English and 2 documents in
Chinese. The dissertation has been presented and illustrated
through 61 tables, 9 charts, 18 pictures, 5 diagrams and
appendices.

Chapter 1. LITERATURE REVIEW
1.1. Dyslipidemia from modern medicine point of view.
1.1.1. Definition of dyslipidemia


3
Dyslipidemia is a condition of the increase of either total
cholesterol (TC) or triglyceride (TG) or both, or the decrease of
high-density lipoprotein cholesterol (HDL-C), or the increase of
low-density lipoprotein cholesterol (LDL-C) which increases the
process of atherosclerosis.
1.1.2. Causes of dyslipidemia
The cause of dyslipidemia can be primary (due to genetic

diseases) or secondary (due to eating habits, lifestyles or some
pathologies). Secondary causes may contribute to
dyslipidemia due to the causes of the primary which become
more severe.
1.1.3. Treatment of dyslipidemia
* Treatment principles:
- Lifestyle changes (proper diet, exercise regime) are the
basic and core problem in treatment. The time to evaluate the
effectiveness of lifestyle changes is usually 2-3 months.
- Medications when needed.
- Treatment goals based on tests and assessments of
patient risks, in which LDL-C is recommended as the first
treatment target. TC is the target for treatment if there are no
other tests, while non-HDL-C or ApoB is the second target.
* Drugs for dyslipidemia treatment according to
modern medicine:
Based on the mechanism of action, drugs for dyslipidemia
treatment are divided into 2 main groups as follows:
- The group that reduces lipid synthesis: Statins, fibric acid
(Fibrat), and nicotinic acid (Niacin)
- The group that reduces absorption and increases lipid
elimination: bile acid sequestrants (Resin), cholesterol
absorption inhibitor (Ezetimib)
- In addition, some other drugs are used to treat
dyslipidemia, such as PCSK9 inhibitors, MTP inhibitors, CETP
inhibitors, and so on.
1.2. Dyslipidemia from traditional medicine point of
view.
1.2.1. Causes
In terms of traditional medicine, dyslipidemia is caused by

the following syndromes: irregular congentinal factors,
uncontrolled diet leading to spleen-stomach disharmony;
internal injury leading to liver-gallbladder damage; the


4
debilitation of the elderly leading to the debilitation of source qi
liver. When the function of the viscera and bowels is
disordered, it causes phlegm-dampness pattern.
1.2.2. Disease mechanism
The process of disease development consists of three
stages:
- The dysfunction of spleen and kidney affects the
metabolism of water and food substances causing
endogenous dampness turbidity pattern.
- The function of damaged viscera and bowels causes
endogenous phlegm-dampness turbidity and pattern of
depressed dampness transforming into heat. The symptom in
this stage is phlegm turbidity depressing fire.
- The development of the disease causes binding of
phlegm stasis, qi damage, yin damage, and yin-yang
disharmony which can lead to a more serious condition and
damage of the visceral organs, resulting in complications.
1.2.3. Clinical classification in terms of traditional
medicine
- Pattern of internal phlegm-dampness: Body obesity,
headache, chest pain, abdominal distention, nausea or
vomiting, dry mouth without being thirsty, thin limbs, heavy
limbs, pale swollen tongue, white greasy coating tongue,
slippery pulse.

- Pattern of Phlegm-heat and fu-organ excess
syndrome: Strong well-being, heavy head, irritability, red
face, red eyes, bitter mouth, tight chest, uncomfortable,
indigestion, constipation, pink red tongue, yellow greasy
coating tongue, slippery pulse.
- Pattern of Spreen-kidney yang deficiency: puffy face,
pain in back and knees, fear of cold, mental fatigue,
indigestion, sloppy stool, night urination, pale tongue, edge of
tongue with teeth, thick white fur tongue, slow sunken pulse.
- Pattern of liver-kidney yin deficiency: Pain in back and
knees, irritable people, head pain, dizziness, fatigue, tinnitus,
nightly sweats, dry and thirsty mouth, red tongue, thin fur
tongue, ripid fine pulse.
- Pattern of phlegm stagnation and blood statis: Fatty
body, heavy and numbness limbs, tight chest, headache,


5
dizziness, purple fur & spotted tongue, thick fur tongue, strinklike and slippery pulse or rough pulse.
- Pattern of liver depression and spleen deficiency:
Tight pain in the edge of ribs, unstable pain, headache,
dizziness, irritability, poor appetite, mental fatigue, sloppy
stool, menstrual disorders, pale tongue, thin fur tongue, strinklike pulse.
1.3. Overview of therapeutic remedy research.
According to folk experiences and documents on traditional
medicines, there are many medicinal herbs, remedies for
treatment of phlegm-dampness pattern, which are proved to
be clinically effective in adjusting dyslipidemia. Most remedies
have too many herbs or they derive from abroad, making it
difficult for patients to take drugs for the prolonged treatment.

