MSCs上清可减轻雷公藤甲素对KGN细胞的损伤作用.pdfVIP

周 雪等 MSCs上清可减轻雷公藤甲素对KGN细胞的损伤作用 第 12期 · 1641 · doi：10．3969／j．issn．1000-484X．2014．12．012 MSCs上清可减轻雷公藤 甲素对 KGN细胞 的损伤作用① 周 雪 赵光锋 陈士雯 侯亚义 (南京大学医学院免疫与生殖生物学实验室，南京 210093) 中图分类号 11392 文献标志码 A 文章编号 1000-484X(2014)12-1641-06 [摘 要] 目的：探讨MSCs上清对雷公藤甲素损害卵巢颗粒细胞的缓解作用及机制。方法：用 CCK一8法分析了雷公藤 甲素对 KGN活力的影响；用混合酶消化法分离脐带来源的MSCs，并用流式细胞术对其表型进行了鉴定；收集 MSCs上清，用 CCK一8法及PI法检测 MSCs上清对雷公藤甲素损伤的KGN活力及周期的影响；用 Real—timePCR法对周期调节相关基 因 CDKN1A的表达进行检测。结果：雷公藤甲素能够通过抑制KGN活力及阻滞 KGN在 s期抑制其生长；MSCs上清不影响正常 KGN的增殖和周期，但可提高雷公藤甲素损伤的KGN细胞的活力，缓解雷公藤甲素引起的S期阻滞 ，抑制雷公藤甲素对 KGN 的CDKN1A表达的异常上调。结论：MSCs上清可能减轻雷公藤甲素对KGN细胞的损伤作用。 【关键词] 间充质干细胞；卵巢颗粒细胞 ；雷公藤甲素；CDKN1A M esenchymalstem cel1．conditionedmedium could amelioratedtriptolideinduced damageinKGN cells ZHOU Xue．ZHAO Guang-Feng，CHEN Shi—Wen，HOU Ya—Yi．Immunology andReproductiveBiologyLaboratory， MedicalSchoolofNanjingUniversity，Nanfing210093，China [Abstract] Objective：ToexploretheameliorativeeffectandmachanismofMSCsconditionedmediumontheovariangranulosa cellsdamageinducedbytriptolide．M ethods：CellCountingKit8 assaywasused to examinethecellvitality ofKGNswith the treatmentoftriptolide．Themixedenzymedigestionmethodwereusedfortheisolationofhumanumbilicalcordmesenchymalstem cells (MSCs)，andflowcytometrywasusedforthesubsequentimmunotypeidentification．MSCsconditionedmediumwascollected，andCell CountingKit8assayandPIstainingwasusedtoanalysetheeffectofMSCsconditionedmedium onthecellvitalityandcellcycledistli— bution of triptolide-damaged KGN．Real-time PCR method was used to examine the expression of cell cycle related gene CDKN1A．Results：TriptolidecaninhibitKGN cellgrowthwiththeinhibitionofcellvitalityandcellcycleofKGN．MSCsconditioned medium didnotinfluencetheproliferationand cellcycleofnormalKGN，butimprovedthetriptolide-inducedvitalityinhibitionand extentofS-phasearrest，and inhibittheabnormalup—regulation ofCDKN1A in KGN．Conclusion：MSCsconditioned medium amelioratedtheKGN celldamageinducedb