摘要
目的探讨血管平滑肌细胞增殖过程中蛋白激酶G与钙调磷酸酶信号通路之间是否存在交互调节作用。方法植块法原代培养Wistar大鼠血管平滑肌细胞。Western blot测定钙调磷酸酶和蛋白激酶G Iα蛋白的表达。定磷法测定钙调磷酸酶活性,MTT法测定血管平滑肌细胞的增殖。结果苯肾上腺素刺激诱发的平滑肌细胞钙调磷酸酶的表达和活性可被一氧化氮供体S-亚硝基-N-乙酰青霉胺和Sp-8-pCPT-cGMP抑制,但可被Rp-8-pCPT-cGMP增强。应用环孢素A干预增殖细胞后蛋白激酶G Iα蛋白表达量较对照组明显提高(P<0.01),而加入苯肾上腺素刺激后蛋白激酶G Iα蛋白的表达较对照组也有明显提高(P<0.05),但较5 mg/L环孢素A干预组减少了36.7%,两组之间差异有统计学意义(P<0.01)。经0.5 mg/L环孢素A预处理后,苯肾上腺素刺激诱发的平滑肌细胞吸光度降低36.67%,S-亚硝基-N-乙酰青霉胺、Sp-8-pCPT-cGMP或Rp-8-pCPT-cGMP并不能进一步抑制或升高已升高的吸光度。结论蛋白激酶G介导钙调磷酸酶信号抑制血管平滑肌细胞增殖,并且蛋白激酶G与钙调磷酸酶能够交互调节影响血管平滑肌细胞增殖进程。
Aim To research the cross regulation of cGMP-dependent protein kinase （PKG） and calcineurin （CAN） in vascular smooth muscle cells（SMC） proliferation. Methods Primary vascular SMC from rat aorta were used as the experimental model. Expression of CAN and PKG was assayed using immtmoblotting. Cell growth was determined by MTT assay. Results The results showed that phenylephrine （ PE）-induced expression and activity of CaN protein was reduced by S-nitroso- N-acetylpenicillamine （SNAP） and Sp-8-pCPT-cGMP, but increased by Rp-8-pCPT-cGMP. The OD ratio of PKG Iα mRNA expression in 0.5 mg/L cyclosporin A （CsA） group resembled control group while in 5 mg/L CsA group was significantly higher than control group （ P 〈 0.01 ）. Although the result of 5 mg/L CsA ＋ 10 μmol/L PE group was lower than 5 mg/L CsA group （the expression of mRNA decreased 32.2% ; the production of PKG Iα protein decreased 36.7%, P 〈 0.01 ）, but still higher than control group obviously （ P 〈 0.05 ）. In SMC pretreated with CsA, absorbance of cells stimulated by PE decreased by 36.67%, but it could not be further altered by the additional treatment of SNAP, Sp-8-pCPT-cGMP and Rp-8-pCPT-cGMP. Conclusion PKG and CaN can cross-regulate in vascular smooth mucle cells proliferation.
出处
《中国动脉硬化杂志》
CAS
CSCD
2006年第8期645-648,共4页
Chinese Journal of Arteriosclerosis
基金
国家重点基础研究发展项目(G2000056905)资助