HVT remedy is based on the theory of traditional medicine and
some studies of modern medicine on the effectiveness of each
herb. HVT remedy is made up of Vietnamese medicinal herbs,
so they are easy to find, easy to use and have been commonly
used by people:
+ Folium Nelumbinis nuciferae which acts on upbearing the
clear, dissipating stasis, moving water, removing phlegmdampness. There have been many domestic and foreign
studies published results that Folium Nelumbinis nuciferae has
the effects of lowering blood pressure, lowering blood sugar,
preventing obesity, adjusting dyslipidemia, so it has the effect
of preventing atherosclerosis.
+ Cleistocalyx operculatus Roxb is a drink that has been used
for many generations in Vietnam as well as in many countries
around the world. It has the effect of fortifying the spleen and
removing the excess. There have been a lot of studies on
Cleistocalyx operculatus Roxb which have found that it is not
only a healthy drink, but also significantly effective in many
activities:
anti-microbial,
anti-dyslipidemia,
lowering
atherosclerosis, lowering blood sugar, inhibiting the
development of cancer cells, improving congestion, soothing
gallbladder, releasing pain and anti-allergy.
+ Pericarpium Citri reticulatae perenne is effective in fortifying
spleen, regulating qi, and drying dampness to resolve phlegm.
Pericarpium Citri reticulatae perenne is widely used as
medicine and food in Vietnam and many countries in the

6
world. This herb has been presented in more than 10% of
Chinese pharmacopoeia formulations. There have been many
studies proving the effect of Pericarpium Citri reticulatae
perenne in treating dyslipidemia, reducing obesity and
preventing atherosclerosis.
The effects of HVT remedy are fortifying spleen, regulating
qi, drying dampness and resolving phlegm.
Chapter 2 . SUBJECTS AND METHODS OF RESEARCH
2.1. Research materials
2.1.1 Experimental drug: HVT remedy is produced in liquid
form with a ratio of 1:1 (g/ml), bottled with volume of 240ml
(Folium Nelumbinis nuciferae 90g, Cleistocalyx operculatus
Roxb 90g, Pericarpium Citri reticulatae perenne 60g) by
Department of Pharmacology, Military Institute of Traditional
Medicine, meeting the basic standards.
2.1.2 Control drug (Statin group): Experimental study:
Atorvastatin 10mg (Lipistad-Stada Vietnam). Clinical study:
Atorvastatin 10mg (Lipitor-Pfizer Pharmaceutic Germany).
2.2. Subjects of research
2.2.1. Research subjects on experimental study
- White Swiss mice, healthy, weighing 200 ± 20g, provided
by National Institute of Hygiene and Epidemiology.
- White Wistar rats, healthy, weighing 150 – 180g, provided
by Military Medical University.
2.2.2. Research subjects on clinical study
- Criteria for selecting patients
* In terms of modern medicine: the patients over 40 years
old, regardless of occupation and gender. The patients are
diagnosed with dyslipidemia when they have at least one of

the following four abnormal indicators:
TC > 5.2 mmol/l, TG > 2.3 mmol/l; LDL-C > 3.4 mmol/l; HDL-C
< 0.9 mmol/l.
* In terms of traditional medicine: the patients have
manifestations of phlegm-dampness such as obesity, pale
swollen tongue, white greasy coating tongue, dizziness, severe
head pain, numbness of limbs, fatigue, palpitations, insomnia,
slippery pulse or string-like pulse.
- Exclusion criteria: the patients with chronic diseases, liver


7
failure, renal failure, secondary dyslipidemia syndrome or
being taking drugs that affect blood lipid index; the patients
with hypersensitivity to the drug ingredients; the patients with
a history of myocardial infarction, brain stroke, pregnancy or
breastfeeding; the patients do not abide by the treatment
rules.
2.3. Research methods
2.3.1. Experimental research methods
- Acute toxicity study: According to the method of
Litchfield - Wilcoxon, the guidelines of the Vietnam Ministry of
Health, WHO and OECD, Swiss white mice are divided into
groups with 10 mice per group. Mice in each group were taken
HVT at the dose from the highest dose (non-lethal dose) to the
lowest dose (lethal dose). Mice were completely starved for 12
hours before taking HVT (but drunk freely). The mice were
monitored the death rate in the first 72 hours as well as their
general condition for 7 days after taking the drug (including
eating activity, mental activity, walking, climbing, excretion,

and so on). Died mice were operated to evaluate the injuries of
organs and then calculated LD50.
- Semi-chronic toxicity study: According to guidelines
of the Vietnam Ministry of Health, WHO and OECD, Wistar
white rats were divided into 3 groups with 10 rats per group.
Rats in control group were drunk distilled water 1ml/100g/day
while rats in experimental group 1 were taken low dose HVT of
11.2 g medicinal/kg (dose equivalent to the dose used on
humans), and rats in experimental group l2 were taken highdose HVT of 33.6g medicinal/kg (3 times higher than
experimental group 1). Rats were drunk solvents and
experimental drugs for 8 weeks, once a day in the morning.
After 8 weeks, the rats stopped taking the drugs and were
monitored for 2 weeks to evaluate the ability of rehalilitation (if
there was toxicity) or the causing ability of the slow toxicity of
the drug. Monitoring indicators before and during the study
include general condition, body weight, assessment of
hematopoietic function, assessment of liver and kidney
function and histopathology of liver and kidney of white rats.
- Studying the effect of HVT liquid on dyslipidemia
adjustment in the model of exogenous hyperlipidemia


