中国中西医结合肾病杂志
中國中西醫結閤腎病雜誌
중국중서의결합신병잡지
CHINESE JOURNAL OF INTEGRATED TRADITIONAL AND WESTERN NEPHROLOGY
2014年
5期
392-396
,共5页
赵敬%王颖超%赵宗江%张新雪%杨美娟
趙敬%王穎超%趙宗江%張新雪%楊美娟
조경%왕영초%조종강%장신설%양미연
也页%糖肾平%足细胞%TGF-β1 -Smad2/3-ILK%信号转导通路%转分化
也頁%糖腎平%足細胞%TGF-β1 -Smad2/3-ILK%信號轉導通路%轉分化
야혈%당신평%족세포%TGF-β1 -Smad2/3-ILK%신호전도통로%전분화
Tangshenping%Podocyte%TGF-β1 -Smad2/3-ILK%Signal transduction pathway%Epithelial-mesen-chymal transition
也页目的：探讨糖肾平对高糖环境下脂多糖( lipopolysaccharide,LPS)刺激足细胞上皮间质转分化的影响,并探讨其作用机制。方法：以体外培养大鼠肾小球足细胞为研究对象,以高糖(25 mmol/L)、LPS(1μg/mL)刺激足细胞建立模型,分为正常组、高糖组、高糖+LPS组、厄贝沙坦组、抑制剂组、糖肾平小、中、大剂量组。采用Western blotting及RT-PCR方法检测足细胞中转化生长因子-β1( transforming growth factor-β1,TGF-β1)、Smad2/3、整合素连接激酶( integrin-linked kinase, ILK)、CD2相关蛋白(CD2AP)、α-平滑肌肌动蛋白(α-Smooth muscle actin,α-SMA)的表达水平。结果：与正常组比较,高糖组和高糖+LPS组足细胞TGF-β1、ILK、α-SMA蛋白及其mRNA表达明显增加(P<0.01),P-Smad2/3蛋白及Smad2/3 mRNA表达明显增加(P<0.01),CD2AP蛋白及其mRNA表达明显减少(P<0.01)；与高糖+LPS组比较,厄贝沙坦组足细胞P-Smad2/3、α-SMA蛋白表达明显减少(P<0.01),TGF-β1蛋白表达减少(P<0.05),TGF-β1,Smad2/3,α-SMAmRNA表达明显减少(P<0.01),ILK mRNA表达减少(P<0.05),CD2AP蛋白及其mRNA表达明显增加(P<0.01)；糖肾平大、中、小各剂量组足细胞TGF-β1蛋白表达明显减少(P<0.01),糖肾平大剂量组足细胞TGF-β1 mRNA表达减少(P<0.05),小、中剂量组表达明显减少(P<0.01)；糖肾平小、中、大各剂量组足细胞P-Smad2/3蛋白及Smad2/3mRNA表达均明显减少(P<0.01)；糖肾平小、大剂量组足细胞ILK mRNA表达减少(P<0.05),中剂量组表达明显减少(P<0.01)；糖肾平大剂量组足细胞α-SMA蛋白及其mRNA表达减少(P<0.05)；糖肾平小、中、大各剂量组足细胞CD2AP蛋白及其mRNA表达均明显增加(P<0.01)。结论：糖肾平能够降低足细胞TGF-β1、ILK蛋白及mRNA表达,降低P-Smad2/3蛋白及Smad2/3 mRNA表达,升高足细胞标志物CD2AP蛋白及mRNA表达,降低间充质细胞标志物α-SMA蛋白及mRNA表达,通过抑制TGF-β1-Smad2/3-ILK信号通路的激活减少足细胞转分化,保护足细胞,可能是其防治糖尿病肾病的作用机制之一。
也頁目的：探討糖腎平對高糖環境下脂多糖( lipopolysaccharide,LPS)刺激足細胞上皮間質轉分化的影響,併探討其作用機製。方法：以體外培養大鼠腎小毬足細胞為研究對象,以高糖(25 mmol/L)、LPS(1μg/mL)刺激足細胞建立模型,分為正常組、高糖組、高糖+LPS組、阨貝沙坦組、抑製劑組、糖腎平小、中、大劑量組。採用Western blotting及RT-PCR方法檢測足細胞中轉化生長因子-β1( transforming growth factor-β1,TGF-β1)、Smad2/3、整閤素連接激酶( integrin-linked kinase, ILK)、CD2相關蛋白(CD2AP)、α-平滑肌肌動蛋白(α-Smooth muscle actin,α-SMA)的錶達水平。結果：與正常組比較,高糖組和高糖+LPS組足細胞TGF-β1、ILK、α-SMA蛋白及其mRNA錶達明顯增加(P<0.01),P-Smad2/3蛋白及Smad2/3 mRNA錶達明顯增加(P<0.01),CD2AP蛋白及其mRNA錶達明顯減少(P<0.01)；與高糖+LPS組比較,阨貝沙坦組足細胞P-Smad2/3、α-SMA蛋白錶達明顯減少(P<0.01),TGF-β1蛋白錶達減少(P<0.05),TGF-β1,Smad2/3,α-SMAmRNA錶達明顯減少(P<0.01),ILK mRNA錶達減少(P<0.05),CD2AP蛋白及其mRNA錶達明顯增加(P<0.01)；糖腎平大、中、小各劑量組足細胞TGF-β1蛋白錶達明顯減少(P<0.01),糖腎平大劑量組足細胞TGF-β1 mRNA錶達減少(P<0.05),小、中劑量組錶達明顯減少(P<0.01)；糖腎平小、中、大各劑量組足細胞P-Smad2/3蛋白及Smad2/3mRNA錶達均明顯減少(P<0.01)；糖腎平小、大劑量組足細胞ILK mRNA錶達減少(P<0.05),中劑量組錶達明顯減少(P<0.01)；糖腎平大劑量組足細胞α-SMA蛋白及其mRNA錶達減少(P<0.05)；糖腎平小、中、大各劑量組足細胞CD2AP蛋白及其mRNA錶達均明顯增加(P<0.01)。