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
This study was aimed to explore the renoprotective effects of Tang-Shen-Ping (TSP) on RhoA/ROCK signaling pathway in KKAy mice with diabetic kidney disease (DKD).A total of 60 female 10-week SPF degree KKAy mice,which were fed with KK special food for 10 weeks,were made into DKD model.Mice were randomly divided in the model group,irbesartan group,low-,medium-and high-dose TSP group (0.525 g· kg-1,1.05 g· kg-1,and 2.1 g· kg-1).Ten female C57BL/6J mice were used as the normal control group.Mice of each group were intragastrically administered with corresponding medicine,respectively,while mice of the control group and the model group were given deionized water of the equal volume.The body weight was measured and the 24-hour urine protein quantification was detected every 4 weeks.At the end of the 26th week,all mice were sacrificed and the biochemical indicators,such as fasting blood glucose (FBG),serum blood urea nitrogen (BUN),serum creatinine (Scr),and triglyceride (TG) were measured.HE staining,Mallory staining and PAS staining were used to observe the pathological morphology of kidney tissues.Immunohistochemistry (IHC) and in situ hybridization (ISH) were used in the detection of transforming growth factor-β1 (TGF-β1),Ras homolog gene family member A (RhoA),Rho-associated coiled-coil-containing protein kinase 1 (ROCK1),α-smooth muscle actin (α-SMA),E-Cadherin (E-Cad) mRNA and protein expression.The results showed that compared with the model group,there were significant differences on body weight,the ratio of kidney weight to body weight,and urinary protein in the middle-and high-dose TSP group (P ＜ 0.01);the renal pathological damage was obviously decreased;contents of FBG,BUN,Scr and TG decreased (P ＜ 0.01);mRNA and protein expression of E-Cadherin increased;mRNA and protein expression of TGF-β1,RhoA,ROCK1 and α-SMA decreased with significant difference in the middle-and high-dosc TSP group (P ＜ 0.01).It was concluded that the renoprotective effects and epithelial-mesenchymal transdifferentiation (EMT) of renal tubular epithelial cells of TSP on DKD KKAy mice may be related to the regulation of RhoA/ROCK signaling pathway.