钾通道与肺动脉平滑肌细胞增殖和凋亡关系中钙通道所起作用的研究

Role of calcium channel in relationship between potassium channel and proliferation and apoptosis of pulmonary artery smooth muscle cells

目的:观察钾通道与肺动脉平滑肌细胞(PASMC)增殖和凋亡关系中钙通道所起的作用。方法:将PASMC分为对照、钾通道阻断剂四乙胺TEA(开放剂比那地尔Pin)、钾通道阻断剂(开放剂)+钙通道阻断剂硝苯地平Nif(开放剂BayK8644)组,采用非核素增殖试剂盒、3HTdR掺入测定细胞增殖。将PASMC分为对照、钾通道开放剂、钾通道开放剂+钙通道开放剂组,采用细胞凋亡检测试剂盒测定细胞凋亡。结果:与对照组相比,10～20mmol/L的钾通道阻断剂TEA显著增加PASMC细胞数(P<0.01),15～30mmol/LTEA显著增加DNA合成(P<0.05,P<0.01)。TEA+5μmol/LNif组中只有20mmol/LTEA使DNA合成增加(P<0.05),其余浓度组与对照组均无统计学差异。TEA+Nif组与TEA组相比,TEA浓度在10～25mmol/L时2组细胞数相差显著(P<0.01),15～30mmol/LTEA时2组DNA合成相差显著(P<0.05,P<0.01)。与对照组相比,25～500μmol/LPin显著减少PASMC细胞数和DNA合成(P<0.01)。Pin+1μmol/LBay组与Pin组相比,当Pin浓度为25～500μmol/L时2组PASMC细胞数相差显著(P<0.01),当Pin浓度在50～500μmol/L时DNA合成相差显著(P<0.01)。Pin+Bay组与对照组相比,50～500μmol/L的Pin仍能显著减少PASMC细胞数(P<0.01),25～500μmol/L的Pin显著减少DNA合成(P<0.01)。与对照组相比。

Objective:To observe the role of calcium channel in the relationship between potassium channel and proliferation and apoptosis of pulmonary artery smooth muscle cells (PASMC).Methods: To detect cell proliferation, PASMC were divided into control, potassium channel blocker TEA (opener Pin), and potassium channel blocker (opener)+calcium channel blocker Nif (opener Bay K8644) groups, where cell proliferation was observed with a non-isotope proliferation kit and 3H-TdR incorporation. To detect cell apoptosis, PASMC were divided into control, potassium channel opener, potassium channel opener+calcium channel opener groups, where cell apoptosis was observed with a cell apoptosis test kit. Results: TEA significantly increased the cell number at 10-20 mmol/L (P<0.01) and DNA synthesis at 15-30 mmol/L (P<0.05,P<0.01) compared with the control group. There were no statistical differences between the groups of different concentrations of TEA+5 μmol/L Nif and the control group, except that TEA at 20 mmol/L significantly increased DNA synthesis (P<0.05). There were significant differences between TEA+Nif and TEA group only in cell number at 10-25 mmol/L (P<0.01) and DNA synthesis at 15-30 mmol/L(P<0.05,P<0.01). Pin at 25-500 mmol/L significantly decreased the cell number and DNA synthesis of PASMC compared with the control group (P<0.01). There were significant differences between Pin+1 μmol/L Bay group and the Pin group in cell number(Pin at 25-500 mmol/L) and DNA synthesis (Pin at 50-500 mmol/L) (P<0.01). In Pin+Bay group, Pin at 50-500 μmol/L significantly decreased the cell number and DNA synthesis at 25-500 mmol/L compared with the control group(P<0.01). Pin at 50-500 μmol/L significantly increased the apoptosis rate of PASMC compared with the control group (P<0.05, P<0.01). There were no significant differences in apoptosis rate between Pin+1 μmol/L Bay group and Pin group. Conclusion: Calcium channel plays an important role in the relationship between potassium channel and proliferation of PASMC, though has no obvious effect in the relationship between potassium channel and apoptosis of PASMC.