School of Kinesiology, Lakehead University, Thunder Bay, Canada.
Cardiovasc Res. 2010 Jun 1;86(3):487-95. doi: 10.1093/cvr/cvp420. Epub 2010 Jan 5.
Cystathionine gamma-lyase (CSE)-derived H2S plays an important role in regulating cell growth. Lack of CSE expression results in development of hypertension. The current study compared proliferation of smooth muscle cells derived from CSE gene knockout mice (SMCs-KO) with that of wild-type mice (SMCs-WT).
Cell proliferation was assessed by bromodeoxyuridine incorporation. Gene expression was analysed by western blotting, real-time PCR, and microarray analysis. Enhanced cell proliferation was detected in SMCs-KO and in the media of the aorta from CSE KO mice. SMCs-KO underwent significantly more apoptosis than SMCs-WT when treated with exogenous H2S (100 microM). CSE KO mice showed much lower level of phosphorylated extracellular signal-regulated kinase (ERK1/2) in mesentery arteries compared with those of WT mice, and exogenous H2S induced more phosphorylation of ERK1/2 in SMCs-KO compared with that in SMCs-WT. Decreased p21(Cip/WAF-1) but increased cyclin D1 expression was observed in isolated SMCs and vascular tissues from CSE KO mice, and exogenous H2S caused more increase in p21(Cip/WAF-1) expression and more decrease in cyclin D1 expression in SMCs-KO than in SMCs-WT. The transcriptional expression of calcitonin receptor-like, intergrin beta 1, and heparin-binding epidermal growth factor-like growth factor was also significantly increased in the aorta of CSE KO mice.
SMCs-KO display an increased proliferation rate in vitro and in vivo, and these cells are more susceptible to apoptosis induced by exogenous H2S at physiologically relevant concentrations. These cellular effects of H2S are mediated by phosphorylation of ERK1/2 and altered expression of cyclin D1 and p21(Cip/WAF-1).
半胱氨酸γ-裂解酶（CSE）衍生的 H2S 在调节细胞生长中发挥重要作用。缺乏 CSE 表达会导致高血压的发展。本研究比较了 CSE 基因敲除小鼠（SMCs-KO）平滑肌细胞与野生型小鼠（SMCs-WT）的增殖情况。
通过溴脱氧尿苷掺入评估细胞增殖。通过 Western 印迹、实时 PCR 和微阵列分析分析基因表达。在 SMCs-KO 和 CSE KO 小鼠主动脉中检测到增强的细胞增殖。与 SMCs-WT 相比，用外源性 H2S（100 μM）处理时，SMCs-KO 经历了更多的细胞凋亡。与 WT 小鼠相比，CSE KO 小鼠肠系膜动脉中磷酸化细胞外信号调节激酶（ERK1/2）的水平低得多，而外源性 H2S 诱导 SMCs-KO 中 ERK1/2 的磷酸化程度高于 SMCs-WT。在分离的 SMC 和 CSE KO 小鼠的血管组织中观察到 p21（Cip/WAF-1）表达降低而 cyclin D1 表达增加，外源性 H2S 引起 SMCs-KO 中 p21（Cip/WAF-1）表达增加更多，而 cyclin D1 表达降低更多。在 CSE KO 小鼠的主动脉中，降钙素受体样、整合素β 1 和肝素结合表皮生长因子样生长因子的转录表达也显著增加。
SMCs-KO 在体外和体内显示出更高的增殖率，并且这些细胞在生理相关浓度的外源性 H2S 诱导下更容易发生凋亡。H2S 的这些细胞效应是通过 ERK1/2 的磷酸化以及 cyclin D1 和 p21（Cip/WAF-1）的表达改变来介导的。
