中华肾脏病杂志
中華腎髒病雜誌
중화신장병잡지
2014年
10期
763-769
,共7页
郭云珊%张爱平%王艳侠%张磊%李宏栋%李震%丁尧海
郭雲珊%張愛平%王豔俠%張磊%李宏棟%李震%丁堯海
곽운산%장애평%왕염협%장뢰%리굉동%리진%정요해
β连环素%纤维化%甲状旁腺素%间充质转分化%肾小管上皮细胞
β連環素%纖維化%甲狀徬腺素%間充質轉分化%腎小管上皮細胞
β련배소%섬유화%갑상방선소%간충질전분화%신소관상피세포
Beta Catenin%Fibrosis%Parathyroid hormone%Epithelial to mesenchymal transition%Renal tubular epithelial cells
目的 观察甲状旁腺素(PTH)对人肾小管上皮细胞转分化的影响,并探讨β连环素(β-catenin)信号通路在此过程中的作用.方法 应用实时定量PCR、Western印迹、免疫荧光等方法,观察PTH诱导人近曲小管上皮细胞(HK-2细胞)α平滑肌肌动蛋白(α-SMA)、E钙黏蛋白(E-cadherin)和β-catenin的表达情况.采用siRNA阻断β-catenin信号通路以明确PTH发挥作用的信号途径.结果 10-10 mol/LPTH刺激HK-2细胞48 h时α-SMA mRNA和蛋白表达显著增加(P＜ 0.01),E-cadherin表达下降(P＜0.01).10-10 mol/L PTH作用HK-2细胞48 h后,免疫荧光检测结果显示细胞胞质中α-SMA表达增多,E-cadherin表达减少.PTH可激活β-catenin信号通路;β-catenin siRNA干预细胞后,α-SMA mRNA和蛋白表达均下调(P＜0.05),而E-cadhenn表达上调(P＜0.05).结论 PTH可诱导HK-2细胞间充质转分化,其作用可能是通过β-catenin信号通路来实现的.
目的 觀察甲狀徬腺素(PTH)對人腎小管上皮細胞轉分化的影響,併探討β連環素(β-catenin)信號通路在此過程中的作用.方法 應用實時定量PCR、Western印跡、免疫熒光等方法,觀察PTH誘導人近麯小管上皮細胞(HK-2細胞)α平滑肌肌動蛋白(α-SMA)、E鈣黏蛋白(E-cadherin)和β-catenin的錶達情況.採用siRNA阻斷β-catenin信號通路以明確PTH髮揮作用的信號途徑.結果 10-10 mol/LPTH刺激HK-2細胞48 h時α-SMA mRNA和蛋白錶達顯著增加(P＜ 0.01),E-cadherin錶達下降(P＜0.01).10-10 mol/L PTH作用HK-2細胞48 h後,免疫熒光檢測結果顯示細胞胞質中α-SMA錶達增多,E-cadherin錶達減少.PTH可激活β-catenin信號通路;β-catenin siRNA榦預細胞後,α-SMA mRNA和蛋白錶達均下調(P＜0.05),而E-cadhenn錶達上調(P＜0.05).結論 PTH可誘導HK-2細胞間充質轉分化,其作用可能是通過β-catenin信號通路來實現的.
목적 관찰갑상방선소(PTH)대인신소관상피세포전분화적영향,병탐토β련배소(β-catenin)신호통로재차과정중적작용.방법 응용실시정량PCR、Western인적、면역형광등방법,관찰PTH유도인근곡소관상피세포(HK-2세포)α평활기기동단백(α-SMA)、E개점단백(E-cadherin)화β-catenin적표체정황.채용siRNA조단β-catenin신호통로이명학PTH발휘작용적신호도경.결과 10-10 mol/LPTH자격HK-2세포48 h시α-SMA mRNA화단백표체현저증가(P＜ 0.01),E-cadherin표체하강(P＜0.01).10-10 mol/L PTH작용HK-2세포48 h후,면역형광검측결과현시세포포질중α-SMA표체증다,E-cadherin표체감소.PTH가격활β-catenin신호통로;β-catenin siRNA간예세포후,α-SMA mRNA화단백표체균하조(P＜0.05),이E-cadhenn표체상조(P＜0.05).결론 PTH가유도HK-2세포간충질전분화,기작용가능시통과β-catenin신호통로래실현적.
Objective To investigate the effect of parathyroid hormone (PTH) on the epithelial to mesenchymal transition (EMT) in human renal proximal tubular epithelial cells (HK-2 cells),and determine the role of β-catenin signaling pathway.Method The expression of α-smooth muscle actin (α-SMA),E-cadherin and β-catenin in HK-2 cells was measured by real-time PCR,Western blotting and immunofluorescence technique.The signaling pathway by which PTH activated EMT in HK -2 cells was identified by using synthetic β-catenin siRNA.Results Parathyroid hormone (10-10mol/ L) increased α-SMA expression and decreased E-cadherin expression in HK-2 cells (P＜ 0.01,respectively).Untreated cells showed the expression of E-cadherin,whereas α-SMA staining was noticeably increased in cells treated with PTH.β-catenin activity was significantly increased after exposed to PTH.Theα-SMA expression was decreased strongly and E-cadherin expression was increased after β-catenin siRNA transfection (all P ＜ 0.05).Conclusion PTH significantly induces epithelial to mesenchymal transition in HK-2 cells throughβ-catenin signaling pathway.