結論：糖腎平能夠降低足細胞TGF-β1、ILK蛋白及mRNA錶達,降低P-Smad2/3蛋白及Smad2/3 mRNA錶達,升高足細胞標誌物CD2AP蛋白及mRNA錶達,降低間充質細胞標誌物α-SMA蛋白及mRNA錶達,通過抑製TGF-β1-Smad2/3-ILK信號通路的激活減少足細胞轉分化,保護足細胞,可能是其防治糖尿病腎病的作用機製之一。
야혈목적：탐토당신평대고당배경하지다당( lipopolysaccharide,LPS)자격족세포상피간질전분화적영향,병탐토기작용궤제。방법：이체외배양대서신소구족세포위연구대상,이고당(25 mmol/L)、LPS(1μg/mL)자격족세포건립모형,분위정상조、고당조、고당+LPS조、액패사탄조、억제제조、당신평소、중、대제량조。채용Western blotting급RT-PCR방법검측족세포중전화생장인자-β1( transforming growth factor-β1,TGF-β1)、Smad2/3、정합소련접격매( integrin-linked kinase, ILK)、CD2상관단백(CD2AP)、α-평활기기동단백(α-Smooth muscle actin,α-SMA)적표체수평。결과：여정상조비교,고당조화고당+LPS조족세포TGF-β1、ILK、α-SMA단백급기mRNA표체명현증가(P<0.01),P-Smad2/3단백급Smad2/3 mRNA표체명현증가(P<0.01),CD2AP단백급기mRNA표체명현감소(P<0.01)；여고당+LPS조비교,액패사탄조족세포P-Smad2/3、α-SMA단백표체명현감소(P<0.01),TGF-β1단백표체감소(P<0.05),TGF-β1,Smad2/3,α-SMAmRNA표체명현감소(P<0.01),ILK mRNA표체감소(P<0.05),CD2AP단백급기mRNA표체명현증가(P<0.01)；당신평대、중、소각제량조족세포TGF-β1단백표체명현감소(P<0.01),당신평대제량조족세포TGF-β1 mRNA표체감소(P<0.05),소、중제량조표체명현감소(P<0.01)；당신평소、중、대각제량조족세포P-Smad2/3단백급Smad2/3mRNA표체균명현감소(P<0.01)；당신평소、대제량조족세포ILK mRNA표체감소(P<0.05),중제량조표체명현감소(P<0.01)；당신평대제량조족세포α-SMA단백급기mRNA표체감소(P<0.05)；당신평소、중、대각제량조족세포CD2AP단백급기mRNA표체균명현증가(P<0.01)。결론：당신평능구강저족세포TGF-β1、ILK단백급mRNA표체,강저P-Smad2/3단백급Smad2/3 mRNA표체,승고족세포표지물CD2AP단백급mRNA표체,강저간충질세포표지물α-SMA단백급mRNA표체,통과억제TGF-β1-Smad2/3-ILK신호통로적격활감소족세포전분화,보호족세포,가능시기방치당뇨병신병적작용궤제지일。
Objective:To observe the influence of the capsule of Tangshenping on LPS-stimulated podocytes epithelial-mesenchymal transition ( EMT) under high glucose conditions,and explore its possible mechanism. Methods:Using cultured rat glo-merular podocytes as the object of study, making model with high glucose(25 mmol/L)、LPS(1 μg/ml) stimulating podocytes, and divide them into 8 groups:control group, high glucose group, high glucose plus LPS group, irbesartan group, low-dose group of Tan-gshenping capsule, moderate dose group of Tangshenping capsule, high-dose group of Tangshenping, inhibitor group. Detect podo-cyte transforming growth factor-β1(TGF-β1), Smad2/3, integrin-linked kinase(ILK), CD2AP and α-smooth muscle actin (α-SMA) mRNA expression of each group by Western blotting and RT-PCR. Results:Compared with the control group, the ex-pression of podocyte TGF-β1、ILK、α-SMA protein and mRNA increased definitely(P<0. 