硫化氢对正常及自发性高血压大鼠主动脉平滑肌细胞PCNA和p-ERK蛋白表达的影响

目的研究硫化氢(hydrogen sulfide,H2S)对正常大鼠及自发性高血压大鼠(spontaneous hypertensive rat,SHR)主动脉平滑肌细胞(aortic smooth muscle cell,ASMC)增殖的影响。方法分别将正常大鼠及SHR主动脉平滑肌细胞分为实验组和对照组,实验组中分别加入浓度为(10-5~10-3)mol.L-1H2S供体的硫氢化钠(sodium hydrosulfide,NaHS)或浓度为(2~10)mmol.L-1胱硫醚-γ-裂解酶抑制剂的炔丙基甘氨酸(DL-propargy lglycine,PPG),对照组加入等体积的生理盐水,分别以蛋白质印迹法观察6、12、18和24h时间点的平滑肌细胞中细胞增殖核抗原(PCNA)和磷酸化的细胞外信号调节激酶(p-ERK)蛋白表达情况。结果NaHS对正常大鼠ASMC增殖及p-ERK蛋白表达无影响。PPG显著促进正常大鼠ASMC增殖及p-ERK蛋白表达。NaHS显著抑制SHR的ASMC增殖及p-ERK蛋白表达。PPG显著促进SHR的ASMC增殖及p-ERK蛋白表达。结论内源性H2S下调可促进正常大鼠及SHR的ASMC增殖,而外源性补充H2S可抑制SHR的ASMC异常增殖,ERK信号途径可能介导H2S抑制ASMC增殖的分子机制。

Effect of hydrogen sulfide on the expression of PCNA and p-ERK in the aortic smooth muscle cells of normal rats and spontaneous hypertensive rats

Aim To investigate the effect of hydrogen sulfide(H2S)on the proliferation of the aortic smooth muscle cells(ASMCs)of normal rats and spontaneous hypertensive rats(SHRs).Methods ASMCs of normal rats and SHR were cultured and divided into experimental group and control group,respectively.Sodium hydrosulfide(NaHS,an H2S donor)at the concentration of(10-5~10-3)mol·L-1 or DL-propargylglycine(PPG,an inhibitor of cystathionine-γ-lysase)at the concentration of(2~10)mmol·L-1 was added into the experimental group with normal saline of equal volume as control.After being cultured for 6,12,18 h and 24 h,expression of proliferative cell nuclear antigen(PCNA)and phosphorylated extracellular signal-regulated kinase(p-ERK)in different group ASMCs were detected by Western blot.Results NaHS had no effect on the proliferation of ASMCs of normal rats and the expression of p-ERK.PPG promoted the proliferation of ASMCs of normal rats and the expression of p-ERK obviously.NaHS inhibited the proliferation of ASMCs of SHRs and the expression of p-ERK obviously.PPG increased the proliferation of ASMCs of SHRs and the expression of p-ERK obviously.Conclusions Downregulation of endogenous H2S could promote the proliferation of ASMCs of normal rats and SHR,while supplement of exogenous H2S could inhibit the abnormal proliferation of ASMCs of SHR.ERK pathway might be involved in the mechanism for H2S inhibiting proliferation of ASMCs.