Study On The Mechanism For The Hypolipidemic Effect Of Alismatis Rhizoma Decoction

Part One:Related Literature ReviewHyperlipidemia is a common chronic disease, the incidence of which is due to the disorder of body lipid metabolism, and the accumulation of a large number of related substances, causing fatty liver, diabetes, arterial atherosclerosis, hypertension and cardiovascular diseases, and threatening people’s lives and health. The clinical manifestations of hyperlipidemia is higher concentrations of serum TC and TG and lower concentrations of HDL-C.There are many Chinese medicine and compound for the treatment of hyperlipidemia, and have achieved good effects. Alismatis Rhizoma Decoction is a classical Traditional Chinese Medicine formula for treatment of vertigo with its long history of efficacious and successful clinical effect. AD is composed of two component Chinese herbal medicine which are Alismatis Rhizoma and Atractylodis Macrocephalae Rhizoma. Since vertigo is a symptom of hyperlipidemia, the present study aimed at evaluating the hypolipidemic effect of ARD in hypolipidemic mice induced by high fat diet and investigated the rationality of formula combination of AR and AMR. Compared with mice in control group, hypolipidemic mice in AR and AD groups displayed a reduction of following parameters:body weight, liver and serum total cholesterol, triglyceride concentration, liver coefficient, activities of serum aspartate aminotransferase AST and alanine aminotransferase ALT; whereas the concentrations of serum HDL-cholesterol were significantly elevated in both AR and AD groups. AR and AD treatments significantly down regulated the expressions of3-hydroxy-3-methylglutharyl-coenzyme A reductase HMG-CoA reductase and sterol regulatory element binding factor-2SREBF-2.hese findings clearly provided evidences that molecular mechanisms of the hypolipidemic effects of AD and AR in hypolipidemic mice are due to their suppression on biosynthesis of cholesterol in liver. Since no significantly hypolipidemic effect of AMR was observed in hypolipidemic mice, the more prominent effect of AD than that of AR indicated a synergistic effect of AR and AMR, and confirmed the rationality of AD formula. The Alismatis Rhizoma Decoction is a classic prescription from the Synopsis of the Golden Chamber written by medical saint Zhang Zhongjing in Eastern Han dynasty. The prescription were composed by Alisma and atractylodes. Alisma’s role focuses on Lishuixiaxing, banishing dampness effect; Atractylodes’s role focuses on strengthening spleen and making water effects. The prescription for the treatment of faint and dizziness earliest, and later also for the treatment of hyperlipidemia, is actually a further expansion of Alismatis Rhizoma Decoction in applications, and the purpose and mechanism of drug were basically the same. long-term clinical validation proved the prescription has a good effect on hyperlipidemia. However, the research literature of Alismatis Rhizoma Decoction’s mechanism in hypolipidemic is few, and the mechanism is not clear and was still in the early exploratory stage. It is commonly believed that Alismatis Rhizoma Decoction can affect the absorption of exogenous cholesterol and Alismatis Rhizoma Decoction can interfere the endogenous cholesterol catabolism, and the hypolipidemic effect of Alismatis Rhizoma Decoction may be related to improving the body’s antioxidant capacity and reducing the body’s oxidative stress. Some scholars believe that the mechanism of some traditional Chinese medicine hypolipidemic drug may also related to the protective effect of liver, spleen, kidney and regulating the organ balance which is helpful for the maintainance of body’s normal physiological function, and the mechanism may also related to the regulation of lipidmetabolic disorders. The research of Alismatis Rhizoma Decoction’s mechanism by molecular biology is in the exploration stage. The study of genes in the liver that regulate cholesterol synthesis and the key enzymes(such as Hmgcr, Srebf2Fasn, Cyp7al, etc.) that affect the synthesis and decomposition of cholestero is one of the major mechanism of lipid-lowering drugs. The study in key gene that regulating the lipemia may be fundamental to clarify the drug effect on lipid. To clarify the mechanism of Alismatis Rhizoma Decoction hypolipidemic effect not only can guide clinical medication safety, but also meaningful for the development of new effective lipid-lowering drug.Part two:Experimental researchExperiment one:The preliminary mechanism study of Alismatis Rhizoma Decoction in hypolip-idemicObjective:observe the Alismatis Rhizoma Decoctions’s effect of blood lipids in hyperlipidemic mice and study the preliminary hypolipidemic mechanism of Alismatis Rhizoma Decoction. Methods: use high fat diet