8
Rats were divided into 5 groups. Each group had 10 rats
with the same male/female ratio. All these groups were given
medicine for 4 weeks as follows: Group 1 (biological control
group) was drunk distilled water 1ml/100g rat. Group 2 (model
group) was drunk cholesterol oil mixture 1ml/100g, then drunk
distilled water 1ml/100g after 2 hours. Group 3 (atorvastatin
group) was taken 1 ml/100g cholesterol oil mixture, then taken

atorvastatin 10 mg/kg after 2 hours. Group 4 (the 1st dosage of
HVT) was taken cholesterol oil mixture 1 ml/100g, then taken
HVT dose 11.2g/kg after 2 hours. Group 5 (the 2 nd dosage of
HVT) was taken cholesterol oil mixture 1 ml/100g, then taken
HVT dose 33.6g / kg after 2 hours.
The rats in all groups were weighed before and after one,
two and four weeks of the experiment. Then, they were taken
blood at their tails to do quantative tests of blood lipid indices:
TC, TG, HDL-C. The non-HDL-cholesterol index is calculated by
the formula: non-HDL-C = TC - (HDL-C).
- Model of endogenous hyperlipidemia:
The model of endogenous hyperlipidemia was adjusted
by Poloxamer 407 (P-407) according to Millar et al. The 50
white mice were divided into 5 groups: Group 1 (white control
group), Group 2 (model group), Group 3 (atorvastatin group),
Group 4 (HVT dose of 19.2g/kg), and Group 5 (HVT dose of
96g/kg). The mice were taken HVT, atorvastatin or distilled
water within 7 days constantly before taking intraperitoneal
injection P-407. After taking P-407 injection, the mice were
completely starved and drunk freely. Within 24 hours after
receiving P-407, the mice were taken blood at the carotid
arteries to do quantative tests of TC, TG, HDL-C, non- HDL- C.
The index of non-HDL-choleserol was calculated based on
formula Non-HDL-C = (TC) – (HDL-C).
2.3.2. Clinical research methods
- Prospective study conducted randomized, opened clinical
trial with control group, result comparison between pretreatment and post-treatment. The 120 patients compliance
with diet and exercise were devided into 2 groups as follows:
+ Group 1: The experimental group consisted of 60 patients
who used HVT liquid with 80gam/day, 2 times per day, 30

minutes after lunch and dinner.


9
+ Group 2: The control group consisted of 60 patients who
used atorvastatin 10mg, 1 tablet per day, 1 hour after dinner.
The patients were examined, taken case history,
performed hematologic tests and taken biochemical blood
tests before treatment (D0) and after 30 days of treatment
(D30). The patients were taken venous blood in the morning
(without eating anything at least 12 hours before the previous
meal). Blood biochemical tests were done afterward (TC, TG,
LDL-C, HDL-C).
Evaluation of treatment results in terms of
modern medicine:
+ We evaluated the change of each lipid index at the
times of D0 and D30, and compared the results of each group
and between 2 groups as well.
+ The effectiveness of HVT in the patients with
dyslipidemia was evaluated by criteria as follows:
• Excellent: All blood lipid components returned to
normal limits, or one of them achieved the following changes:
TC decreased by ≥ 20%, TG decreased by ≥ 40%, HDL- C
increased by ≥ 0.26mmol/l, TC-HDL-C/HDL-C (non-HDL-C/HDLC) decreased by ≥ 20%.
• Good: when achieving one of the following criteria: TC
reduced by 10% - 20%, TG decreased by 20% - 40%, HDL-C
increased by 0.104mmol/l - 0.26 mmol/l, TC-HDL-C/HDL-C (nonHDL-C/HDL-C) decreased by 10% - 20%.
• Ineffective: Blood lipid components did not achieve the
above changes.
Evaluation of treatment results in terms of

traditional medicine
+ The effectiveness of HVT was evaluated through the
methods of inspection, inquiry and palpation.
+ The effectiveness of HVT in the patients with
dyslipidemia was evaluated by criteria as follows:
• Excellent: There were significant improvements in
clinical symptoms with the total decreased score ≥ 70%.
• Good: There were small changes in clinical symptoms
with the total decreased score ≥ 30%.
• Ineffective: The total decreased score of clinical
symptoms was less than 30%.
Evaluation of unwanted effects of HVT


10
Clinical evaluation: Some symptoms such as abdominal
pain, nausea, rash, diarrhea, muscle pain, etc were evaluated.
Sub-clinical evaluation: The results of hematological test
and blood biochemical tests before and after treatment were
used as good indicators to assess.
Data analysis: The data was collected and analyzed by
the method and Biomedical statistical algorithm of STATA 14.0.
Research Ethics: The study was approved by the
Medical Council of Traditional Medicine Hospital, Ministry of
Public Security. The patients, who voluntarily participated in
the study, were performed tests and given treatment without
paying any fee. All information was kept confidential and only
general results were allowed to publish.
Chapter 3. RESEARCH RESULTS
3.1. Acute toxicity and sub-chronic toxicity of HVT

liquid drug
3.1.1. Acute toxicity
White mice were taken HVT by increasing doses from
the lowest to the highest dose with 600.0g medicinal/kg/24
hours (up to 31.25 times higher than the equivalent dose on
the human) but there were no abnormal signs within 7 day
follow-up and no mice died within 72 hours after taking the
sample drug. Therefore, the acute toxicity of LD50 of HVT has
not been determined by oral administration.
3.1.2. Sub-chronic toxicity
* General condition: During the experiment, the rats in
all 3 groups were operating normally with bright eyes, smooth
hair, good eating, dry feces. There were no particular
manifestations in these three groups of rats during the study
period.
* Changes in body weight of rats: At 4-8 weeks after
taking the sample drugs and 2 weeks after stopping the drugs,
the weight of rats in all 3 groups were documented to be
increased compared to the weight before the study. There was
no difference in the level of weight gain of the rats between
the control group and the experimental group (p> 0.05).
* The effect of HVT on the mice’s blood lipid index