01), P-Smad2/3 protein and Smad2/3 mRNA expression increased definitely (P<0. 01), CD2AP protein and mRNA decreased definitely(P<0. 01) in the high glucose group and high glucose plus LPS group. Compared with the high glucose plus LPS group, the expression of podocyte P-Smad2/3、α-SMA protein decreased definitely(P<0. 01), TGF-β1 protein decreased (P<0. 05), TGF-β1,Smad2/3,α-SMAmRNA de-creased definitely(P<0. 01), ILKmRNA decreased(P<0. 05), CD2AP protein and mRNA increased definitely(P<0. 01) in the irbesartan group;The expression of podocyte TGF-β1 protein decreased definitely(P<0. 01) in the low, moderate and high dose group of Tangshenping, TGF-β1 mRNA decreased in the high dose group of Tangshenping(P<0. 05)and decreased definitely in the moderate,low dose group of Tangshenping (P<0. 01); In the low, moderate and high dose group of Tangshenping, podocyte P-Smad2/3 protein and Smad2/3mRNA expression decreased definitely(P<0. 01);The expression of podocyte ILKmRNA decreased in the low and high dose group of Tangshenping(P<0. 05), decreased definitely in the moderate dose group(P<0. 01); In the high dose group of Tangshenping, podocyte α-SMA protein and mRNA expression decreased (P<0. 05);In the low,moderate and high dose group of Tangshenping, podocyte CD2AP protein and mRNA expression increased definitely(P<0. 01). Conclusion:Tangshenp-ing can reduce podocyte TGF-β1, ILK protein and mRNA expression, reduce P-Smad2/3 protein and Smad2/3mRNA expression, increase podocytes marker CD2AP protein and mRNA expression and reduce mesenchymal cell marker α-SMA protein and mRNA expression, Tangshenping can relieve podocytes epithelial-mesenchymal transition by inhibiting TGF-β1 -Smad2/3 -ILK signal transduction pathway, this mechanism may be considered as a means of prevention and treatment for diabetic nephropathy.
