中华劳动卫生职业病杂志
中華勞動衛生職業病雜誌
중화노동위생직업병잡지
CHINESE JOURNAL OF INDUSTRIAL HYGIENE AND OCCUPATIONAL DISEASES
2011年
10期
731-734
,共4页
陈婵%王志翊%王良兴%杜晓红%赵晓微
陳嬋%王誌翊%王良興%杜曉紅%趙曉微
진선%왕지익%왕량흥%두효홍%조효미
葛根素%低氧%肌,平滑,血管
葛根素%低氧%肌,平滑,血管
갈근소%저양%기,평활,혈관
PUERARIN%Anoxia%Muscle,smooth,vascular
目的 观察葛根素对低氧性肺动脉平滑肌细胞( PASMCs)增殖与凋亡及电压门控型钾离子通道亚型(Kv1.5)表达的影响,探讨葛根素在改善低氧性肺动脉高压与抑制肺血管重建中可能的作用机制.方法 原代培养大鼠PASMCs,随机分为正常对照组(5％CO2常氧),低氧组(5％O2、5％CO2、90％N2三气培养),3个浓度葛根素干预组(在低氧组基础上分别加入终浓度为1×104、1×10-4、1×10-3 mol/L葛根素,37℃培养24 h).采用CCK-8法和流式细胞仪检测细胞增殖情况,分光光度法检测caspase-3活力,蛋白印迹及实时定量聚合酶链反应(PCR)法分别检测Kv1.5蛋白和mRNA表达.结果 与正常对照组(细胞活性:0.940±0.045,S期细胞比例:9.67％±1.28％,Caspase-3活力:0.1073±0.0113,Kv 1.5蛋白:0.886±0.038,Kv 1.5 mRNA 0.0377±0.0031)比较,低氧组细胞活性(1.296±0.034)、S期细胞比例(18.19％±1.19％)升高,Caspase-3活力(0.0664±0.0049)下降,Kv1.5蛋白(0.602±0.064)及mRNA (0.0108±0.0014)表达下降,差异均有统计学意义(P＜0.05)；与低氧组比较,各剂量葛根素干预组细胞活性和S期细胞比例下降,Caspase-3活力升高,Kv 1.5蛋白及mRNA表达上调,差异均有统计学意义(P＜0.05).结论 葛根素可抑制低氧性PASMCs增殖,促进其凋亡,其作用机制可能通过上调Kv 1.5表达抑制PASMCs的增殖.
目的 觀察葛根素對低氧性肺動脈平滑肌細胞( PASMCs)增殖與凋亡及電壓門控型鉀離子通道亞型(Kv1.5)錶達的影響,探討葛根素在改善低氧性肺動脈高壓與抑製肺血管重建中可能的作用機製.方法 原代培養大鼠PASMCs,隨機分為正常對照組(5％CO2常氧),低氧組(5％O2、5％CO2、90％N2三氣培養),3箇濃度葛根素榦預組(在低氧組基礎上分彆加入終濃度為1×104、1×10-4、1×10-3 mol/L葛根素,37℃培養24 h).採用CCK-8法和流式細胞儀檢測細胞增殖情況,分光光度法檢測caspase-3活力,蛋白印跡及實時定量聚閤酶鏈反應(PCR)法分彆檢測Kv1.5蛋白和mRNA錶達.結果 與正常對照組(細胞活性:0.940±0.045,S期細胞比例:9.67％±1.28％,Caspase-3活力:0.1073±0.0113,Kv 1.5蛋白:0.886±0.038,Kv 1.5 mRNA 0.0377±0.0031)比較,低氧組細胞活性(1.296±0.034)、S期細胞比例(18.19％±1.19％)升高,Caspase-3活力(0.0664±0.0049)下降,Kv1.5蛋白(0.602±0.064)及mRNA (0.0108±0.0014)錶達下降,差異均有統計學意義(P＜0.05)；與低氧組比較,各劑量葛根素榦預組細胞活性和S期細胞比例下降,Caspase-3活力升高,Kv 1.5蛋白及mRNA錶達上調,差異均有統計學意義(P＜0.05).結論 葛根素可抑製低氧性PASMCs增殖,促進其凋亡,其作用機製可能通過上調Kv 1.5錶達抑製PASMCs的增殖.
목적 관찰갈근소대저양성폐동맥평활기세포( PASMCs)증식여조망급전압문공형갑리자통도아형(Kv1.5)표체적영향,탐토갈근소재개선저양성폐동맥고압여억제폐혈관중건중가능적작용궤제.방법 원대배양대서PASMCs,수궤분위정상대조조(5％CO2상양),저양조(5％O2、5％CO2、90％N2삼기배양),3개농도갈근소간예조(재저양조기출상분별가입종농도위1×104、1×10-4、1×10-3 mol/L갈근소,37℃배양24 h).채용CCK-8법화류식세포의검측세포증식정황,분광광도법검측caspase-3활력,단백인적급실시정량취합매련반응(PCR)법분별검측Kv1.5단백화mRNA표체.결과 여정상대조조(세포활성:0.940±0.045,S기세포비례:9.67％±1.28％,Caspase-3활력:0.1073±0.0113,Kv 1.5단백:0.886±0.038,Kv 1.5 mRNA 0.0377±0.0031)비교,저양조세포활성(1.296±0.034)、S기세포비례(18.19％±1.19％)승고,Caspase-3활력(0.0664±0.0049)하강,Kv1.5단백(0.602±0.064)급mRNA (0.0108±0.0014)표체하강,차이균유통계학의의(P＜0.05)；여저양조비교,각제량갈근소간예조세포활성화S기세포비례하강,Caspase-3활력승고,Kv 1.5단백급mRNA표체상조,차이균유통계학의의(P＜0.05).결론 갈근소가억제저양성PASMCs증식,촉진기조망,기작용궤제가능통과상조Kv 1.5표체억제PASMCs적증식.
Objective To study the effects of puerarin on proliferation,apoptosis and Kv1.5 gene expression of rat pulmonary artery smooth muscle cells (PASMCs) induced by hypoxia.Methods The rat PASMCs were divided into 5 groups:control group,hypoxia group,hypoxia plus puerarin ( 1×10-5 mol/L) group,hypoxia plus puerarin ( 1 ×10-4 mol/L) group and hypoxia plus puerarin ( 1 ×10-3 mol/L) group,and cultured at 37 ℃ for 24 h.The proliferation of rat PASMCs was detected by CCK-8 assay and flow cytometry,the activity of caspase-3 was measured with spectrophotometric method,Kv1.5 protein was detected by western blot,Kv1.5 mRNA was detected by real-time PCR.Results The cell viability and proportion of synthesis phase in control group were 0.940±0.045 and 9.67％± 1.28％,which were significantly lower than those ( 1.296±0.034 and 18.19％± 1.19％ )in hypoxia group (P＜0.05).The Caspase-3 activity,Kv 1.5 protein and Kv 1.5 mRNA in control group were 0.1073±0.0113,0.886±0.038 and 0.0377±0.0031,which were significantly higher than those (0.0664±0.0049,0.602±0.064 and 0.0108±0.0014 ) in hypoxia group (P＜0.05).As compared with hypoxia group,the cell viability and proportion of synthesis phase in 3 hypoxia plus puerarin groups significantly decreased,and the Caspase-3 activity,Kv 1.5 protein and Kv 1.5 mRNA in 3 hypoxia plus puerarin groups significantly enhanced (P＜0.05).Conclusion Puerarin could decrease the proliferation and increase the apoptosis induced by hypoxia in rat PASMCs,and the up-regulated expression of Kv1.5 gene may be the mechanism of puerarin effects.