11
Table 3.1. The effect of HVT on the number of rat’s red
blood cells

Time
Before taking

drugs
4 weeks
after taking
drugs
8 weeks
after taking
drugs
2 weeks
after
stopping
drugs
P(before-after)

The number of red blood cells
(T/l)
The control
The group
The group
group (n=10)
1 (n=10)
2 (n=10)
(1)
(2)
(3)
8,49 ±
8,27 ±
8,32 ±
0,56
0,34
0,55

p2-1, p31, p3-2
>0,0
5

8,54 ±
0,93

8,59 ±
0,45

8,44 ±
0,72

>0,0
5

8,92 ±
0,68

8,72±
0,73

8,63 ±
0,58

>0,0
5

8,70 ±

0,69

8,48 ±
1,41

8,82 ±
0,68

>0,0
5

> 0,05

> 0,05

> 0,05

Table 3.2. The effect of HVT on the concentration of
hemoglobin in rat blood

Times
Before taking
drugs
4 weeks
after taking
drugs
8 weeks
after taking
drugs
2 weeks

after

The concentration of
hemoglobin (g/dl )
The control
The group
The group 1
group
2 (n=10)
(n=10) (2)
(n=10) (1)
(3)
15,32 ±
15,37 ±
15,19 ±
0,65
0,76
0,83

p2-1, p31, p3-2
>0,05

14,46 ±
1,25

15,30 ±
0,74

15,12 ±
0,96

>0,05

15,43 ±
0,83

15,38 ±
1,13

14,40 ±
0,98

>0,05

16,48 ±
0,97

16,27 ±
1,00

16,37 ±
2,62

>0,05


12

Times
stopping

drugs
P(before-after)

The concentration of
hemoglobin (g/dl )
The control
The group
The group 1
group
2 (n=10)
(n=10) (2)
(n=10) (1)
(3)
> 0,05

> 0,05

p2-1, p31, p3-2

> 0,05

Table 3.3. The effect of HVT on the number of rat’s
white blood cells
Time
Before taking
drugs
4 weeks
after taking
drugs
8 weeks

after taking
drugs
2 weeks
after
stopping
drugs
P(before-after)

The number of white blood cells
(G/l)
The control
The group
The group 2
group (n=10)
1 (n=10)
(n=10) (3)
(1)
(2)
5,12 ±
4,53 ±
4,73 ±
1,48
0,80
1,57

p2-1,
p3-1,
p3-2
>0,
05

5,69 ±
1,41

4,96±
1,22

5,33 ±
1,52

>0,
05

5,23 ±
1,30

4,65 ±
1,14

4,90 ±
1,40

>0,
05

5,67 ±
0,82

4,90 ±
0,62

5,57 ±
1,50

>0,
05

> 0,05

> 0,05

> 0,05

Table 3.4. The effect of HVT on the number of
thrombocytes in rat blood
Times
Before taking
drugs

The number of thrombocytes
(G/l)
The control
The group
The group 2
group (n=10)
1 (n=10)
(n=10) (3)
(1)
(2)
478,3 ±

483,5 ±
472,5 ± 66,9
46,6
131,2

p2-1,
p3-1,
p3-2
>0,0
5


13

Times
4 weeks
after taking
drugs
8 weeks
after taking
drugs
2 weeks
after
stopping
drugs
P(before-after)

The number of thrombocytes
(G/l)
The control

The group
The group 2
group (n=10)
1 (n=10)
(n=10) (3)
(1)
(2)

p2-1,
p3-1,
p3-2

418,5 ± 89,2

445,5 ±
66,6

449,1 ±
104,9

>0,0
5

428,5 ± 36,1

439,2 ±
69,3

443,1 ±
85,9

>0,0
5

458,2 ± 11,9

433,3 ±
41,1

453,6 ±
118,1

>0,0
5

> 0,05

> 0,05

> 0,05

After 4 - 8 weeks of taking HVT and after 2 weeks of
stopping HVT, the number of red bloods cells, white blood cells,
thrombocytes and the hemoglobin concentration in both of the
experimental group 1 (low HVT doses) and the experimental
group 2 (high HVT doses) did not change in comparision to the
time before taking the sample drugs and to the control group
(p> 0.05).
* Evaluation of liver function
Table 3.5. The effect of HVT on the activity of AST (GOT)

in rat blood
Times
Before
taking drugs
4 weeks
after taking
drugs
8 weeks
after taking

The activity of AST (UI/l)
The control
The group 1 The group 2
group
(n=10) (2)
(n=10) (3)
(n=10) (1)
105,60 ±
108,90 ±
112,90 ±
9,58
14,00
14,32

p2-1,
p3-1,
p3-2

>0,0
5

119,60 ±
22,74

117,50 ±
17,53

118,90 ±
23,65

>0,0
5

113,80 ±
18,27

112,60 ±
14,50

108,70 ±
11,21

>0,0
5


14
drugs
2 weeks
after

stopping
drugs
P(before-after)

