Mechanism Of Salvia Miltiorrhiza And Siwu Decoction On Anti-obesity By Regulating MAPK Signal Pathway

Objectives:By observing the effects of Salvia miltiorrhiza and Siwu Decoction on the body weight,physiological biochemistry,inflammatory response and the expression level of MAPK（Mitogen Activated Protein Kinase.MAPK）signaling pathway-related proteins in rats fed with high-fat diets,to explore the similarities in the efficacy of Salvia miltiorrhiza and Siwu Decoction.On the one hand,it can enrich the connotation of"one flavor of Salvia miltiorrhiza has the same function as Siwu Decoction"and promote the inheritance and development of the basic theory of Chinese medicine,on the other hand,it also expands the application scope of Salvia miltiorrhiza and Siwu Decoction and provides new directions and ideas for clinical development of weight loss drugs.Method:Ninety 7-week-old SPF-grade male SD rats with a body mass of（180±20）g were randomly divided into two groups:10 rats in the normal group and 80 rats in the model group,which were fed with normal diet and high-fat diet（D12451,45%fat content）respectively to construct the obese rat model.The modeling was completed when the mean body weight of the model group was 20%higher than that of the normal group,and then the model group was randomly divided into model control group（high-fat-diet group,HFD group）,Orlistat group(32.4 mg·kg^-1,Orlistat group),Salvia miltiorrhiza-low group(0.675 g·kg^-1,Salvia miltiorrhiza-Low,Sal-L group),Salvia miltiorrhiza-medium group(1.35 g·kg^-1,Salvia miltiorrhiza-Media,Sal-M group),Salvia miltiorrhiza-High group(2.7 g·kg^-1,Salvia miltiorrhiza-High,Sal-H group),Siwu Decoction-Low group(1.62 g·kg^-1,Siwu Decoction-Low,SWD-L group),Siwu Decoction-Medium group(3.24 g·kg^-1,Siwu Decoction-Media,SWD-M group),and Siwu Decoction-High group(6.48 g·kg^-1,Siwu Decoction-High,SWD-H group),the normal group is the normal control group（Control group）,and there are 9 groups,10 animals in each group.During the treatment,the normal control group and the model control group were gavaged with the same volume of saline according to their body weight every day,and each treatment group was gavaged with the corresponding drug according to the group once a day for 8 weeks,during which the changes in body weight were recorded weekly.After the last administration of the drug,the rats were fasted without water,and blood was collected from the abdominal aorta after12 hours of anesthesia,and the serum was separated to detect the contents of triglycerides（TG）,total cholesterol（TC）,high-density lipoprotein cholesterol（HDL-C）,low-density lipoprotein cholesterol（LDL-C）,aspartate Transaminase（AST）,alanine transaminase（ALT）,total bile acids（TBA）,glucose（GLU）,and free fatty acids（FFA）in the serum.Liver tissues and adipose tissues of rats were isolated,weighed,and the percentage of their body weight was calculated,and the pathomorphological changes of liver tissues and adipose tissues of rats were observed by HE staining.The levels of interleukin-6（IL-6）,interleukin-10（IL-10）and tumor necrosis factor（TNF-α）in liver tissues of rats were measured by ELISA,and the expression levels of MAPK signaling pathway-related proteins in liver tissues were measured by Western Blot.Results:1.Effects on body weight changes in rats:compared with the model control group,the body weight increase of each treatment group was smaller.After 8 weeks of treatment,the Orlistat group,Sal-L group,Sal-M group,Sal-H group,SWD-L group,SWD-M group and SWD-H group could significantly reduce the body weight of obese model rats（P<0.05,P<0.01）.2.Effects on liver-to-body ratio and lipid-to-body ratio of rats:compared with the model control group,Orlistat group,Sal-L group,Sal-M group,Sal-H group,SWD-L group,SWD-M group and SWD-H group could significantly reduce the lipid-to-body ratio of obese model rats（P<0.05,P<0.01）,and Orlistat group,Sal-M group and SWD-L group also significantly reduced the liver-to-body ratio（P<0.05,P<0.01）.3.Effects on blood lipid levels in rats:compared with the model control group,the levels of TG,TC and LDL-C in the Orlistat group decreased（P<0.05,P<0.01）;the Sal-M group was able to reduce the levels of