Traditional Chinese medicine in treatment of hyperlipidaemia

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ETHNO PHARMACOIDGY ELSEVIE R

Journal of Ethnopharmacology46 (1995) 125-129

Short communication

Traditional Chinese medicine in treatment of hyperlipidaemia B i r g i t t e la C o u r *a, P e r M O l g a a r d a, Z h a o Y i b aDepartment of Pharmacognosy, 2 Universitetsparken, 2100 Copenhagen, Denmark bGuangxi College of Traditional Chinese Medicine. 21 Ming Xiu Road, Nanning, 530001 Guangxi, P.R China

Received 5 January 1993; revision received 17 September 1994; accepted 30 January 1995

Abstract

In the search of new products for treatment of hyperlipidaemia with a low frequency of side effects a decoction of Crataegus cuneata, Nelumbo nucifera and Gynostemma pentaphylla has been tested. A reduction of triglyceride and cholesterol was seen. Keywords: Hyperlipidaemia reduction; Decoction; Traditional Chinese medicine

1. Introduction

Recently, the industrial world has seen an increase in problems due to hyperlipidaemia and there is, consequently, an increasing demand for a medical treatment, with a low frequency of side effects for this problem. Traditional Chinese medicine has used herbs to treat hyperlipedemia, and it has been shown that Nelumbo nucifera Gaertn. (Nymphaeaceae), reduces the total free cholesterol and phospholipid counts although it does not lower the serum cholesterol (Onishe et al., 1984). Crataegus cuneata Sieb. et Zucc. (Crataegeae) has been used in a decoction (Sakai et al., 1989), and was shown to increase the nicotinamide ade* Corresponding author.

nine dinucleotide (NAD) phosphate. C. cuneata also inhibits a drop in the activity of adenine dinucleotide (ADH) reduced form in the liver, which seems to improve ethanol catabolism. C. cuneata has also proved valuable as a general protection agent for liver metabolism (Sakai et al,. 1989). In a study made at the Third Affiliated Hospital of Beijing University of Medical Science Gynostemma pentaphylla (Thunb.) Mark. (Cucurbitaceae) has been shown to possess a strengthening and lipid-reducing effect (Chinese Academy of Medical Sciences et al., 1990). The present project was carried out at the College of Traditional Chinese Medicine, Nanning, Guangxi, P.R. China, according to Chinese medical tradition on a mixture of plants. In the investi-

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B. la Cour et aL /Journal of Ethnopharmacology 46 (1995) 125-129

gation to reduce serum lipid levels, the mixture used contained three plant species: C. cuneata, N. nucifera and G. pentaphylla. The selection of these plants was based on the tradition of Guangxi. The project was designed to evaluate the ability of the decoction to lower cholesterol and triglyceride in a short term experiment with rats and quails, using atromid as a standard treatment (Prior and Campbell, 1963). The experiment was carried out according to the method of (Wang Wei, 1984, 1990). Although atromid has very few side effects, it is not advised for patients with renal or liver problems or who suffer from gallstones. Likewise pregnant and nursing women should avoid atromid (Kristensen et al., 1990). It is important that there should be a safe plant product available which would be of benefit in such cases. 2. Methodology 2.1. Animals A 1-week study using 110 albino rats, (150-170 g) and 60 Japanese quails (90-110 g) was carried out. None of the animals were full-grown when the treatment was started. The rats and quails were kept in plastic boxes, one rat/box and two quails/box. They received oral injections of water/decoction/atromid every day except the day the experiment was terminated. 2.2. Diet The animals were given free access to a standard and high lipid diet, respectively, and water. The high lipid part of the basic diet was prepared with the following ingredients: cholesterol 2 g, pigs' fat 10 g, methyl thiouracil 0.2 g, carboxyl methyl cellulose 1 g and Aq. dest. Ad. 20 ml. Methyl thiouracil was added as an antithyroid hormone to inhibit the animals' natural reaction against hyperlipidaemia. In this way it was possible to shorten the experiments. 2.3. Decoction The decoction was prepared from the dried plants extracted with water, boiled for 1 h and stored in a cold location before use. The plants

were grown in the Botanical Garden belonging to the Guangxi College of Traditional Chinese Medicine. The composition, in accordance with tradition, was as follows: C. cuneata (fructus) 0.7 g, N. nucifera (folium) 0.7 g, G. pentaphylla (herba) 0.7 g and Aq. dest. Ad. 20 ml. The effect of the decoction was compared to that of atromid 0.07 g/kg per day. 2.4. Blood Blood samples from the rats were collected from the eye socket. Blood samples from the quails were collected from the neck vena jugularis interna and the arteriae carotis communis after decapitation. 2.5. Triglyceride Triglycerides were quantified as described by Dunsbach (1966). After removal of the glycerol containing fractions by N-heptan-isopropanol (2:3.5 v/v) and 0.04 N sulfuric acid from the serum, the glycerol, obtained by saponification of the triglycerides, was oxidized to formaldehyde with periodate. The resulting formaldehyde was converted into diacetyldihydrolutidine (Hantzsch reaction) with acetyl acetone and ammonium salt in acetic acid solution. Triolein was used for comparison. The yellow amyl alcoholic solution was quantified spectrophotometrically at 420 nm. 2.6. Cholesterol Cholesterol was measured by a direct coiorimetric method (Davidsohn and Henry, 1974); the serum was mixed with acetate. When the acetate reagent extract was mixed with sulfuric acidferrous sulfate reagent, it produced a purple color within 15 min with a maximum absorbance at 560 nm; it was stable for at least 1 h. It correlates well with analyses using the Abell technique and appears to be practically free from interference. 2. 7. Statistics The treatments were compared using the t-test. 3. Results and discussion The results show that the decoction of N. nucifera, C. cuneata and G. pentaphylla has a signi-