111,00 ±
14,20

103,14 ±
5,96

108,14 ±
19,78

> 0,05

> 0,05

> 0,05

>0,0
5

Table 3.6. The effect of HVT on the activity of ALT (GPT)
in rat blood
Times
Before
taking drugs
4 weeks
after taking
drugs

8 weeks
after taking
drugs
2 weeks
after
stopping
drugs
P(before-after)

The activity of ALT (UI/l)
The control
The group 1 The group 2
group
(n=10) (2)
(n=10) (3)
(n=10) (1)
57,30 ±
61,70 ±
61,60 ±
5,33
4,76
7,26

p2-1,
p3-1,
p3-2

>0,0
5

62,80 ±
14,47

60,20 ±
9,10

60,10 ±
9,26

>0,0
5

59,30 ±
7,10

59,20 ±
6,78

59,90 ±
6,10

>0,0
5

57,00 ±
5,44

56,71 ±
4,72

52,71 ±
9,62

>0,0
5

> 0,05

> 0,05

> 0,05

After 4 - 8 weeks of taking HVT and after 2 weeks of
stopping HVT, there was not any statistically significant
difference in the activities of AST and ALT in rat blood in both of
the experimental group 1 (low HVT dose) and the experimental
group 2 (high HVT dose), in comparison to the control group and
between the two times before and after taking the sample
drugs (p> 0.05).
* Evaluation of kidney function
Table 3.7. The effect of HVT on the concentration of
creatinine in rat blood


15

Times
Before taking
drugs
4 weeks after

taking drugs
8 weeks after
taking drugs
2 weeks after
stopping drugs
P(before-after)

The concentration of
Creatinine (mg/dl)
The control
The group The group
group
1 (n=10)
2 (n=10)
(n=10) (1)
(2)
(3)
1,06 ±
1,07 ±
1,06 ± 0,05
0,05
0,05
1,05 ±
1,03 ±
1,04 ± 0,05
0,05
0,05
1,05 ±
1,05 ±
1,05 ± 0,05

0,05
0,05
1,04 ±
1,04 ±
1,03 ± 0,08
0,05
0,10
> 0,05
> 0,05
> 0,05

p2-1, p31, p3-2

>0,0
5
>0,0
5
>0,0
5
>0,0
5

After 4 - 8 weeks of taking HVT and after 2 weeks of
stopping HVT, there was not any statistically significant
difference in the concentration of creatinine in both of the
experimental group 1 (low HVT dose) and the experimental
group 2 (high HVT dose), in comparison to the control group and
between the two times before and after taking the sample
drugs (p> 0.05).
3.2. The pharmacological effect of HVT in experiment

study
Table 3.8. The lipid index at the time after 4 weeks of
experiment in exogenous model (n=10)
Groups
Group 1 :
The biological
control group
Group 2 :
The model
group
Group 3:
Atorvastatin
group
Group 4: The

The concentration of blood lipid indices
Non-HDL TC
TG
HDL - C
C
2,29 ± 0,07

0,77 ±
0,02

0,81 ±
0,03

1,48 ± 0,08

4,65 ±
0,21***

0,93 ±
0,04**

1,76 ±
0,05***

2,89 ±
0,2***

3,33 ±
0,18***

0,85 ±
0,04

1,58±
0,09

1,75 ±
0,18**

2,92 ±

0,87 ±

1,49 ±

1,43 ±


16
1st dose of
HVT (11,2
g/kg)
Group 5 :
The 2nd dose
of HVT (33,6
g/kg)

0,09***

0,03

0,05**

0,11***

2,96 ±
0,11***

0,89 ±
0,03

1,55 ±
0,04**

1,41 ±

0,11***

Notes: The differences in comparison to the model group: *: p <
0,05; **: p<0,01; ***: p < 0,001.
There was a significant decrease in the concentration of
TC and non-HDL-C in in both doses of 11.2 g/kg and 33.6 g/kg,
in comparison to the model group (p <0.001). The effect of 2
doses of HVT was similar and equivalent to atorvastatin 10 mg /
kg (p> 0.05). There was no statistically significant difference in
the concentration of blood lipid index between the atorvastatin
group and two HVT doses at the time after 4 weeks of
experiment (p> 0.05).
Table 3.9. Blood lipid index after the experiment in
endogenous model (n = 10)
Groups

TC

TG

HDL-C

Group 2:
The
model
group

Non-HDLC

6,31 ±

1,60

9,01 ±
2,36

2,11 ± 0,17

4,20 ±
1,68

Group 3:
Atorvastat
in group

4,15 ±
0,93**
(↓
34,2%)

8,16 ±
2,65
(↓ 9,4%)

2,02 ± 0,12

Group 4:
HVT
19,2g/kg

5,89 ±

1,37
(↓ 6,7%)

8,69 ±
3,90
(↓ 3,6%)

2,78 ±
0,38***
(↑ 31,8%)

Group
5:
HVT
96,0g/kg

4,91 ±
0,51 *
(↓
22,2%)

5,74 ±
1,79**
(↓
36,3%)

2,88 ±
0,48***
(↑ 36,5%)

2,13 ±
0,94**
(↓
49,3%)
3,12 ±
1,37
(↓
25,7%)
2,03 ±
0,89**
(↓
51,7%)


