| Background:"Formula-factors refers to the elements of herbal medicine,dosages,dosage forms,and functions that make up the formulas.These elements have clear purposes and directions.The thinking mode of the clinical prescriptions that associates the "formula factors" with the"syndrome-factors" is the theory of "correspondence between syndrome and formula factors".Dachaihu decoction can reconciliate both Shaoyang and Yangming in "Treatise on Febrile Diseases" and "Synopsis of Golden Chamber",and treat not only the unsmooth qi activity of liver and gallbladder,liver and spleen discord caused by liver wood subjugates earth,but also phlegm obstruction in the stomach or the syndrome of Yangming Fu-organ disease.According to its basic pathogenesis,there are three basic syndrome-factors called "unsmooth qi activity of liver and gallbladder","spleen deficiency with dampness stagnated" and "failure of Yangming dreggy qi to desend" in Dachaihu decoction.The formula-factors of "Shugan Lidan"are consisted of bupleurum,scutellaria baicalensis and radix paeoniae alba,corresponding to the syndrome-factors of "unsmooth qi activity of liver and gallbladder".Nonalcoholic fatty liver disease(NAFLD)is currently the largest chronic liver disease in China.The lesions are mainly based on the hepatic lobules,and the main pathological changes are liver cell steatosis caused by excessive liver lipid accumulation.There are numerous pathogenic factors and the pathogenesis is complex.Lesion is often accompanied by insulin resistance(IR)and oxidative stress,the specific manifestations are abnormal liver function,as well as disorders of lipid metabolism and glucose metabolism.The disease spectrum of NAFLD includes nonalcoholic simple fatty liver(NAFL),nonalcoholic steatohepatitis(NASH)and its associated cirrhosis and associated hepatocellular carcinoma.Liver controls dispersion,and promotes the qi activity and gasification of the whole body.If it fails to disperse,it will cause stagnation.Gallbladde controls descent.If fails to descend,it will cause symptoms of pain,swelling,and congestion.Qi activity not only can make changes in the movement of substances inside and outside the body in the metabolic process,but also maintain coordination during this process.Objective:This research includes three parts:literature review,theoretical discussion and experimental research.To Simulate the occurrence and evolution of human NAFLD by Establishing the Rat Model of NAFLD with a high-fat and high-sugar Diet,and intervened by the formula-factors of "Shugan Lidan" and Dachaihu decoction.Observing the effects of liver function,lipid and glucose metabolism by the formula-factors of "Shugan Lidan",to discuss it’s main action targets,so as to further understand the prescription structure of Dachaihu decoction and provid experimental evidence for its clinical application.Methods:The 40 SD rats were randomly divided into 4 groups:normal group,model group,Dachaihu decoction group,and Shugan Lidan group.There are 10 rats in each group.Rats in the normal group were feeded by fundamental forage,and the remaining three groups were given high-fat and high-sugar Diets.During the experiment,each group was given daily feed water,and the daily feed amount of each group of rats was recorded,as well as the rats’ daily dietary intake was calculated.The body weight of rats was recorded regularly and the growth condition of rats was observed last 16 weeks.Modeling period from the 13th week to the 16th week,the rats of Shugan Lidan group and Dachaihu decoction group were fed daily by the Shugan Lidan formula-factors(bupleurum,scutellaria baicalensis and radix paeoniae alba)3ml and Dachaihu decoction 3ml.Rats of the normal and model groups were given equal amounts of distilled water at the same time,once daily for 28 days.At the end of 16th week,collected the tail blood at 12h after the last intragastric administration.Used blood glucose meter to measure fasting blood glucose(FBG)levels.After weighing,all rats were given intraperitoneal injection of 10%chloral hydrate 3ml/kg,drew abdominal aorta blood and dissected the liver rapidly.Observed the liver appearance of each group’s rats visually and weighed the liver wet weight to calculate the liver index.Made frozen sections and paraffin sections.Used the staining method of HE,oil red O and Masson to judge the staging of NAFLD disease,by observing histopathological changes of hepatocytes under light microscope.Measured the levels of each group of rats’ serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),total cholesterol(TC),triglyceride(TG)and high-density lipoprotein cholesterol(HDL-C),lowdensity lipoprotein cholesterol(LDL-C),and calculated the ratio of TG/HDL-C.Results:1.General situation:The rats of model group was thinner than the normal group,with less food and water intake,restlessness,and the hair is yellow,unkempt,dull,with severe exfoliation of tail scales.4 weeks after intragastric administration,compared with model group,there was no significant change in dietary and drinking water in the Shugan Lidan group and Dachaihu decoction group,and the rest of the conditions were improved.2.Liver condition:Compared with the normal group,rats in model group had the following features:liver volume increased significantly and the color was yellowish brown,as well as the capsule was tight and lackluster,It had