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Objective:To investigate the renal protective effect of Tangshenping capsule (Tangshenping) on diabetic nephropathy (DN) KKAy mice and its effect on Wnt/β-catenin signaling pathway. Method:Sixty female Sprague-Dawley KKAy mice aged 10 weeks old were induced with KKAy rat feed for 10 weeks. The DN animal model was successfully determined with blood glucose (>16.7 mmol·L-1) and 24 hour urine protein (>0.4 mg). The model mice were randomly divided into a model group, an irbesartan group, and low, medium and high-dose Tangshenping group, with 10 female C57BL/6J mice as a control group. The treatment groups were given the corresponding drugs by gavage. The normal group and the model group were given an equal volume of deionized water by gavage. The intragastric dose was 0.01 mL·g-1 body weight coefficient once a day. The general conditions of the mice were observed, the body mass was weighed every 4 weeks, and 24 h urine protein was quantified. At the 26th week, the blood was collected from eyeballs, and the mice were put to death. The quality of the kidneys, serum blood urea nitrogen (BUN), serum creatinine (SCr), triglyceride (TG), malondialdehyde (MDA), nitric oxide (NO) and superoxide dismutase (SOD) content were measured. In situ hybridization and immunohistochemistry were used to detect the expressions of Wnt4, glycogen synthase kinase 3β(GSK3β) and β-catenin in kidney tissues. Result:Compared with model group, body mass, kidney mass/body mass, and 24 h urine protein were significantly lower in high-dose Tangshenping group (PPPβ and β-catenin were decreased (PConclusion:Tangshenping may inhibit the activation of Wnt/β-catenin signaling pathway, reverse the transdifferentiation of renal tubular epithelial cells in DN KKAy mice, delay the progression of renal interstitial fibrosis, and then exert renal protection.
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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.
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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.
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To study the effect of the Tangshenping containing serum on the proliferation of the high glucose-induced epithelial cells of renal tubules.To prepare durg-contained serum from rats to enter into in vitor reaction system,the cellscultured via 10％ FBS-RPMI 1640 were randomly divided into 7groups:the normal group,themodel group,the Y27632 group,theirbesartangroup,the small dose Tangshenping group,the medium dose Tangshenping group and the high dose Tangshenping group and The cells were cultured in 3000 cells/well and grown in 96-well plates.Each group had 8 wells,then detect the effects of all serum sections on the proliferation of high glucose-induced epithelial cells of kidney tubules by the MTT colorimetric method after cultured for 12 h,24 h,48 h,and 60 h.Based on the results above,cell protein were extracted from each group at 24 h,and the expression of RhoA,ROCK1,α-SMA and E-cadherin in each group were detected by Western blotting.After high glucose stimulation,the shape of cell was shuttle-like or irregular triangle,the way it grew was radial;after the intervention of the corresponding serum,the shape of the cell was fiat and irregular polygonal.Started with 12h,compared with the normal group,OD value of other groups increased;at the 24h、48hand 60h,compared with the normal group,OD value of high glucose groupincreased significantly (P＜0.01);compared with the high glucose group,OD value of treatment groups decreased (P＜0.05);and 48 h,compared with the Y27632group,irbesartan groupand Tangshenping high dose group,OD value of Tangshenping low and medium dose groups decreased (P＜0.05);60 h,compared with Y27632 group,OD value of Tangshenping medium dose groups decreased;compared with irbesartan group,OD value of o Tangshenpinggroupsdecreased (P＜0.05);compared with Tangshenping high dose group,OD value of Tangshenpinglow groupsdecreased (P＜0.05) Western blotting analysis showed that compared with normal group,the expression of E-Cadherin protein in high glucose group reduced,and the expression of RhoA,ROCK1 and α-SMA protein increased;compared with high glucose group,the expression of E-Cadherin protein in each treating group increased,and the Tangshenping large dose group wassignificantly different (P＜0.01);the expression of RhoA,ROCK1 and or-SMA protein reduced,Tangshenping,the large dose group was significantlydifferent (P＜0.01);Compared with the Y27632 group,the expression of E-cadherin,ROCK1 and α-SMA protein in Tangshenping large dose group had no significant difference,while the expression of RhoAproteinreduced (P ＜0.01).Compared with theirbesartan group,the expression of E-cadherin,RhoA,ROCK1 and α-SMA protein in Tangshenping large dose group had no significant difference (P＞0.05).The Tangshenping containing serum is abletoinhibit the proliferation of high glucose-induced epithelial cells of kidney tubules,and can reverse renal tubular-epithelial cell transdifferentiation via regulating RhoA/ROCK signaling pathway,and restrain renal interstitial fibrosis,thereby delaying the pathogenesis of diabetic kidney disease.
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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.
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Diabetic kidney disease (DKD) is a serious long-term complication and major death cause of patients who have suffered from diabetes mellitus (DM),which is a serious threat to the health of human being.Professor Zhao Zongjiang first came up with the of DKD "consumptive kidney disease" scientific theory,and led the scientific research team for DKD research.Our team had achieved some results,which were mainly focused on the animal experiments and cell culture experiment.And the research fields covered pharmacodynamics,oxidative stress levels and signal transduction pathways.Focusing on the reduction of extracellular matrix in the glomerular mesangial area and tubulointerstitial deposition,inhibition of podocyte injury,inhibition of podocyte apoptosis,inhibition and reversal of podocyte transdifferentiation,inhibition of renal tubular epithelial cell transdifferentiation and other scientific issues,this paper systematically explored the biological mechanism of DKD "consumptive kidney disease" scientific theory.This paper explained how "consumptive kidney disease" scientific theory was proposed and summarized related research results of our research team and developed traditional Chinese medicine (TCM) theory by supplying the TCM "consumptive organ disease" theory.
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Diabetic nephropathy was one of the diseases with high incidence and difficult to treat in modern society. Modern medicine had focused on the prevention and treatment of this difficult disease to delay its development. ProfessorZhao Zongjiang, who had been working on the treatment of kidney diseases with traditional Chinese medicine (TCM) and modern medicine for many years, had deep understanding on integrative medicine treatment of chronic kidney diseases (CKD), diabetic nephropathy treatment with unique TCM principles, methods, prescriptions and herbs, with rich clinical experiences, rational medication and significant therapeutic effects. Under the guidance of professorZhao Zongjiang, this article started with consumptive kidney disease in combination with a large amount of professorZhao Zongjiang’s research results, for the discussion of professor Zhao Zongjiang’s academic thoughts and clinical experiences in disease treatment to provide references in this field.