浙江医学
浙江醫學
절강의학
ZHEJIANG MEDICAL JOURNAL
2013年
19期
1718-1721
,共4页
TGF-β1%足细胞%凋亡
TGF-β1%足細胞%凋亡
TGF-β1%족세포%조망
TGF- β1%Podocyte%Apoptosis
目的:探讨转化生长因子-β1(TGF-β1)是否能诱导小鼠足细胞株凋亡以及诱导凋亡的途径。方法(1)体外培养小鼠足细胞株;(2)应用不同浓度(10-1~104ng/L)TGF-β1诱导小鼠足细胞凋亡;(3)103ng/L TGF-β1诱导不同时间(12、24、48h)后观察足细胞凋亡情况;(4)采用流式细胞仪检测Annexin V、Caspase 3;(5)实时定量PCR及Western印迹法检测p38丝裂原活化蛋白激酶(p38MAPK)、Smad2、Smad3、Smad7mRNA及蛋白质的表达情况。结果(1)小鼠足细胞在不同浓度(10-1~104ng/L) TGF-β1刺激下,细胞凋亡率明显高于正常对照组(均P<0.05);(2)随着TGF-β1浓度增加、凋亡时间增加,足细胞凋亡率增高(P<0.05);p38MAPK、Smad2、Smad3、Smad7 mRNA表达增加;TGF-β1103ng/L为最佳诱导小鼠足细胞凋亡浓度,24h为最佳诱导凋亡时间;(3)与正常对照组比较,103ng/LTGF-β1诱导后Caspase 3阳性细胞比率显著增加(P<0.01),p38MAPK、Smad2、Smad3、Smad7蛋白质表达亦显著增加(P<0.01)。结论 TGF-β1能诱导小鼠足细胞凋亡,呈浓度及时间依赖性。TGF-β1通过Smad通路、p38MAPK通路以及线粒体通路诱导小鼠足细胞凋亡。
目的:探討轉化生長因子-β1(TGF-β1)是否能誘導小鼠足細胞株凋亡以及誘導凋亡的途徑。方法(1)體外培養小鼠足細胞株;(2)應用不同濃度(10-1~104ng/L)TGF-β1誘導小鼠足細胞凋亡;(3)103ng/L TGF-β1誘導不同時間(12、24、48h)後觀察足細胞凋亡情況;(4)採用流式細胞儀檢測Annexin V、Caspase 3;(5)實時定量PCR及Western印跡法檢測p38絲裂原活化蛋白激酶(p38MAPK)、Smad2、Smad3、Smad7mRNA及蛋白質的錶達情況。結果(1)小鼠足細胞在不同濃度(10-1~104ng/L) TGF-β1刺激下,細胞凋亡率明顯高于正常對照組(均P<0.05);(2)隨著TGF-β1濃度增加、凋亡時間增加,足細胞凋亡率增高(P<0.05);p38MAPK、Smad2、Smad3、Smad7 mRNA錶達增加;TGF-β1103ng/L為最佳誘導小鼠足細胞凋亡濃度,24h為最佳誘導凋亡時間;(3)與正常對照組比較,103ng/LTGF-β1誘導後Caspase 3暘性細胞比率顯著增加(P<0.01),p38MAPK、Smad2、Smad3、Smad7蛋白質錶達亦顯著增加(P<0.01)。結論 TGF-β1能誘導小鼠足細胞凋亡,呈濃度及時間依賴性。TGF-β1通過Smad通路、p38MAPK通路以及線粒體通路誘導小鼠足細胞凋亡。
목적:탐토전화생장인자-β1(TGF-β1)시부능유도소서족세포주조망이급유도조망적도경。방법(1)체외배양소서족세포주;(2)응용불동농도(10-1~104ng/L)TGF-β1유도소서족세포조망;(3)103ng/L TGF-β1유도불동시간(12、24、48h)후관찰족세포조망정황;(4)채용류식세포의검측Annexin V、Caspase 3;(5)실시정량PCR급Western인적법검측p38사렬원활화단백격매(p38MAPK)、Smad2、Smad3、Smad7mRNA급단백질적표체정황。결과(1)소서족세포재불동농도(10-1~104ng/L) TGF-β1자격하,세포조망솔명현고우정상대조조(균P<0.05);(2)수착TGF-β1농도증가、조망시간증가,족세포조망솔증고(P<0.05);p38MAPK、Smad2、Smad3、Smad7 mRNA표체증가;TGF-β1103ng/L위최가유도소서족세포조망농도,24h위최가유도조망시간;(3)여정상대조조비교,103ng/LTGF-β1유도후Caspase 3양성세포비솔현저증가(P<0.01),p38MAPK、Smad2、Smad3、Smad7단백질표체역현저증가(P<0.01)。결론 TGF-β1능유도소서족세포조망,정농도급시간의뢰성。TGF-β1통과Smad통로、p38MAPK통로이급선립체통로유도소서족세포조망。
Objective To investigate the effect of TGF- β1 on apoptosis of mouse podocytes and its signal pathway. Methods Cultured mouse podocytes were treated with different concentrations of TGF- β1(0,1x10-1~104 ng/L) for 24 h, or treated with 103 ng/L TGF- β1 for different time duration (12, 24, 48h). Annexin V and Caspase 3 were measured by flow cytometry. The mRNAs and proteins of P38MAPK, Smad2, Smad3 and Smad7 were detected by real time PCR and Western blotting. Results Compared to controls(0ng/L TGF- β1) the ratio of apoptotic podocytes and mRNA expressions of p38MAPK, Smad2, Smad3 and Smad7 were increased in TGF- β1 (10-1~104ng/L) treatment groups in a concentration and time- dependent manner (P<0.05). Treatment with TGF- β1 (103ng/L) for 24 h induced maximal apoptosis in podocytes. The ratio of Caspase3- positive podocytes was increased after treated with TGF- β1(103ng/L, 24h) compared to controls(P<0.01). Conclusion TGF- β1 induces the apop-tosis of mouse podocytes time- dependently and dose- dependently, in which Smad, p38MAPK signal pathway and mitochondri-on might be involved.