瞬时感受器电位离子通道香草素受体4在睾丸缺血再灌注损伤中对GC-1细胞增殖及凋亡的作用及其机制

目的:探讨瞬时感受器电位离子通道香草素受体4(TRPV4)在睾丸缺血再灌注损伤(IRI)中对GC-1细胞增殖和凋亡的作用及其机制。方法:建立睾丸GC-1细胞缺氧复氧模型，采用蛋白质印迹法(Western blot)检测不同复氧损伤时间点TRPV4的表达变化；分别采用噻唑蓝(MTT)实验、流式细胞术检测转染TRPV4对G...

Effect of transient receptor potential vanilloid 4 on proliferation and apoptosis of GC-1 cells in testicular ischemia-reperfusion injury and mechanism

Objective To investigate the effect of transient receptor potential vanilloid 4(TRPV4)on the proliferation and apoptosis of GC-1 cells in testicular ischemia-reperfusion injury(IRI)and the underlying mechanism.Methods The hypoxia reoxygenation model of testicular GC-1 cells was established.Western blotting was used to detect the protein expression of TRPV4 at different reoxygenation injury time points.Methyl thiazolyl tetrazolium(MTT)assay and flow cytometry were used to measure the effect of TRPV4 transfection on the proliferation and apoptosis of GC-1 cells.Western blotting was used to examine the protein expression level of Caspase-3 and cytoc hrome C(Cyt-C)after TRPV4 transfection.T test was used for comparison between groups,and one-way analysis of variance(ANOVA)was used for comparison among multiple groups.Results The protein expression level of TRPV4 in the normoxia group and hypoxia reoxygenation group(0,6,12,24,48 and 72 h)was(0.19±0.02,0.35±0.03,0.42±0.04,0.46±0.04,0.62±0.05,0.54±0.05,0.45±0.04),respectively.The expression of TRPV4 in the hypoxia reoxygenation group was significantly higher than that in the control group,and reached the peak at hypoxia 3 h/reoxygenation 24 h(F=6.898,P<0.05).The level of cell proliferation in the hypoxia reoxygenation group was significantly lower than that in the control group(52.32±4.58 vs.100.00±7.63,t=-9.280,P<0.05),and the level of apoptosis in the hypoxia reoxygenation group was higher than that in the control group(15.60±1.72 vs.4.08±0.87,t=10.352,P<0.05).The level of cell proliferation in TRPV4 overexpression group was lower than that in control group(23.65±3.98 vs.51.35±4.67,t=-7.820,P<0.05),and the level of apoptosis in TRPV4 overexpression group was higher than that in control group(26.93±2.15 vs.14.62±1.68,t=7.814,P<0.05).The level of cell proliferation in TRPV4 silencing group was higher than that in control group(72.49±6.21 vs.53.18±5.14,t=4.150,P<0.05),and the level of apoptosis was lower than that in control group(9.71±1.25 vs.15.07±1.64,t=-4.502,P<0.05).The expression levels of Caspase-3 and Cyt-C in hypoxia reoxygenation group were higher than those in control group(0.70±0.06 vs.0.20±0.02,t=13.693,P<0.05;0.74±0.07 vs.0.26±0.03,t=10.917,P<0.05).The expression levels of Caspase-3 and Cyt-C in TRPV4 overexpression group were higher than those in control group(1.25±0.11 vs.0.69±0.07,t=7.439,P<0.05;1.38±0.14 vs.0.72±0.07,t=7.303,P<0.05).The expression levels of Caspase-3 and Cyt-C in TRPV4 group were lower than those in control group(0.46±0.05 vs.0.68±0.06,t=-4.879,P<0.05;0.45±0.05 vs.0.72±0.06,t=-5.988,P<0.05).Conclusion TRPV4 is highly expressed in GC-1 cells,which may change proliferation and apoptosis of GC-1 cells,thus affecting the development of testicular IRI.