full round blunt edge and it’s texture was more rigid,It’s elasticity was poor snd the cut surface was greasy.Compared with the model group,rats in Shugan Lidan formula-factors group and Dachaihu decoction group had the following features:liver volume reduced and the color was light yellow,as well as the tight capsule was reduced and turned smooth,it’s texture was rigid,It’s elasticity was slightly worse and the cut surface was greasy.3.Liver index:The rats’liver index of the model group were significantly higher than those in the normal group,the difference was statistically significant(P<0.01).After 4 weeks of intragastric administration,the liver index of the Shugan Lidan and Dachaihu decoction groups was lower than that in the model group.The difference was statistically significant(P<0.05 and P<0.01).The liver index of the Shugan Lidan group was higher in the Dachaihu decoction group,and the difference was statistically significant(P<0.05).4.Comparison of liver pathological changes:(1)Liver tissue structure and cell morphology:HE staining results show that compared with the normal group,the model group’s structure of the hepatic lobule was disordered,and the hepatocytes were swollen and deformed,with a small number of hepatocytes balloon changed.And the large-bubble and small-bubble mixed lipid droplets were accumulated in the cytoplasm,with the cell nucleus’ position were moved.Compared with the model group,the liver lobular’s structure was clearer in Shugan Lidan group and Dachaihu decoction group,with hepatocyte swelling and deformation significantly reduced,significant decrease in ntracellular lipid droplets.(2)Observation of liver tissue steatosis:Oil red O staining result display that compared with the normal group,liver cells of the model group in the visual field are flooded with a large number of orange-red and large vesicle-type lipid droplets,fatty cells were diffuse and distributed,with the nucleus dissolved and blurred.The liver cell boundaries between the Shugan Lidan group and Dachaihu decoction group were more clear than that of the model group,with the numbers of lipid droplets in the cell were reduced,and the degree of accumulation of orange-red lipid droplets in the visual field is significantly reduced,large flakes of lipid droplets occasionally.(3)Observation of inflammatory cell infiltration and fibrosis in liver tissue:Masson staining showed no obvious inflammatory cell infiltration in both four groups,as well as fibers in the prefecture area had not been expanded and increased,no fiber and liver cell degeneration and necrosis appeared.(4)NAFLD disease stage:NAFLD did not occur in normal group.And the model group suspicious for non-alcoholic steatohepatitis(NASH),with Shugan Lidan and Dachaihu decoction were simple fatty liver.5.Liver function:The serum ALT and AST levels in the model group were significantly higher than those in the normal group(P<0.01).The serum levels of ALT and AST in the Shugan Lidan group were lower than those in the model group,the difference was statistically significant(P<0.05 and P<0.01).Dachaihu decoction group rats’ serum ALT,AST significantly reduced than the model group,(P<0.01).Compared with Dachaihu decoction group,the levels of ALT and AST in serum of Shugan Lidan group were higher,and the difference was not statistically significant(P>0.05).6.Blood lipids:Compared with the normal group,serum TC,TG,LDL-C and TG/HDLC ratios in the model group were significantly higher,and HDL-C was significantly lower(P<0.01).Compared with the model group,TC of the Shugan Lidan group decreased(P<0.05),TG,LDL-C and TG/HDL-C decreased significantly,HDL-C increased significantly,the difference was statistically significant(P<0.01).Compared with the model group,the serum TC,TG,LDL-C and TG/HDL-C ratios in Dachaihu decoction group were significantly lower,and HDL-C was significantly higher(P<0.01).Serum LDL-C in the Shugan Lidan group was slightly higher than Dachaihu decoction group(P<0.05),differences in the remaining blood lipids were not statistically significant(P>0.05).7.Blood glucose:The rats of the model group were significantly higher than those in the normal group(P<0.01).The rats in Shugan Lidan group and Dachaihu decoction group were significantly lower than those in the model group,and the difference was statistically significant(P<0.01).Compared with the Dachaihu decoction group,the blood sugar level of the Shugan Lidan group was higher,and the difference was not statistically significant(P>0.01).Conclusion:1.The Rat Model of NAFLD successfully induced by high-fat and high-sugar diet in this study,was similar to the characteristics of liver lesions and changes in glucose and lipid metabolismin of modern human NAFLD caused by the daily diet.2."Shugan Lidan" formula-factors(bupleurum,scutellaria baicalensis and radix paeoniae alba)in Dachaihu decoction can improve liver function and regulate glucose with lipid metabolism in NAFLD rats.It prompt that the main action target of the "Shugan Lidan"formula-factors is on the liver,and it is closely related to regulating liver function and glucose lipid metabolism,during the treatment of NAFLD by Dachaihu decoction.In order to further understand the prescription structure of Dachaihu decoction and provid experimental evidence for its clinical application. |
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