| Objective This study aimed to investigate the role and mechanism of Guanxin Danshen Dropping Pill on improving IR from the perspective of GLUT-4,by exploring effects of the drug on biochemical metabolic markers such as Type II diabetes HOMA-IR,as well as GLUT-4 expression and cell membrane translocation in skeletal muscle and cardiac muscle cells.Method Type 2 diabetes IR rat models were established,that were administered with the drug.Afterwards,changes of skeletal muscle and cardiac muscle cell GLUT-4expression and cell membrane translocation in phosphoinositide 3-kinase(PI3K)signaling pathway were studied.In vitro study: palmitic acid(PA)-induced L6 cell models of IR rats were used,to which the druy were applied,the drug's effects on GLUT-4 mRNA expression and cell membrane translocation in skeletal muscle and cardiac muscle IR models were studied.L6 skeletal muscle cells were used as subjects for in vitro study.Impacts of Guanxin Danshen Dropping Pill on skeletal muscle cell viability were detected and appropriate concentration of the drug working on in vitro cell culture was determined by MTT assay.IR skeletal muscle cell models were established by inducing rat L6 cells with palmitic acid.After being applied Guanxin Danshen Dropping Pill in the cells,glucose concentration of supernatant was detected by GOP-POD process,impacts of the drug on glucose uptake in IR cells were studied;triglyceride concentration and FFA content in cells were both detected by enzymatic process,impacts of the drug on lipid deposition in IR cells were studied;and impacts of the drug on GLUT-4 protein expression in IR cells were studied by Western Blot blotting.Diabetic rats were used as subjects in animal experiment,rats were sacrificed after Guanxin Danshen Dropping Pill administration,venous blood was collected and centrifuged,then biochemical markers including serum insulin level,cholesterol concentration,glucose concentration,triglyceride concentration and glycated hemoglobin level were detected,impacts of the drug on IR associated markers in diabetic rats were studied;in addition,GLUT-4 locations in skeletalmuscle and cardiac muscle tissues were observed by immunohistochemical techniques,and optical densities of GLUT-4 indicating protein contents were determined,therefore the mechanism of Guanxin Danshen Dropping Pill on improving insulin resistance was investigated.Result Effects of Guanxin Danshen Dropping Pill on cell or animal models were studied via establishing insulin resistance models by culturing in vitro skeletal muscle cells.In cell experiment,effects of the drug on cell IR models were studied as following.After culturing in vitro skeletal muscle cells and inducing L6 muscle cells IR with palmitic acid,the drug were applied.Then a significant increase in glucose uptake of L6 muscle cells was observed,extent of IR in PA-induced L6 muscle cells was improved;contents of TG and NEFA were reduced in L6 muscle cells,a reduction in lipid deposition in PA-induced L6 muscle cells was also noticed.Similar conclusions were reached by establishing diabetic animal models;besides,blood glucose can be reduced and blood lipid function can be improved by the drug.Immunohistochemical methods further confirmed that,GLUT-4 expression in skeletal muscle and cardiac muscle tissues was increased with Guanxin Danshen Dropping Pills,indicating the drug's efficacy in improving IR through GLUT-4 pathway.GLUT-4 is widely found in insulin sensitive skeletal muscles,glucose transport mediated by it plays a vital role in regulating glucose homeostasis.An increase in GLUT-4 expression was noticed after the drug application,thereby glucose uptake was facilitated and insulin resistance was improved.Conclusion By adjusting some critical factors influencing IR,Guanxin Danshen Dropping Pill facilitates GLUT-4 translocation,thereby increasing GLUT-4 expression.We preliminarily verifies that the drug play a role in IR,thus increase insulin sensitivity,ameliorate blood glucose and reduce blood lipid. |
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