An In Vitro Screen Of Traditional Chinese Medicine Attenuating Adipocyte Proliferation Via Regulating The Expression Of PAI-1 Gene

Objectives:With the development of the society, obesity is vitally important to threat human health,especially in our country。While the mechanism study of obesity etiological is still unclear,the putative reason course the chaotic of energy is the inexpedient collocation of diet,genetic mutations,deficient activities and the enviroment of the society and so on,A large quantity of study shows that the standard of PAI-1 in human body is relevant to obesity.This experiment will induced 3T3-L1 adipocytes into mature 3T3-L1 adipocytes. On the basis of PT-PCR and plasmids transfection,we attempt to Study the relationgship between traditional Chinese medicine and the expression of PAI- 1 gene.We tried to study the effect of Chinese medicine preparations for obesity mechanism from the level of molecular biology for clinical treatment of obesity.Methods: The experiment is according to the following steps.First of all,we choose six traditional Chinese medicine to make the following subsequent trials through the literature retrieval.We extracte liquids to concentrat to 1 g/ml, packing standby in 4 degrees.Then we induce 3T3-L1 adipocytes into mature 3T3-L1 adipocytes by the classic method of cocktails for the preparation of the follow-up test. We stimulate the mature 3T3-L1 adipocytes with 1 mg/ml of TCM in 24 h and extract RNA.Then we detect the influence of traditional Chinese medicine preparation respectively on PAI- 1 mRNA level with mature 3T3-L1 adipocytes though the real-time PCR technology. At last,we Build luciferase reporter gene carrier containing PAI- 1 promoter fragment though cloning PAI- 1 gene promoter fragment.Then we transfect the recombinant plasmid into mature 3T3-L1 adipocytes.We stimulate the cell model with 1 mg/ml of TCM 24 hours.At last,we select the TCM which have the influential expression of PAI- 1 promoter.Results: Compared with normal group, weight loss capsules, wind through the solution and alisma soup’s have the statistically significant differences on the expression of PAI-1mRNA with mature 3T3-L1 adipocytes by RT-PCR technology.The rest of the medicine has no obvious significance.Transfection after 48 hours mediated by liposome, the luciferase activity of pGL3- Basic- 1.1 have the statistically significant compared with pGL3-Basic.So we choose pGL3- Basic- 1.1 for subsequent tests.Compared with normal group, weight loss capsules, and wind through the solution have the statistically significant by detecting the luciferase activity of PAI-1 promoter.Conclusions:Weight loss capsules, windproof tong SAN and alisma soup have the function to induced PAI- 1 mRNA expression.The cells model which successful contain PAI- 1 promoter transient transfection could screen out the TCM that have the influence on the luciferase activity of PAI-1 promoter.At last,we select the weight loss capsules, and windproof tong SAN Inhibitted the activity of PAI- 1 gene promoter.