线粒体途径在吡咯喹啉醌抑制氧化应激诱导施万细胞凋亡中的作用

Pyrroloquinoline quinone inhibited oxidative stress induced-apoptosis of Schwann cells via mitochondrial apoptotic signaling pathway in vitro

目的探讨线粒体凋亡途径在吡咯喹啉醌（pyrrUloquinolinequinone，PQQ）抑制过氧化氢（H2O2）诱导施万细胞（schwannceils，SCs）凋亡中的作用及机制。方法体外原代培养、纯化及S-100免疫荧光染色鉴定大鼠SCs。将培养的SCs分为空白对照组、H2O2诱导组（100μmol／L H2O2）、H202加PQQ处理组（10、50、100nmol／LPQQ）。CCK-8法检测SCs的增殖情况；AnnexinV—FITC／PI流式细胞术检测细胞凋亡；JC-1流式细胞术检测线粒体膜电位；Westernblot检测SCs内细胞色素c（cytochromeC，CytC）、Bax及Caspase-9蛋白表达。结果本实验培养细胞经S-100免疫荧光染色鉴定阳性率达95％以上，SCs经H2O2处理后细胞增殖明显受到抑制，凋亡增加，线粒体膜电位降低，CytC、Bax、Caspase-9表达增加；加入PQQ处理后细胞增殖活性增加，凋亡降低，线粒体膜电位部分恢复，CytC、Bax、Caspase-9表达降低。结论PQQ通过抑制线粒体凋亡途径，降低H2O2诱导的SCs凋亡。

Objective To investigate the mechanisms of Pyrroloquinoline quinone （PQQ） against oxidative stress induced apoptosis in Scbwann ceils （SCs）. Methods SCs were cultured in vitro, identified by S-100 immunofluorence staining. SCs were divided into control group, H2O2 induced group, H2O2 ＋ PQQ treated group. CCK-8 assay was used to detect cell proliferation. Apoptosis was detected by flow cytometry with Annecin V-FITC/PI staining, mitoehondrial transmembrane potential was detected by flow cytometry with JC-1 labeled staining, eytochrome C （CytC） , Bax and Caspase-9 protein levels was detected by Western blot analysis. Results In this study, the S-100 positive ceils were more than 95% , cell proliferation was decreased in H2O2 induced SCs, apoptotic rate was increased, mitochondrial transmembrane potential was decreased, CytC, Bax and Caspase-9 protein levels were increased. After PQQ added, cell proliferation was increased, apoptotic rate decreased, mitochondrial transmembrane potential increased, CytC, Bax and Caspase-9 protein levels decreased. Conclusions PQQ protects SCs from oxidative induced apoptosis by inhibiting mitochondrial signaling pathway.