| The central role played by Chinese herbal pieces in the clinic of Chinese medicine is increasingly apparent,but the theory of traditional Chinese medicine(TCM)processing which guides the reasonable application of Chinese herbal pieces still lacks modern scientific arguments,and it has become a bottleneck problem that hinders the modernization of TCM which must be solved urgently.Schisandra chinensis processed with vinegar has already achieved significant clinical curative efficacy in the treatment of liver disease,but the research has not seen on the mechanism of its hepatoprotective effect.The key mechanisms of hepatoprotective effect of medicines were related to metabolic enzymes and signaling pathways.According to the above issues and the existing research foundations,we introduced cooktail probe drug laws,Real-time PCR and western blot and so on frontier technologies to the processing theory of Chinese herbal medicine field innovatively.Taking Nrf2/ARE pathway as the starting point,to evaluate the correlation of metabolic enzyme,metabolic component and the body.We will study and clarify the scientific connotation of the hypothesis that Schisandra chinensis processed with vinegar expressed a hepatoprotective effect by regulating Nrf2/ARE pathway in organism from the the overall and cellular levels.The main contents are all of follows:1.Schisandrin chinensis was chose in our experiment and its before and after being processed with vinegar of quality assessment.Using Schisandrin chinensis as raw material which quality to meet the requirements of the 2010 edition of a Chinese Pharmacopoeia to process raw and vinegars products.Simultaneous determination of 8 lignans in Schisandra chinensis before and after being processed with vinegar by quantitative analysis of multi-components by single marker related quality assessment.The results showed schisandrin chinensis before and after being processed with vinegar lignans components have different degrees of change.The contents of Schisandrin,Gomisin J.Schisandrol B,Gomisin G,Deoxyschisandrin.Schisantherrin A,Schisandrin B.Schisandrin C of proportion in raw products were 22.11:2.06:7.60:1.00:0.50:4.95:14.60:2.45 while vinegars products have become 23.61:1.74:5.42:1.00:1.14:6.57:11.89:1.29.Tips the ratio and amount of lignin monomer components had corresponding changes after processed by vinegar.Another study showed that before and after the vinegar Schisandra chinensis lignans total contents were basically unchanged.Suggesting that there may lignans unknown monomer component increases or generates a new lignans monomer.This change results not only provide for the specific quality evaluation methods schisandrin chinensis before and after being processed with vinegar a reliable basis,but also probably after Schisandra hepatoprotective effect of vinegar to enhance one of the important reasons,to provide an effective system for the traditional vinegar Theory scientific basis.2.The effect of the alcohol extract of the raw and vinger processed schisandrin chinensis to acute alcohol-induced liver damage.Experimental research of group early on acute alcoholic liver injury did effects evaluation.Using the eliasa and ultraviolet spectrophotometer to detect the value of serum ALT,AST,SOD,ADH,TBIL,CHO,TG,HDL,LDL and SOD,MDA,ADH on liver to evaluate the effect of liver protection of the alcohol extract of the raw and vinger processed schisandrin chinensis.The results showed that both raw and vinger processed schisandrin chinensis have the effect of liver protection,which shown the significant difference compared with the vinger and show the dose-dependent effect(P<0.05).On this basis,the expression mRNA levels and protein levels of CYP2E1,Nrf2,HO-1,GCLM and NQOl in liver of ras respectively were detemined by Real-time PCR,Western blot method and Immune histochemical method.3.The effect of the alcohol extract of the raw and vinger processed schisandrin chinensis to chronic alcohol-induced liver damage.The chronic hepatic injury model was established by intragastrically administered with gradient alcohol for 8 周 eeks.The pathological section have shown that the model of acute alcoholic-induced liver damage was established successfully.The liver function,blood lipid levels and oxidative-antioxidant capacity changes in serum and liver tissue of liver of rats were measured by eliasa and ultraviolet spectrophotometer and HE staining of liver pathology observed.Histology manlfestion paralleled to biochemical findings.In the control group,constitution of hepatic lobules was normal,central veins was big and had thin paries;.Hepatic cells were polygon which were arranged in hepatic cord,which were distributed as radial around central veins.Infiltrating in portal regions in control rats.Abnormal hepatic lobules featuring by severe fatty degeneration,focal and regional necrosis of hepatic cells,inflammatory cells infiltrate in quantity,mesenchyma fibrosis could be seen in model rats.The groups of the raw and vinger processed schisandrin chinensis significantly reduced the levels of AST,ALT,AKP,TG,TC,LDL-C.TNF-α,IL-β,IL-6,ROS,MDA in serum and liver tissue and the levels of GGT,LDH in serum which were induced by ethnol and significantly induced the levels of HDL-C,SOD,GSH,GSH-Px,CAT,ADH in serum and liver tissue which were reduced by ethnol.All of treament groups showed alleviation in rat liver injury to varying degrees.Conclusion:The results indicated that the raw and vinger processed schisandrin chinensis both had a protective effect on chronic alcohol liver injury in rat and the mechanism of it may relate to the anti-oxidant effects which shown the significant difference compared with the vinger and show the dose-dependent effect.4.The mechanism of the alcohol extract of the raw and vinger processed schisandrin chinensis to acute and chronic alcohol-induced liver damage.In this study,the main idea is liver specific role by studying the Nrf2/ARE signaling pathway and alcohol metabolism of CYP2E1 enzyme analysis of biological information to evaluate the mechanism of schisandrin chinensis after being processed with vinegar.Rat acute and chronic alcoholic liver injury model was established,this experiment intended to study in liver tissue level.The expression levels of CYP2E1,Nrf2,HO-1,GCLM and NQO1 mRNA in liver of ras were detemined by Real-time PCR.The expression levels of Nrf2 nucleoprotein and CYP2E1,Nrf2,HO-1,GCLM and NQO1 cytoplasmic protein were detemined by Western blot method and Immune histochemical method.Distribution detect protein expression of the corresponding index in the liver nuclei or cytoplasm of liver cells using immunohistochemical staining experiments for the corresponding authentication.The results showed that groups of the raw and vinger processed schisandrin chinensis significantly reduced the levels of CYP2E1 mRNA and cytoplasmic protein which were induced by ethnol and significantly induced the levels of CYP2E1,Nrf2,HO-1,GCLM and NQO1 mRNA and cytoplasmic protein which were reduced by ethnol.In additional,all kinds of groups had no significant effect on the expression level of Nrf2 cytoplasmic protein by Western blot and immunohistochemical staining experiments.The results indicated that the raw and vinger processed schisandrin chinensis both had a protective effect on chronic alcohol liver injury in rat and the mechanism of it may relate to the Nrf2/ARE signaling pathway and alcohol metabolism of CYP2E1 enzyme effects which shown the significant difference compared with the vinger and show the dose-dependent effect.Conclusion:In summary,this subject selected schisandrin chinensis after being processed with vinegar as studied object.The proportion of lignins in schisandrin chinensis after being processed with vinegar occurs changes,the acute alcoholic liver injury and chronic alcoholic liver injury pharmacological effect and mechanism were compared.Under the guidance of Chiniese medicine theories,we combined multiple modern methods in our research,such as western blot,immunohistochemical staining experiments,Real-time PCR.Both the research ideas and technologies were innovative.The results could further explain the mechanism of liver channel tropism of vinegarprocessing schisandrin chinensis.It also provides new ideas for the modernization research of TCM. |
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