ABSTRACT
Objective:To study the effect of the prescription Tangshenning on the proliferation of the high glucoseinduced cells of near-end kidney tubules.Method:To prepare durg-contained serum from mice to enter into in vitor reaction system,the cells were randomly divided into 4 groups:the normal group,the model group,the,the irbesartan group,the tangshenning group and then detect the effects of all serum sections on the proliferation of high glucoseinduced epithelial cells of kidney tubules by the MTT colorimetric method,and the expression of RhoA,ROCK 1,Ecadherin and α-SMA in each group were detected by Western blotting.Result:The high glucose group of renal tubular epithelial cells form from the flat irregular polygon into long fusiform;After adding the drug-containing serum corresponding intervention into the cells into a flat in irregular polygon.The high glucose group of renal tubular epithelial cells show obvious proliferation condition,In the condition of 24 h,48 h,The high glucose group proliferation is better than the normal group (P＜0.01),in 60 h,The high glucose group proliferation is better than the normal group (P＜0.05);In 24 h,compared with the irbesartan,Tangshenning has better effect of inhibiting the cell proliferation(P＜0.05);In 48 h,Tangshenning has the better effect than irbesartan and Y27632 (P＜0.01);In 60 h,Tangshenning has the better effect than irbesartan and Y27632 (P＜0.05);Western blotting:Western blotting analysis showed that compared with the normal group,the expression of RhoA protein in high glucose group and Y27632 decreased (P＜0.01);The high glucose group and Tangshenning have significant difference (P＜0.01);Y27632 and Tangshenning have difference (P＜0.05).compared with the normal group,the expression of ROCK1 protein in high glucose group decreased (P＜0.01);The high glucose group,Tangshenning,the irbesartan and Y27632 have difference (P＜0.05).Compared with the normal group,the expression of a-SMA protein in high glucose group decreased (P＜0.01);The high glucose group,Tangshenning,the irbesartan and Y27632 have difference (P＜0.05).Compared with the normal group,the expression of E-Cadherin protein in high glucose group increased (P＜0.05).Y27632 and Tangshenning have difference (P＜0.05).The high glucose group,Tangshenning,the irbesartan and Y27632 have difference (P＜0.05).Conclusion:The prescription Tangshenning is able to inhibit the proliferation of high glucose-induced epithelial cells of kidney tubules and and can reverse renal tubularepithelial cell transdifferentiation via regulating RhoA/ROCK signaling pathway,and restrain renal interstitial fibrosis,thereby delaying the pathogenesis of diabetic kidney disease.
ABSTRACT
This paper was aimed to study the renal protective effect of Tang-Shen-Ning (TSN) on diabetic nephropathy (DN) KKAy mice by inhibiting the Notch/snail1 signal transduction pathway.A total of 30 KKAy mice,which were fed with mice-dedicated food for 10 weeks and with the blood glucose over 16.7 mmol· L-1,24-hour urinary albumin larger than 0.4 mg,were made into the DN model.The DN mice were randomly divided into the model group,irbesartan group and TSN group according to their blood glucose and weight.Intragastric administration of medication was given.A total of 10 female C57BL/6J mice were selected as the control group.The general condition,body weight and 24-hour urinary protein quantitation were detected.After 16-week intervention,mice were sacrificed.Levels of blood glucose,blood urea nitrogen (BUN) and serum creatinine (Scr) were detected.HE and Mallory staining were applied to renal tissues.In situ hybridization (ISH) and western blotting were used to detect the Notch/snail 1 pathway,α-SMA,E-Cadherin protein and mRNA expression in renal tissues.Statistical analysis was made by SPSS20.0 software.The results showed that compared with the model group,the rats' general conditions were improved;body weight and 24-hour urinary protein quantitation were significantly decreased (P＜0.01);contents of BUN and Scr were reduced (P＜0.01,P＜0.05).The pathological staining showed significantly reduction on renal interstitial fibrosis.The Notch/snail1 pathway,protein and mRNA expression of α-SMA were significant reduced with statistical significance (P＜0.01);protein and mRNA expression of E-Cad protein were significant increased with statistical significance (P＜0.01).It was concluded that TSN can protect the renal function of DN,delay the disease progression of DN,and inhibit epithelial-mesenchymal transdifferentiation (EMT) of renal tubular epithelial cells and renal interstitial fibrosis.Furthermore,the inhibition on EMT may be through the regulation of the Notch/snail1 pathway.
ABSTRACT
Objective:To observe the effect of Tangshenning on renal function and LOX-1 mRNA expression in the kidney of early diabetic nephropathy rats model. Methods:Early diabetic nephropathy rats model was made with high fat diet, STZ and unilateral ne-phregtomy. After the 12-week drug intervention, the rats were sacrificed, the kidneys were removed and the mRNA expressions of LOX-1 in the kidney were observed by the method of RT-PCR. Results:Early diabetic nephropathy rats model was successfully made by the triple-modeling method. The mRNA expression of LOX-1 in the kidney was significantly increased, and Tangshenning could low-er the expression of LOX-1 in the kidney in a dose-dependent manner. Conclusion:The abnormal expression of LOX-1 could be one of the mechanisms for diabetic nephropathy. Tangshening has good kidney protective effect through decreasing LOX-1 abnormal expression in the kidneys, which deserves further research.