Effects Of Jianpifang On Insulin Resistance In Muscle And Adipose Tissue Of T2DM Rats Based On The Spleen Being In Charge Of The Muscles And PGC-1α-FNDC5/Irisin Signaling Pathway

Objective:To investigate the effects of Jianpifang on glucose and lipid metabolism,serum irisin level,skeletal muscle and adipose tissue morphology,PGC-1α-FNDC5/Irisin signal-related protein and mRNA expression in T2DM rats,On the intervention of muscle and fat IR in T2DM rats,and to elucidate its mechanism.It aims to provide effective prescription for the treatment of T2DM and provide experimental support for the treatment of T2DM muscle and fat IR from spleen.Methods:1.Theoretical study:This study combs the concept,characteristics and function of spleen,analyzes the mechanism of spleen function,the mechanism of spleen deficiency causes diabetes,and the theoretical basis of spleen deficiency causes diabetes.On the basis of this,the connotation of "Spleen being In charge of the muscles" was studied from the perspective of integrated traditional Chinese and western medicine,and the energy metabolism unit of "spleen-skeletal muscle-fat" was established,and the state of phlegm and blood stasis due to deficiency of spleen is the importance Mechanism of the IR in muscle and fat,and discuss the relationship between spleen-skeletal muscle-fat unit and PGC-1α-FNDC5/Irisin signaling pathway.2,experimental study(1)54 Wistar rats,taking 10 as normal control group,44 high-sugar high-fat diet 6 weeks after intraperitoneal injection of STZ established T2DM model,modeled rats after random model group(13 rats),metformin group(14 rats),traditional Chinese medicine group(14 rats),normal group and model group were given pure water gavage,metformin group given metformin suspension,traditional Chinese medicine group given Jianpifang.After 6 weeks,fasting blood glucose,blood lipid,glycosylated hemoglobin,serum irisin and serum insulin were calculated.HOMA-IR was calculated and insulin resistance test was performed.The gastrocnemius,white fat and brown adipocyte morphology were observed by HE staining.The expressions of PGC-la,FNDC5,GLUT4 in skeletal muscle,PGC-1α,FNDC5 and UCP1 in adipose tissue were observed by Western Blot.The expression of PGC-1α,FNDC5,GLUT4 mRNA in skeletal muscle,PGC-1α,FNDC5 and UCP1 mRNA in adipose tissue was detected by RT-PCR.Result:The effect of Jianpifang on glucose and lipid metabolism,islet function and insulin sensitivity in T2DM rats: Compared with the normal group,the blood glucose level,fasting insulin level and HOMA-IR in the model group were significantly higher than those in the normal group(P<0.01),and the levels of TG,TC,LDL and FFA were increased and HDL(P<0.01).Compared with the model group,the levels of serum irisin were significantly decreased(P<0.01).Compared with the model group,the metformin group and the traditional Chinese medicine group could decrease the blood glucose(P<0.05)(P<0.01).In the aspect of improving the IR,the spleen squared with metformin was equivalent(P>0.05).In the regulation of dyslipidemia,compared with the model group,the levels of TG,TC,LDL and FFA were decreased in the metformin group and the traditional Chinese medicine group,among them the traditional Chinese medicine group to reduce FFA,increased the role of HDL is more obvious(P<0.05).The level of serum irisin in the treatment group was higher than that in the model group(P<0.05).2、The effect of Jianpifang on PGC-1α-FNDC5/Irisin signaling pathway in muscle and adipose tissue of T2DM rats:Compared with the normal group,the cell morphology of gastrocnemius muscle cells in the model group was confused and the cell gap increased,the expression of PGC-la,FNDC5,GLUT4 protein and mRNA were significantly decreased(P<0.01).White fat cells ranging in size,disorder,cell boundaries are unclear,and there is a fusion phenomenon,There was no significant difference in PGC-la expression between the two groups(P>0.05),The mRNA and expression of FNDC5 protein were up-regulated(P<0.01).Brown fat cells in the cytoplasm of large fat vacuoles more;gastrocnemius cells,brown fat cells,PGC-1α,FNDC5,UCP1 protein and mRNA expression were significantly decreased.Compared with the model group,the gastrocnemius and adipocytes in the drug intervention group were more regular and arranged in a more regular and clear cell boundaries.The PGG-1α,FNDC5,GLUT4 protein and mRNA expression were increased in the metformin group and the traditional Chinese medicine group(P<0.01(P<0.01).The expression of FNDC5 and mRNA in white fat was decreased(P<0.01).The expression of PGC-1α,FNDC5 and UCP1 protein and mRNA in brown fat were decreased(P<0.01).Conclusion:1.theoretical research:This study is based on the "Spleen being In charge of the muscles ", proposed from the spleen on the treatment of Skeletal muscle and adipose tissue insulin resistance,raised that the phlegm and blood stasis due to deficiency of spleen is important pathogenesis of T2DM muscle and adipose tissue IR,established"invigorating the spleen and replenishing qi,resolving phlegm and promoting blood" the principle of treatment,composing the jianpifang and for the clinical prevention and treatment of IR provides an effective way and drugs;And in-depth study of the mechanism of spleen deficiency to create a "spleen-skeletal muscle-fat" unit,combined PGC-1α-FNDC5/Irisin signaling pathway,suggesting that there is a close correlation between the two.2.Experimental study:Jianpifang can reduce the blood glucose level of T2DM rats,regulate lipid metabolism disorders,enhance insulin sensitivity,improve serum irisin levels,protect and partially repair T2DM rats skeletal muscle and adipose tissue structure and function,improve Muscle and adipose tissue IR,the mechanism may be related to the up-regulation of PGC-1α-FNDC5/Irisin signaling pathway-related protein expression.Through the above experiments to support the treatment of muscle and adipose tissue IR from the spleen is effective,to verify the Jianpifang in improving the 1R,the exact role of treatment of T2DM,and provide effective treatment of drugs.