摘要
目的:探讨钙敏感受体(CaSR)在缺氧诱导的大鼠肺动脉平滑肌细胞(PASMCs)增殖中的信号转导途径。方法:Ⅱ型胶原酶消化法提取、培养大鼠PASMCs。通过缺氧培养箱内(93%N2、2%O2、5%CO2)培养24 h的方法复制细胞缺氧模型。应用Western blotting分析不同处理情况下细胞周期素D1(cyclin D1)及磷酸化的蛋白激酶B(p-Akt)在PASMCs中的表达;采用流式细胞术检测不同处理因素对细胞增殖周期及增殖指数的影响,应用BrdU掺入法分析不同处理因素对细胞DNA合成的影响。结果:缺氧引起PASMCs的cyclin D1及p-Akt表达水平上调,增加BrdU掺入量、S期细胞数量和细胞增殖指数,GdCl3能够放大缺氧的作用,但上述效应可以被LY294002抑制。结论:CaSR通过PI3K/Akt信号通路参与缺氧诱导的大鼠PASMCs增殖。
AIM ： To explore the signal transduction pathways of calcium sensing receptor （CaSR） that medi ates hypoxia induced proliferation of rat pulmonary artery smooth muscle cells （PASMCs）. METHODS： The expression of cyclin DI and phosphorylated protein kinase B （p Akt） was analyzed by Western blotting. Cell proliferation was tested using a BrdU incorporation assay, and cell cycle analysis was carried out using a flow cytometric assay. RF^ULTS： Hy poxia significantly increased the expression of cyclin D1 and p Akt, the BrdU incorporation and the cell proliferation in dex. GdCl3, an agonist of CaSR, amplified the effect of hypoxia. LY294002,a PI3K inhibitor, decreased the up - regula tion of cyclin D1 expression and the BrdU incorporation, and also inhibited the increase in the cell proliferation index in duced by hypoxia and GdCI3 in PASMCs. CONCLUSION： The CaSR mediates hypoxia - induced proliferation of rat PASMCs through PI3K pathways.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2012年第9期1549-1553,共5页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.30871012
No.81070123)
黑龙江省教育厅资助项目(No.12511618)
关键词
受体
钙敏感
肺动脉平滑肌细胞
缺氧
信号转导
Receptors, calcium - sensing
Pulmonary arterial smooth muscle cells
Hypoxia
Signal transduc- tion