to establish hyperlipidemia model, Sixty male Kunming mice were randomly divided into normal control, high-fat control, positive control group, Alismatis Rhizoma Decoction extract high dose group, medium dose group and low dose group, each group consisting of10animals. In addition to normal group which was fed with normal diet, other four groups were fed with high-fat diet for4weeks to establish the experimental hyperlipemia model. Then the normal control and high-fat control were given distilled water orally, positive control group was given simvastatin, others were given Alismatis Rhizoma Decoction extract high, medium and low dose, continuous administration for4w, determinate hyperlipidemia mice in serum TC, TG HDL-C, SOD, MDA, ALT, and AST and other biochemical parameters, to observe the pathological changes in tissues and organs, and the liver biopsy stained with Hematoxylin-eosin (HE) were observed under light microscope. Results:After4w administration, hyperlipidemic mice serum TC, TG was significantly lower, and HDL-C, SOD significantly increased, ALT, AST also significantly reduced. Conclusion:Alismatis Rhizoma Decoction extract have a good effect in hyperlipidemic mice especially the high dose group. Its lipid-lowering mechanism may be the protection of the liver, kidney, spleen and other organs, and the balance regulating of body metabolism, thus to improve the antioxidant capacity, and reduce oxidative stress.Experiment two:LC-MS/MS Fingerprinting of the Alismatis Rhizoma DecoctionObjective:To establish LC-MS/MS fingerprint of Alismatis Rhizoma Decoction extract, and standardized the active lowering lipid ingredients of Alismatis Rhizoma Decoction, lay the foundation of Alismatis Rhizoma Decoction’s lipid-lowering mechanism. Methods:use LC-MS/MS techniques, the Alismatis Rhizoma Decoction extract as samples, to establish the active ingredients of the soup hypolipidemic fingerprint. Results:establish fingerprint of the active ingredients of the Alismatis Rhizoma Decoction. Conclusion:The method to establish the Alismatis Rhizoma Decoction fingerprint is convenient, feasible, has effective control in lipid-lowering active ingredients of Alismatis Rhizoma Decoction, and provides an experimental basis for the study of Alismatis Rhizoma Decoction hypolipidemic effect and the mechanism of lipid-lowering effect.Experiment three:Study on the mechanism for the hypolipidemic effect of Alismatis Rhizoma DecoctionObjective:To explore the Alismatis Rhizoma Decoction hypolipidemic effect mechanism by the partly prescription and combinated prescription Methods:Study compound Alismatis Rhizoma Decoction extract, a single the Alisma water extract, Sixty male Kunming mice were randomly divided into normal control, high-fat control, positive control group, Alismatis Rhizoma Decoction extract group, Alisma water extract group and Atractylodes water extract group, each group consisting of10animals. In addition to normal group which was fed with normal diet, other four groups were fed with high-fat diet for4weeks to establish the experimental hyperlipemia model. Then the normal control and high-fat control were given distilled water orally, positive control group was given simvastatin, others were given Alismatis Rhizoma Decoction extract, Alisma water extract and Atractylodes water extrac, continuous administration for4w. Determinate hyperlipidemia mice in serum TC, TG HDL-C, SOD, MDA, ALT, and AST and other biochemical parameters, to observe the pathological changes in tissues and organs, and the liver biopsy stained with Hematoxylin-eosin (HE) were observed under light microscope. The Atractylodes water extract of blood lipids in hyperlipidemic mice, and using modern molecular biology techniques to study the effect of Alismatis Rhizoma Decoction in the expression of key genes (Hmgcr, Srebf2, Fasn, Cyp7al) that regulating lipid. Results:Alismatis Rhizoma Decoction extract group still have a good effect on lowering the lipid, and Alisma water extract group also had a weak effect in lowering blood pressure, while the Atractylodes water extract group had basically no effect on lowering lipid. Alismatis Rhizoma Decoction more could inhibit the expression of Hmgcr, Srebf2, Fasn in the liver of hyperlipidemic mice significantly, but had no significant effect on the expression of Cyp7al. Conclusion:Alismatis Rhizoma Decoction has a good effect on lowering hypolipidemic effect and its lipid-lowering effect may be related to the combination with Atractylodes, mechanism of action with the Alisma Tang can regulate cholesterol synthesis and metabolism gene expression related to Chak spilled soup inhibition Hmgcr, SrebfZ, Fasn expression, inhibition of cholesterol synthesis, thereby reducing serum TC TG, and other biochemical markers, to the hypolipidemic effect.