B. la Cour et al. /Journal of Ethnopharmacology 46 (1995; 125-129

127

Cholesterol/Triglyceride 3~, Choleslerol

F-O

250-

Triglyceride

150-

IIXI-

50-

Normal

Fal

Dcco 2g

i

Deco 4g

Deca~8g

Alromid

Average of 20 replicates with rats Fig. 1. Results for one week rat experiment. Triglycerideand cholesterol in blood serum (mg/100 ml) as response to treatment with decoction compared to control standard (normal) and high lipid diet (fat) and treatment with atromid. was not seen, but rather a negative correlation with a m a x i m u m effect at 2 g/kg per day both for cholesterol and triglyceride was observed. Using the t-test the significance levels were as follows:

ficant reducing effect on the serum triglyceride and cholesterol. The experiment with rats (Fig. l) shows a significant effect of the decoction. A simple positive correlation between dose and effect Cholesterol/Triglyceride 1200-

foliO-

8 fl 400 -

2QO-

Normal

Fal

Dc~) 2g

Dcco4g

Dcco8g

Alromid

Average of 10 replicates with quails Fig. 2. Results for one week quails experiment. Triglycerideand cholesterol in blood serum (mg/100 ml) as response to treatment with decoction compared to control standard (normal) and high lipid diet (fat) and treatment with atromid.

128

B. la Cour et al./ Journal of Ethnopharmacology 46 (1995) 125-129

(a) cholesterol: P < 0.001 for standard diet, 2 g/kg per day decoction and 0.07 g/kg per day atromid; P < 0.01 for 4 g/kg per day decoction; no significant difference for 8 g/kg per day; (b) triglyceride: P < 0.001 for standard diet and 0.07 g/kg per day atromid; P < 0.01 for 2 g/kg per day decoction and 4 g/kg per day decoction; P < 0.05 for 8 g/kg per day decoction. The t-test results indicate that the doses used were probably too high and this suggests the use of doses below 2 g/kg per day. For the quails the 1-week results (Fig. 2) produced similar results for the different doses of the decoction. Looking at the t-test for the 1-week treatment we found: (a) cholesterol: P < 0.001 for standard diet and 8 g/kg per day decoction; P < 0.01 for 2 g/kg per day decoction, 4 g/kg per day decoction and 0.07 g/kg per day atromid; (b) triglyceride: P < 0.001 for standard diet 2 g/kg per day decoction, 4 g/kg per day decoction and 8 g/kg per day decoction; P < 0.05 for 0.07 g/kg per day atromid. The results do not show any dose/effect relationship, but in any case do indicate an effect as good as with atromid. We are not aware of previous reports on the lipid reducing effect of decoctions of a mixture of these three plant species. When using the three species together it is difficult to determine whether one of the plants is responsible for a prime effect, or whether the effect is due to an interaction between the plants during the preparation of the decoction or due to interaction of the plants once they are ingested into the body. Such interaction could be synergistic and increase the combined effect, but in this case it appears to be antagonistic and hence reduces the resultant effect. This would explain the negative correlation of the dose/effect in the experiment with rats and also the lack of correlation in the experiment with quails. Looking at the effects together it is worth noting, that N. nucifera has a cholesterol and phospholipid reducing effect (Onishe et al,. 1984), C. cuneata has a protecting effect on the liver metabolism (Sakai et al,. 1989) and G. pentaphylla has a strengthening and lipid reducing effect (Chinese Academy of Medical Sciences et al., 1990).

Probably the results are influenced by all these effects which do not appear to act synergistically. However, a more precise description of the metabolic processes is beyond the aim of this investigation. In the experiments no side effects were observed in either rats or quails. Future investigation could throw light on questions of precise dose/effect relationship, metabolism, metabolic side effects and long term effects. The isolation and elucidation of active chemical constituents should be given a high priority in future investigations.

Acknowledgments This investigation was carried out by Birgitte la Cour during a study period at College of Traditional Chinese Medicine, Nanning, Guangxi, P.R. China, at the Department of Experimental Pharmacology. Birgitte la Cour gratefully acknowledges the help of the staff who made it possible to perform this investigation. The study was made possible due to grants from Director Ib Henriksens Foundation, a scholarship from Copenhagen Kommune, Borgervennen of 1788 and 'De videreg~ende uddannelsers pulje'.

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