中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2013年
8期
606-609
,共4页
苗宏志%李波%徐斐翔%金莉%王国忠%林岩%邓志会%肖薇%李光伟
苗宏誌%李波%徐斐翔%金莉%王國忠%林巖%鄧誌會%肖薇%李光偉
묘굉지%리파%서비상%금리%왕국충%림암%산지회%초미%리광위
受体,钙敏感%肌细胞,平滑肌%肺动脉%衔接蛋白质类,信号转导
受體,鈣敏感%肌細胞,平滑肌%肺動脈%銜接蛋白質類,信號轉導
수체,개민감%기세포,평활기%폐동맥%함접단백질류,신호전도
Receptors,calcium-sensing%Myocytes,smooth muscle%Pulmonary artery%Adaptor protcins,signal transducing
目的 探讨钙敏感受体在缺氧诱导的大鼠肺动脉平滑肌细胞(PAMSCs)增殖中的信号转导途径及其作用.方法 Ⅱ型胶原酶消化法提取、培养大鼠PAMSCs.通过缺氧培养箱内(93％N2,2％02,5％ CO2)培养24 h的方法复制细胞缺氧模型.应用Western印迹技术分析不同处理情况下增殖细胞核抗原(PCNA)及磷酸化的细胞外调解蛋白激酶1,2(p-ERK1,2)蛋白在PAMSCs的相对表达量；采用流式细胞技术检测不同处理因素对细胞增殖周期及增殖指数的影响,应用5-溴脱氧尿嘧啶核苷(BrdU)掺入法分析不同处理因素对细胞DNA合成的影响.结果 缺氧引起PAMSCs的PCNA(0.528±0.028)及p-ERK1,2(1.12±0.05、0.91±0.06)表达水平上调,均显著高于对照组的0.243±0.025及0.47±0.03、0.40±0.03(均P＜0.05),BrdU掺入量(143.3±4.2)和细胞增殖指数(12.5±0.9)也显著高于对照组的(100.0±5.4和7.5±1.2)(均P＜0.05)；钙敏感受体激动剂GdCl3能够放大缺氧的上述作用(0.770±0.039,1.50±0.06,1.61 ±0.05,187.4 ±3.9,19.8±0.6)(与对缺氧组比较,均P ＜0.05),但此效应可以被ERK1,2通路阻断剂PD98059抑制(0.441±0.020,0.71±0.07,0.72±0.06,115.5±4.0,9.3±1.1)(与缺氧+GdCl3组比较,均P＜0.05).结论 CaSR通过ERK1,2信号通路参与缺氧诱导的大鼠PAMSCs增殖.
目的 探討鈣敏感受體在缺氧誘導的大鼠肺動脈平滑肌細胞(PAMSCs)增殖中的信號轉導途徑及其作用.方法 Ⅱ型膠原酶消化法提取、培養大鼠PAMSCs.通過缺氧培養箱內(93％N2,2％02,5％ CO2)培養24 h的方法複製細胞缺氧模型.應用Western印跡技術分析不同處理情況下增殖細胞覈抗原(PCNA)及燐痠化的細胞外調解蛋白激酶1,2(p-ERK1,2)蛋白在PAMSCs的相對錶達量；採用流式細胞技術檢測不同處理因素對細胞增殖週期及增殖指數的影響,應用5-溴脫氧尿嘧啶覈苷(BrdU)摻入法分析不同處理因素對細胞DNA閤成的影響.結果 缺氧引起PAMSCs的PCNA(0.528±0.028)及p-ERK1,2(1.12±0.05、0.91±0.06)錶達水平上調,均顯著高于對照組的0.243±0.025及0.47±0.03、0.40±0.03(均P＜0.05),BrdU摻入量(143.3±4.2)和細胞增殖指數(12.5±0.9)也顯著高于對照組的(100.0±5.4和7.5±1.2)(均P＜0.05)；鈣敏感受體激動劑GdCl3能夠放大缺氧的上述作用(0.770±0.039,1.50±0.06,1.61 ±0.05,187.4 ±3.9,19.8±0.6)(與對缺氧組比較,均P ＜0.05),但此效應可以被ERK1,2通路阻斷劑PD98059抑製(0.441±0.020,0.71±0.07,0.72±0.06,115.5±4.0,9.3±1.1)(與缺氧+GdCl3組比較,均P＜0.05).結論 CaSR通過ERK1,2信號通路參與缺氧誘導的大鼠PAMSCs增殖.
