理气活血中药对子宫腺肌细胞来源外泌体调控的研究  

Study on the regulation of exosome derived from adenomyocytes by Chinese herbs for regulating qi and activating blood circulation

作　　者：姜心禅 陈晓波 JIANG Xin-chan;CHEN Xiao-bo(College of Traditional Chinese Medicine,Guangdong Pharmaceutical University,Guangzhou 510000,China;The First Affiliated Hospital of Guangdong Pharmaceutical University,Guangzhou 510080,China)

机构地区：[1]广东药科大学中医学院,广州510000 [2]广东药科大学附属第一医院,广州510080

出　　处：《中华中医药杂志》2021年第8期5015-5018,共4页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：广东省中医药局面上科研项目(No.20191198);广东省医学科学技术研究基金项目(No.A2019368);广东省基础与应用基础研究基金项目(No.2019A1515111175)。

摘　　要：目的:研究理气活血中药治疗子宫腺肌病(AM)的机制。方法:将体外培养的子宫腺肌细胞经免疫组化法鉴定后随机分为4组:中药组、西药组、空白组和无外泌体组。经透射电镜观察外泌体(ex o)形态,Western blot检测exo膜表面蛋白CD63、CD9和calnexin,电镜观察各组细胞的超微结构,流式细胞术检测细胞凋亡的情况,RT-qPCR技术检测miR-21的表达。结果:AM细胞来源的exo为近圆形,直径约30~90nm,表面膜蛋白CD63、CD9为阳性、calnexin为阴性;电镜下可见中药组和西药组细胞有细胞凋亡的典型表现,空白组和无外泌体组无凋亡改变;与无外泌体组比较,空白组细胞凋亡率显著降低(P<0.05),m i R-21表达量显著升高(P<0.05);与空白组比较,两给药组细胞凋亡率显著升高(P<0.05),且细胞miR-21表达量均显著降低(P<0.05),但中药组与西药组之间差异无统计学意义。结论:理气活血中药能抑制exo对细胞增殖的促进作用,同时加快细胞凋亡,其作用机制可能与调控细胞中miR-21的表达相关。Objective:To study the mechanism of regulating qi and activating blood circulation in the treatment of adenomyosis(AM).Methods:The cultured adenomyocytes were randomly divided into four groups:traditional Chinese medicine group,western medicine group,blank group and no exosome group.The morphology of exosomes(exo)was observed by transmission electron microscopy,CD63,CD9 and calnexin were detected by Western blot,the ultrastructure of cells in each group was observed by electron microscopy,apoptosis was detected by flow cytometry,and miR-21 was detected by RT-qPCR.Results:The diameter of exo from am cells was about 30~90nm,surface membrane protein CD63 and CD9 were positive while calnexin was negative.Under electron microscope,there were typical apoptosis characteristics in the traditional Chinese medicine group and western medicine group,but no apoptosis in the blank group and the no exosome group.Compared with the no exosome group,the apoptosis rate of the cells in the blank group was lower(P<0.05),the expression of miR-21 in the blank group was higher(P<0.05).Compared with the blank group,the apoptosis rate in the traditonal Chinese medicine group and western group were increased(P<0.05),the expression of miR-21 in traditional Chinese medicine group and western medicine group were significantly decreased(P<0.05),but there was no significant difference between them.Conclusion:Chinese herbs for regulating qi and activating blood circulation can inhibit the promoting effect of exo on cell proliferation and accelerate cell apoptosis.The mechanism may be related to the regulation of miR-21 expression in cells.

关 键 词：理气活血法 外泌体 子宫腺肌病 机制 细胞增殖 细胞凋亡 MIR-21 

分 类 号：R285.5[医药卫生—中药学]