| [Objective] Diabetic nephropathy (DN) that has been an important cause of death in diabetics is one of the common microvascular complications of diabetes. The incidence of DN has increased year by year. The principal of pathological changes of DN included glomerulus hypertrophy, basement membrane thickening and glomerulus extracellular matrix accumulating in the early stage; glomerular and renal interstitial fibrosis in late. The pathogenesis of diabetic nephropathy is related to many factors. Many cytokines affecting the hemodynamic change, extracellular matrix, cell proliferation, hypertrophy, and many other aspects were considered to be an important aspect of DN.Tangshenkang traditional Chinese medicine that had required good effect in the treatment of diabetic nephropathy has been used in clinical for many years. It not only can reduce cholesterol, improve renal function, reduce microalbuminuria (MALB), also can reduce the nitric oxide levels in the rat model of diabetic nephropathy. Researching the effect of cytokines expression and TGF-β1/Smad2/3signaling pathways in renal mesangial cells (RMCs) under high glucose which is caused by Tangshenkang serum containing traditional Chinese medicine and exploring the mechanism of traditional Chinese medicine in curing diabetic nephropathy provide a certain basis and foundation for clinical treatment.[Methods]12SD rats were randomly divided into four groups:low dose group, middle dose group, high dose group, control group. After7-day gavage via portal vein blood, the rats were sacrificed and serum was collected to culture the RMCs with different concentration of serum containing traditional Chinese medicine and blank serum. Cell proliferation was determined by methly thiazolyl tetrazolium (MTT) assay to determine the optimal serum concentrations of medicine; Culture the RMCs With the best blood drug concentration of serum. The expression of cytokines was testing by Protein chip; TGF-β1protein expression was determined by ELISA; Smad2/3protein expression and phosphorylation was determined by Western blotting and Immunofluorescence.[Results]1.There is no significant difference in cell survival rate between the control group and different concentration traditional Chinese medicine serogroup (P>0.05); the increment rate of high sugar1.40times as the control group (P<0.05); compared with the high sugar group, the cell proliferation of the traditional Chinese medicine serum groups were significantly lower (P<0.05); the cell proliferation of the low dose of Chinese traditional medicine serum group was significantly inhibited compared with the middle dose of Chinese traditional medicine group and the high dose of Chinese traditional medicine group(P<0.05); the cell proliferation has no obvious difference between the middle dose Chinese traditional medicine group and high dose traditional Chinese medicine group (P>0.05).2. Protein chip shows that the expression of TGF-β1, IL-1, IL-6, TNF-a, VEGF was significantly increased in the high glucose group compared with the control group; The expression of these factors was significantly reduced in the Chinese medicine group compared with the high glucose group.3. The expression of TGF-beta1in the Chinese medicine group was decreased compared with the high sugar group by the ELISA method (P<0.05). The expression of p-Smad2/3of the high sugar group was significantly increased (p<0.05) compared with control group, while it was significantly decreased of the Chinese medicine serum group (p<0.05).(Conclusion]:The serum containing Jian-Pi-Yi-Yin-Huo-Xue Chinese Herbal compound can inhibit the proliferation of RMCs under high glucose by regulating the expression of cytokines. The main mechanism may be associated with the inhibition of TGF-β1/Smad2/3pathway. |
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