加味芍药甘草汤对人子宫腺肌病病灶细胞超微结构及细胞凋亡的影响  

Effect of Modified Shaoyao Gancao Decoction on Ultrastructure and Apoptosis of Human Adenomyosis Cells in Vitro

作　　者：凌静[1] 关永格[2] 李坤寅[1] 王帅[1] 

机构地区：[1]广州中医药大学,广东广州510006 [2]广州中医药大学第一附属医院,广东广州510405

出　　处：《广州中医药大学学报》2014年第4期601-606,680,681,682,共9页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(编号:81173296);广东省自然科学基金项目(编号:S2011010001939)

摘　　要：【目的】观察加味芍药甘草汤对人子宫腺肌病病灶细胞超微结构及凋亡率的影响。【方法】采用胶原酶消化法培养人子宫腺肌病病灶细胞,采用免疫细胞化学方法鉴定子宫腺肌病病灶细胞,将加味芍药甘草汤分为高、中、低浓度组(浓度分别为40、20、10 mg/mL),另设空白对照组及米非司酮组(浓度为1×10-5mol/L),各组药物作用于子宫腺肌病病灶细胞24、48 h后,采用透射电镜观察细胞超微结构,流式细胞术检测细胞凋亡率。【结果】电镜下加味芍药甘草汤及米非司酮干预后的细胞出现细胞核形态改变,胞浆中溶酶体增多,线粒体及内质网扩张等细胞凋亡早期特点。各组药物干预24、48 h后细胞凋亡率显著升高,与空白对照组比较差异均有统计学意义(P<0.05);米非司酮组、加味芍药甘草汤高浓度组细胞凋亡率与加味芍药甘草汤中、低浓度组比较差异均有统计学意义(P<0.05);加味芍药甘草汤各浓度组干预后24 h与48 h比较,差异均有统计学意义(P<0.05),具有一定的量效与时效关系。【结论】加味芍药甘草汤治疗子宫腺肌病的作用机制可能与改变人子宫腺肌病病灶细胞超微结构以提高细胞凋亡率有关。Objective To observe the effect of modified Shaoyao Gancao Decoction （MSGD） on the uhrastructure and apoptosis of hmnan adenomyosis cells in vitro. Methods Focal human adenomyosis tissues were digested with collagenase I for primary culture, and then the isolated adenomyosis cells were identified by immunocytochemistry. Adenomyosis cells were seperately treated with MSGD at the concentration of 40, 20, 10 mg/mL （MSGD groups） , DMEM （blank control group）, or mifepristone at the concentration of 1×10^-5 mol/L （ positive control group） for 24 and 48 hours. The uhrastructure of adenomyosis cells after intervention by MSGD and mifepristone was observed by transmission electron microscope, and the apoptotic rate was evaluated by flow cytometry （FCM） . Results Under transmission electron microscope, adenomyosis cells had condensed chromatin, shrinked nucleus, increased lysosome count in cytoplasm, dilated mitochondria and endoplasmic reticulum in MSGD groups and mifepristone group. After treatment with mifepristonc and MSGD for 24 and 48 hours, the apoptotic rate was significantly increased （P〈0.05 compared with blank control group）. The apoptotie rate in mifepristone group and high-concentration MSGD group were significantly higher than that in medium- and low-concentration MSGD groups after culture for 24 and 48 hours （P〈0.05）. MSGD groups after culture for 48 hours had higher apoptotic rate than that after 24 hours （P〈0.05）, showing certain dose- and time-dependent effect. Conclusion The therapeutic mechanism of MSGD is probably related with the alteration of ultrastructure of human adenomyosis cells and with the increase of their apoptotic rate.

关 键 词：子宫腺肌病 中药疗法 加味芍药甘草汤 药理学 原代细胞培养 人子宫腺肌病病灶细胞 超微结构 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]