17
Notes: The difference in comparison to the model group: *: p <
0,05; **: p<0,01; ***: p < 0,001.
There was a significant increase in the concentration of
HDL-C in the group of HVT dose of 19.2g/kg in comparison to
the model group, with statistical significance (p <0.001). This
effect was stronger than the atorvastatin group of 100 mg/kg.
There was a significant decrease in the indices of TC, TG,
and non-HDL-C in the HVT dose of 96.0 g/kg in comparison to
the model group (p<0,05, p < 0,01 and p < 0,01). The HVT
made a significant increase in the concentration of HDL-C in
comparison to the model group. This effect was stronger than
the atorvastatin group of 100 mg /kg (p<0,001).
3.3. The effect of HVT in the patients with
dyslipidemia
caused

by
internal
phlegmdampness
3.3.1. The effect of HVT through sub-clinical indices
Table 3.10. The changes of the sub-clinical indices
after treatment
Times
Indices
HVT
TC

Atorvastatin
p
HVT

TG

HDLC

Atorvastatin
p
HVT
Atorvastatin

D0

D30

6,22 ±
0,48

6,37 ±
0,72
> 0,05
2,45 ±
1,13
2,57±1,25

5,24 ±
0,48
5,20 ±
0,76
> 0,05
1,94 ±
0,60
2,13±0,9
5
> 0,05
1,37 ±
0,30
1,48 ±
0,26

> 0,05
1,17 ±
0,32
1,28 ±
0,34

Change
s

↓15,8%
↓18,4%

↓20,8%
↓17,1%

↑17,1%
↑15,6%

p
<0,0
01
<0,0
01
<0,0
01
<0,0
1
<0,0
01
<0,0
01


18
p
HVT
LDLC

Atorvastatin

p
HVT

NonHDLC

Atorvastatin
p

> 0,05
3,93 ±
0,64
3,91 ±
0,74
> 0,05
5,05 ±
0,51
5,09 ±
0,66
> 0,05

> 0,05
2,99 ±
0,51
2,88 ±
0,80
> 0,05
3,87 ±
0,52
3,72 ±
0,73

> 0,05

↓23,9%
↓26,3%

↓23,4%
↓26,9%

<0,0
01
<0,0
01
<0,0
01
<0,0
01

After 30 days of treatment, the concentration of TC, TG,
LDL-C, non-HDL-C decreased significantly in both groups
compared to the time before treatment. The difference at the
time before and after treatment of 2 groups was statistically
significant. The difference in the degree of reduction in the subclinical indices between the two groups was not statistically
significant (p> 0.05).
The concentration of HDL-C increased markedly in both
groups compared to the time before treatment. The difference
at the time before and after treatment of 2 groups was
statistically significant (p <0.001). The difference in the
increasing level of HDL-C between the two groups was not
statistically significant (p> 0.05).
* Evaluation of treatment results in terms of

modern medicine
Diagram 3.1. Evaluation of the treatment effect in
terms of modern medicine
In terms of modern medicine, the treatment effect in HVT
group was shown as follows: excellent percentage was 51.7%,
good percentage was 43.3% and ineffective percentage was 5%.
The Atorvastatin group has achieved excellent result with 45.0%,


19
good result with 43.3%, and ineffective result with 11.7%. The
difference between the two groups was not statistically significant
(p> 0.05).
Table 3.11. The changes in risk assessment indices of
atherosclerosis after treatment
Times
Change
D0
D30
p
Indices
s
HVT

AI

Atorvastatin
p
HVT

CRI

Atorvastatin
p
HVT

AIP

Atorvastatin
p

4,62 ±
1,30
4,22 ±
1,11
> 0,05
5,62 ±
1,30
5,22 ±
1,11
> 0,05
0,66 ±
0,11
0,62 ±
0,10
> 0,05

2,98 ±
0,85
2,59 ±

0,69
> 0,05
3,98 ±
0,85
3,59 ±
0,69
> 0,05
0,22 ±
0,24
0,19 ±
0,26
> 0,05

↓35,50%
↓38,6%

↓29,2%
↓31,2%

↓67,1%
↓69,3%

<0,00
1
<0,00
1
<0,00
1
<0,0
01

<0,00
1
<0,00
1

After 30 days of treatment, the indices of AI (TC-HDL-C /
HDL-C), CRI (TC / HDL-C) and AIP (log (TG / HDL-C)) has been
decreased markedly in both group compared to the time before
treatment. The difference at the time before and after treatment
of 2 groups was statistically significant (p <0.001). The
difference in decreasing level of AI, CRI and AIP indices of the
two groups was not statistically significant (p> 0.05).
3.3.2. The effect of HVT on some clinical indicators
Table 3.12. The changes of clinical symptoms in
terms of inspection
Groups

HVT

Atorvastati
n

Total


20
Inspection

Fatty body

D0
(n %)
16
(26,7)

Thick &
41
teeth-marked
(68,3)
tongue
White & slimy
39
fur tongue
(65,0)

D30
(n %)
12
(20,0
)
12
(20,0
)
15
(25,0
)

D0
(n %)
11

(18,3
)
46
(76,7
)
38
(63,3
)

D30
D0
D30
(n %) (n %) (n %)
10
27
22
(16,7
(22,5) (18,3)
)
24
87
36
(40,0
(72,5) (30,0)
)
11
77
26
(18,3
(64,2) (21,7)

)

