Studies On Chemical Components Analyze Of Huanglian Jiedu Decoction And The Effect Of Anti-hyperlipidemia

Huanglian jiedu decoction, a classical Chinese traditional prescription which could clear away heat and toxic material, is composed of Coptis chinensis Franch,Scutellaria baicalensis Georgi,Phellodendron amurense Rupr and Gardenia jasminoides Ellis. Advanced researches about the chemical components in these four traditional drugs have been made. However, because of the complex of the components of huanglian jiedu decoction, there were few of study concerning the integrated prescription. Only some researches focus on the main compounds in huanglian jiedu decoction have been made, such as berberine, baicalin, wogonin and gardenside, less study on the components absorbed in serum. Therefore, with a spirit of that a component which is absorbed in serum might be the genuine activity component, we focus on the follow two aspects, one is the chemical components analysis of huanglian jiedu decoction, the other is detecting the serum of body after administrated huanglian jiedu decoction.First, using HPLC as a detection method, we have surveyed the concentration of berberine and baicalin in huanglian jiedu decoction and its different combinations of these four drugs, afterwards we detected the component discrepancies between huanglian jiedu decoction before and after administrated using LC-MS. In this test, the concentration of berberine in Coptis chinensis Franch is 3.04%, the ratio of baicalin in Scutellaria baicalensis Georgi stands at 6.94%. However, in huanglian jiedu decoction, the concentration of berberine is only 0.17%, as to baicalin, the concentration reached to 10.83%. Through these results, we noticed that the components of huanglian jiedu decoction are not just a simple amount of the components of these four drugs. In order to explore the discrepancy of chemical components in the whole prescription and in each kind of drug, we made different combination of these four drugs, after obtained different LC-MS results of each sample, we analyzed all of 84 peaks in chromatograms of huanglian jiedu decoction, compared with the chromatograms of other combinations and seek the original source of these components. We got serum of rats after administrated huanglian jiedu decoction at the same time and compared its chromatogram with chromatograms of the serum containing other different combinations and of six standard materials, berberine, baicalin, balcalein, wogonin, wogonoside and geniposide. Among this datas, 14 compounds from huanglian jiedu decoction which absorbed in serum have been speculated.As a typical prescription, huanglian jiedu decoction have many pharmacology activity, including anti-cancer, anti-inflammation, et al. recently, many reports about huanglian jiedu decoction's effect on cardiovascular diseases has emerged. This assay explains the effects of huanglian jiedu decoction on the high-fat-diet induced hyperlipidemia rats for the first time, and we have also studied the regulation function of huanglian jiedu decoction on some relative gene in lipid metabolism. We have found that huanglian jiedu decoction could not only decrease the totel cholesterol, low density lipoprotein cholesterol and apolipoprotein B, but could also increase the high density lipoprotein cholesterol and total triglyceride level in the serum of hyperlipidemia rats. Compared with the Lipitor, a kind of drug which used clinically as a regulation lipid, huanglian jiedu decoction can significantly reduce the arteriosclerosis index, coronary heart disease index and ApoB/ApoA1(relative with miocardial infarction). Through the channel of raising the activity of superoxide dismutase and Glutathione peroxidase, reduced level of malondialdehyde, huanglian jiedu decoction promoted the ability of elimination free radical induced by hyperlipidemia, The activity of lipoprotein lipase and hepatic lipase have been increased by huanglian jiedu decoction to reduce the incident of as arteriosclerosis. Observed the pathological section of the liver, we noticed that huanglian jiedu decoction could meliorate the steatosis induced by high-fat-food.In order to explore the regulation function of huanglian jiedu decoction on the relative genes in lipid metabolism, we detected mRNA level of low density lipoprotein receptor (LDLR) and Peroxisome proliferator activated receptorγ(PPARγ) through the semi-quantitation polymerase chain reaction. The result of this experiment revealed that huanglian jiedu decoction could up-regulated the mRNA level of LDLR and PPARγ, and in this way the expression of these genes were changed. In order to explore the source of the active components in huanglian jiedu decoction, the effects of serum containing huanglian jiedu decoction and different combinations of drugs, berberine and baicalin on these gene was also be detected. The results showed that the serum concerning huanglian jiedu decoction or Scutellaria baicalensis Georgi plus Gardenia jasminoides Ellis could up-regulate the expression of LDLR and LXRα, suppress the down-regulation on PPARγ. These two sorts of serum could also inhibit the differentiation of 3T3-L1 cell. Meanwhile, in this experiment, we also find that berberine could up-regulate the LDLR and LXRαin HepG2 cell. However, because of the low concentration in huanglian jiedu decoction, the effect of berberine was limited. Baicaline have a high concentration in huanglian jiedu decoction, as a main compound in Scutellaria baicalensis Georgi, it has on effects on these gene. So we surmise, other compounds or the metabolic product of baicalin could regulate the lipid in serum, or baicaline effect the hyperlipidemia in other ways.At last, we used computer software as a screen platform, chose liver X receptor(LXR) as a target which drug destined to, and have docked 107 known compounds in the four drugs which composed huanglian jiedu decoction, in both of LXRs subtypes. Among these compounds, we found 6 compounds selective agonists to LXRβ, all these come from Scutellaria baicalensis Georgi and Gardenia jasminoides Ellis. Because of the stereospecific blockade, baicalin could not bind the LXRαwell, but showed a high bind ability to LXRβ. Through this way, baicalin can regulate lipid metabolism.