목적 탐토개민감수체재결양유도적대서폐동맥평활기세포(PAMSCs)증식중적신호전도도경급기작용.방법 Ⅱ형효원매소화법제취、배양대서PAMSCs.통과결양배양상내(93％N2,2％02,5％ CO2)배양24 h적방법복제세포결양모형.응용Western인적기술분석불동처리정황하증식세포핵항원(PCNA)급린산화적세포외조해단백격매1,2(p-ERK1,2)단백재PAMSCs적상대표체량；채용류식세포기술검측불동처리인소대세포증식주기급증식지수적영향,응용5-추탈양뇨밀정핵감(BrdU)참입법분석불동처리인소대세포DNA합성적영향.결과 결양인기PAMSCs적PCNA(0.528±0.028)급p-ERK1,2(1.12±0.05、0.91±0.06)표체수평상조,균현저고우대조조적0.243±0.025급0.47±0.03、0.40±0.03(균P＜0.05),BrdU참입량(143.3±4.2)화세포증식지수(12.5±0.9)야현저고우대조조적(100.0±5.4화7.5±1.2)(균P＜0.05)；개민감수체격동제GdCl3능구방대결양적상술작용(0.770±0.039,1.50±0.06,1.61 ±0.05,187.4 ±3.9,19.8±0.6)(여대결양조비교,균P ＜0.05),단차효응가이피ERK1,2통로조단제PD98059억제(0.441±0.020,0.71±0.07,0.72±0.06,115.5±4.0,9.3±1.1)(여결양+GdCl3조비교,균P＜0.05).결론 CaSR통과ERK1,2신호통로삼여결양유도적대서PAMSCs증식.
Objective To explore the cell signal transduction pathway of calcium-sensing receptor (CaSR) mediated hypoxia-induced proliferation of rat pulmonary artery smooth muscle cells (PASMCs).Methods The expressions of proliferating cell nuclear antigen (PCNA) and extracellular signal-regulated protein kinase 1,2 (ERK1,2) were analyzed by Western blot.Cell proliferation was tested by a 5-bromo2-deoxyuridine(BrdU) incorporation assay.Cell cycle and proliferation index (PI) were analyzed by flow cytometry.Results Hypoxia significantly increased the expression of PCNA (0.528 ±0.028),p-ERK1,2(1.12 ±0.05,0.91 ±0.06),BrdU incorporation (143.3 ±4.2) and cell proliferation index (12.5 ±0.9)(all P ＜0.05,versus control group,0.243 ±0.025,0.47 ±0.03,0.40 ±0.03,100.0 ±5.4,7.5 ±1.2).Gadolinium chloride (GdCl3,a CaSR agonist) amplified the effect of hypoxia (0.770 ± 0.039,1.50±0.06,1.61 ±0.05,187.4 ±3.9,19.8 ±0.6,all P ＜0.05).PD98059 (a MEK1 inhibitor)decreased the up-regulation of PCNA expression,BrdU incorporation and the increase of cell proliferation index induced by hypoxia and GdCl3 in PASMCs (0.441 ± 0.020,0.71 ± 0.07,0.72 ± 0.06,115.5 ±4.0,9.3 ± 1.1,all P ＜ 0.05).Conclusion Calcium-sensing receptor mediates hypoxia-induced proliferation of rat pulmonary artery smooth muscle cells through ERK1,2 pathways.