After treatment, the number of patients with fatty
condition was decreased by 22 patients (18.3%) compared to
27 (22.5%) before treatment. The number of patients with thick
& teeth-marked tongue was reduced by 36 (30.0%) after
treatment in comparison to 87 patients (72.5%) before
treatment. There was also a significant reduction in the number
of patients with white & slimy fur tongue before and after
treatment (77 patients (64.2%) compare to 26 patients
(21.7%)).
Table 3.13. The changes of pulse in terms of
traditional medicine
Groups

HVT
D0
D30
(n %)
(n %)

Atorvastatin
D0
D30
Pulpation
(n %)
(n %)
Moderate &
14
0 (0,0) 23 (38,3) 0 (0)

soggy pulse
(23,3)
Sunken
0 (0) 11 (18,3) 0 (0,0) 9 (15,0)
pulse
String-like
10
7 (11,7) 11 (18,3) 8 (13,3)
pulse
(16,7)
Slippery
21
13
10 (16,7) 23 (38,3)
pulse
(35,0)
(21,7)
Slippery
&
29
16
string-like
9 (15,0) 26 (43,4)
(48,3)
(26,7)
pulse

Total
D0
D30

(n %) (n %)
37
0 (0,0)
(30,8)
20
0 (0,0)
(16,7)
21
15
(17,5) (12,5)
44
23
(36,7) (19,2)
55
(45,8)

25
(20,8)


21
The number of patients with slippery & string-like pulse
in the HVT group was significantly decreased after treatment
(9 patients (15.0%) compared to 29 patients (48.3%)). The
number of patients with slippery & string-like pulse in the
atorvastatin group was also reduced after treatment (16
patients (26.7%) compared to 26 patients (43.4%)). There
were 21 patients with slippery pulse before treatment,
however, after receiving HVT treatment, the number of
patients with slippery pulse was also significantly reduced by

10 patients (16.7%). In the Atorvastatin group, there were 13
patients (21.7%) after treatment compared to 23 patients
(38.3%) before treatment.
Table 3.14. The changes of clinical symptoms in
terms of inquiry
Groups

D0 (n %)

Symptoms

Headach
e

No
pain

9
(15,0)

Mild

3 (5,0)

Modera
18
te
(30,0)
30
Severe

(50,0)
No
2 (3,3)
pain
Mild

Dizziness

HVT

3 (5,0)

Modera
26
te
(43,3)
29
Severe
(48,3)
Numbn
No
10
ess
in
pain
(16,7)
limbs
Mild
11
(18,3)

Atorvastatin
Total
D30 (n
D30 (n %) D0 (n %)
D0 (n %) D30 (n %)
%)
30
14
44
0 (0)
9 (7,5)
(50,0)
(23,3)
(36,7)
30
41
71
0 (0)
3 (2,5)
(50,0)
(68,3)
(59,2)
29
47
0 (0)
5(8,3)
5 (4,2)
(48,3)
(39,2)

31
61
0 (0)
0 (0)
0 (0)
(51,7)
(50,8)
27
13
40
0 (0)
2 (1,7)
(45,0)
(21,7)
(33,3)
33
43
76
4 (6,7)
7 (5,8)
(55,0)
(71,7)
(63,3)
40
66
0 (0)
4 (6,7)
4 (3,3)
(66,7)
(55,0)

16
45
0 (0)
0 (0)
0 (0)
(26,7)
(37,5)
22
11
18(15,
33
8 (13,3)
(36,7)
(18,3)
0)
(27,5)
38
11
47
22
85
(63,3)
(18,3)
(78,3) (18,3) (70,8)


22
Modera
22
te

(36,7)
17
Severe
(28,3)
No
1 (1,7)
pain
8
Mild
(13,3)
Insomnia
Modera
45
te
(75,0)
6
Severe
(10,0)

0 (0)
0 (0)
12
(20,0)
43
(71,7)
5 (8,3)
0 (0)

31
(51,7)

10
(16,7)
10
(16,7)
10
(16,7)
39
(65,0)
1 (1,7)

2 (3,3)
0 (0)
19
(31,7)
38
(63,3)
3 (5,0)
0 (0)

53
(44,2)
27
(22,5)
11
(9,2)
18
(15,0)
84
(70,0)
7 (5,8)

2 (1,7)
0 (0)
31
(25,8)
81
(67,5)
8 (6,7)
0 (0)

In both groups, some symptoms caused by phlegmdampness such as severe headaches, dizziness, numbness in
limbs, insomnia have been significantly improved after
treatment compared to them before treatment. The
difference before and after treatment in both groups was
statistically significant (p <0.05).
* Evaluation of the treatment results in terms of
traditional medicine
Diagram 3.2. Evaluation of the treatment effect in
terms of traditional medicine
The effectiveness of treatment according to traditional
medicine in the HVT group achieved excellent results with
38.3% and good results with 61.7%. There was not any patient
who was ineffective to the treatment. The Atorvastatin group
achieved excellent results with 10%, good results with 85%, and
ineffective results with 5%. The difference between the two
groups was statistically significant (p <0.05).
3.3.3. Evaluation of unwanted effects
No unwanted effects have been found in the clinical study.
Chapter 4. DISCUSSIONS
4.1. Acute toxicity and semi-chronic toxicity of HVT