TC and LDL-C in serum（P<0.05,P<0.01）,and the expression levels of TG did not have significant differences compared with the model control group（P>0.05）;the Sal-L group and Sal-H group was able to reduce the levels of TG in serum（P<0.05,P<0.01）,the expression levels of TC and LDL-C did not have significant differences（P>0.05）;the SWD-L group was able to reduce the levels of TC and TG in serum（P<0.05,P<0.01）,and the levels of LDL-C were not significantly different（P>0.05）;the SWD-M group was able to reduce the levels of TG and LDL-C in serum（P<0.05,P<0.01）,and the levels of TC were not significantly different（P>0.05）;Orlistat group,Sal-L group,Sal-M group,Sal-H group,SWD-L group,SWD-M group and SWD-H group could increase the level of HDL-C（P<0.05,P<0.01）.4.Effects on serum AST,ALT and TBA levels in rats:compared with the model control group,Orlistat group and SWD-M group were able to reduce the levels of AST,ALT and TBA in serum（P<0.05,P<0.01）;Sal-L group was able to significantly reduce the expression levels of AST and TBA in serum（P<0.05,P<0.01）,and The expression levels of ALT were reduced but there was no significant difference（P>0.05）;the Sal-M group could significantly reduce the expression levels of ALT and TBA in serum（P<0.05,P<0.01）,and the expression levels of AST were reduced but there was no significant difference（P>0.05）;the Sal-H group and SWD-L group could significantly reduce the expression levels of TBA in serum（P<0.05,P<0.01）,and the expression levels of AST and ALT were reduced but there was no significant difference（P>0.05）.5.Effects on GLU and FFA in rat serum:compared with the model control group,Orlistat group,Sal-L group,Sal-M group,Sal-H group,SWD-L group,SWD-M group and SWD-H group were able to reduce the levels of GLU in serum（P<0.05,P<0.01）,Orlistat group,Sal-L group,Sal M group,Sal-H group and SWD-L group were all able to reduce the levels of serum and FFA（P<0.05,P<0.01）.6.Effects on the pathological morphology of liver and adipose tissue of rats:in HE staining of liver tissue,the normal control group had normal hepatocyte morphology,no steatosis and inflammatory reaction was seen;the model control group saw obvious hepatocyte vacuolation with larger vacuole area and high number;hepatocytes in each treatment group were significantly improved compared with model control group vacuolation,with smaller vacuole area and reduced number.In the HE staining of adipose tissue,the overall outline of adipocytes in the normal control group was smaller and more numerous;the outline and volume of adipocytes in the model control group became larger and loosely arranged.Compared with the model control group,the outline and volume of adipocytes in each treatment group became smaller and more closely arranged.7.Effects on IL-6,TNF-αand IL-10 in rat liver tissues:compared with the model control group,Orlistat group,Sal-M group,SWD-L group,SWD-M group and SWD-H group could significantly reduce the levels of IL-6 and TNF-α（P<0.05,P<0.01）and elevate the level of IL-10 in liver tissues（P<0.05,P<0.01）.8,Effects on the expression levels of MAPK signaling pathway-related proteins in rat liver tissues:compared with the model control group,the Sal-H,SWD-L,SWD-M and SWD-H groups were able to down-regulate the expression levels of p38MAPK,p-p38MAPK,ERK1/2and PPARγproteins（P<0.05,P<0.01）,and the Sal-L group could elevated the protein expression levels of p-ERK1/2,JNK（P<0.05,P<0.01）,the Sal-H group could elevated the protein expression levels of p-ERK1/2,p-JNK（P<0.05,P<0.01）,and SWD-L group elevated the protein expression levels of JNK,p-JNK（P<0.05,P<0.01）.Conclusion:1.According to the analysis of the results of this experiment,different doses of Salvia miltiorrhiza and Siwu Decoction have effects on anti-obesity caused by high-fat diet feeding in rats.Among them,the effect of middle dose of Salvia miltiorrhiza was better,followed by high dose;the effect of low dose of Siwu Decoction was better,followed by middle dose.2.The mechanism of"anti-obesity"of Salvia miltiorrhiza and Siwu Decoction may be related to the reduction of blood lipid level,regulation of MAPK signaling pathway and inhibition of PPARγand its downstream protein expression.