CAJ | 학술논문

目的体外以缺氧无血清条件模拟心肌梗死后的心脏缺血微环境,研究洛伐他汀能否抑制缺氧无血清引起的骨髓间充质干细胞(MSC)凋亡并探讨其机制.方法以Hocchst33342染色荧光显微镜观察法及Annexin V/PI流式细胞术检测洛伐他汀的抗凋亡作用,并进一步采用Westernblot方法检测洛伐他汀对线粒体凋亡途径的抑制作用以及对磷脂酰肌醇3激酶(PI3K)/丝氨酸苏氨酸激酶(Akt)途径和丝裂原活化的蛋白激酶(MAPK)的激酶(MEK)/细胞内信号调节蛋白激酶(ERK1/2)途径的激活作用.结果 0.01～1 μmol/L浓度范围的洛伐他汀能够有效地抑制缺氧无血清引起的MSC凋亡.洛伐他汀抑制线粒体凋亡途径,洛伐他汀抑制细胞色素C释放,降低天冬氨酸特异性半胱氨酸蛋白酶-3(caspase-3)活化,从而保护线粒体功能.洛伐他汀的抗凋亡效应以及其抑制细胞色素C释放的作用均可被PI3K抑制剂LY294002和MEK抑制剂U0126阻断.洛伐他汀能够激活PI3K/Akt和MEK/ERK1/2两条细胞存活信号通路,分别导致Akt和GSK-3β及ERK1/2磷酸化.结论洛伐他汀能够抑制线粒体凋亡途径,并激活PI3K/Akt和MEK/ERK1/2细胞存活通路,最终发挥抗缺氧无血清引起的MSC凋亡.该研究为提高移植干细胞的存活率提供了一种可能有效的干预措施.
목적 체외이결양무혈청조건모의심기경사후적심장결혈미배경,연구락벌타정능부억제결양무혈청인기적골수간충질간세포(MSC)조망병탐토기궤제.방법 이Hocchst33342염색형광현미경관찰법급Annexin V/PI류식세포술검측락벌타정적항조망작용,병진일보채용Westernblot방법 검측락벌타정대선립체조망도경적억제작용이급대린지선기순3격매(PI3K)/사안산소안산격매(Akt)도경화사렬원활화적단백격매(MAPK)적격매(MEK)/세포내신호조절단백격매(ERK1/2)도경적격활작용.결과 0.01～1 μmol/L농도범위적락벌타정능구유효지억제결양무혈청인기적MSC조망.락벌타정억제선립체조망도경,락벌타정억제세포색소C석방,강저천동안산특이성반광안산단백매-3(caspase-3)활화,종이보호선립체공능.락벌타정적항조망효응이급기억제세포색소C석방적작용균가피PI3K억제제LY294002화MEK억제제U0126조단.락벌타정능구격활PI3K/Akt화MEK/ERK1/2량조세포존활신호통로,분별도치Akt화GSK-3β급ERK1/2린산화.결론 락벌타정능구억제선립체조망도경,병격활PI3K/Akt화MEK/ERK1/2세포존활통로,최종발휘항결양무혈청인기적MSC조망.해연구위제고이식간세포적존활솔제공료일충가능유효적간예조시.
Objective To investigated the effect of lovastatin on hypoxia and serum deprivation (Hypoxia/SD) induced rat MSCs apoptosis in vitro and associated signaling pathway changes. Methods MSCs were isolated from Sprague-Dawley rats. The anti-apoptotic effects of lovastatin were detected using Hoechst33342 and annexin V-FITC/PI binding assay by Flow cytometric analysis. The phosphorylation of Akt and ERK1/2, the cytochrome C and the cleaved caspase-3 were detected by Western blot. Results Lovastatin (0. 01 - 1 μmol/L)significantly reduced Hypoxia/SD-induced MSCs apoptosis and increased Akt phosphorylation, reduced caspase-3 activation and cytochrome c release from mitochondria to cytosol in a time dependent manner. These effects could be significantly blocked by both PI3K inhibitor, LY294002 and ERK1/2 inhibitor, U0126. Conclusions Our results showed that lovastatin protects MSCs from Hypoxia/ SD-induced apoptosis via activating PI3K/Akt and ERK1/2 signaling pathways suggesting a potential role of statins as an adjunct therapeutic agent during transplanting MSCs into damaged heart after myocardial infarction.