liquid drug


23
4.1.1. Acute toxicity
White mice were taken HVT with the increasing dose from
360.0g/kg/24 hours to 600.0g/kg/24 hours (31.25 times higher
than the dose used in human). After taking the sample drug, no
mice died and no abnormal manifestation were observed in
mice. Therefore, acute toxicity and LD50 of HVT have not been
determined.
4.1.2. Semi-toxic toxicity
After 8 weeks of taking the medicine and after 2 weeks of
stopping the drug on rats, the results showed that HVT with
dose of 11.2g/kg and 33.6g/kg did not affect the development
of rats, and there were no changes observed in the indicators of
hematology, blood biochemistry and general images,
histopathology of liver and kidney in rats. Therefore, HVT has
not shown a semi-chronic toxicity of white rats by oral
administration.
4.2. The pharmacological effects of HVT in the
experimental study
4.2.2. The model of exogenous dyslipidemia
Both HVT doses have showed a significant lipid-lowering
effect after 4 weeks of study. This was expressed by a
statistically significant reduction in the concentration of TC and
non-HDL-C compared to the model group (p <0.001). This effect
was equivalent to the atorvastatin group at 10 mg/kg/day (p>
0.05). Lipid-lowering effects of low-dose HVT were not
statistically significantly different from high-dose HVT and this

was equivalent to the atorvastatin group at 10mg/kg/day.
Therefore, HVT has shown the good effect of adjusting
dyslipidemia on the model of exogenous dyslipidemia. This
effect did not depend on whichever high-dose or low-dose HVT
(p> 0.05).
4.2.2. The model of endogenous dyslipidemia
The indices of TC, TG, non-HDL-C in the mice’s serum of
the group taking HVT dose of 19.2g/kg/day tended to decrease
compared to the model group. There was a statistically


24
significant reduction in the group taking HVT dose of
96.0g/kg/day. The TC-lowering effect of high-dose HVT was
lower than that of the atorvastatin at a dose of 100 mg/kg
(22.2% compared to 34.2%), however the TG-lowering effect
was significantly higher than that of the atorvastatin (36.3%
compared to 9.4%). In this study, HVT at both doses has
showed a significant increase in HDL-C, better than the
atorvastatin group (p <0.001). From these results, HVT has
shown the good effect of adjusting dyslipidemia on the model
endogenous dyslipidemia by P-407. This effect did not depend
on whichever HVT high-dose or low-dose.
Thus, HVT has shown the effect of adjusting dyslipidemia
on both models of endogenous and exogenous dyslipidemia.
Based on published studies, there are quite a number of lipidlowering mechanisms that can partly explain the effect of
dyslipidemia adjustment of the HVT remedy.
The polymethoxyflavon components in Pericarpium Citri
reticulatae perenne have been shown to inhibit the ability of
pancreatic lipase, in which nobiletine has been proved to be the

strongest
lipase
inhibitor.
In
the
hamsters
with
hypercholesterolemia by diet, polymethoxylated flavones
(mainly tangeretin) and glucosid flavanone (hesperidin and
naringin) in Pericarpium Citri reticulatae perenne have been
shown to decrease the concentration of VLDL and LDL, or tend
to reduce triacylglycerol in serum.
The lipid-lowering effect of Folium Nelumbinis nuciferae
has been documented in several studies. The study by Lee et al.
(2010) showed that rabbits fed a high-fat diet supplemented
with the liquid extract from Folium Nelumbinis nuciferae at a
ratio of 0.5% and 1% had a significant decrease in the
concentration of TG (30% and 46.6%) and LDL (45.4 and
45.9%) compared to the rabbits that only fed a fatty diet.
Cleistocalyx operculatus Roxb is often commonly used to
be one of the Vietnamese tea drinking. In vitro, the extract from
Cleistocalyx operculatus Roxb with the high content of phenol


25
and flavonoids has showed to be the most effective inhibitor on
lipase when compared to the extract from green tea and guava
leaves. Besides the effect of blood glucose reduction, the
extract from Cleistocalyx operculatus Roxb also showed the
significant reduction in the concentration of TC and TG in serum

when the rats with treptozotocin-induced diabetes were orally
administered the sample for 8 weeks (500 mg/kg day).
Catechin is also a rich ingredient in Cleistocalyx operculatus
Roxb, therefore, the lipid-lowering mechanism of Cleistocalyx
operculatus Roxb may also be related to the change of gene
manifestations caused by lipid metabolism.
4.3. Evaluation of the effect of HVT in the patients with
dyslipidemia caused by internal phlegm-dampness
4.3.1. The effect of HVT through some clinical indicators
* The change of TC concentration
After 30 days of treatment, the concentration of TC was
significantly reduced in both groups compared to it before
treatment. There was a statistical significant difference between
the two groups before and after treatment (p <0.001). The
difference in the TC reduction between the two groups was not
statistically significant (p> 0.05). According to Goode GK et al.,
who analyzed 35 large-scale studies on 77.257 patients who
were monitored from 2 to 12 years, has concluded that if there
is the reduction in TC by 20%, the mortality will reduce by
18.1% in general and 24.1% in the patients with coronary artery
disease. Thus, how to reduce the concentration of TC is one of
the top goals in dyslipidemia treatment.
The decrease in TC concentration of HVT is also relevant
to the results of TC lowering effect on the model of exogenous
hyperlipidemia in our study.
* The change of TG concentration
The concentration of blood TG is a key indicator to explore
the body's lipid balance and contribute to reflecting the risks of
atherosclerosis.
After 30 days of treatment, the concentration of TG was