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目的:观察雷公藤红素诱导人肥大细胞系(HMC-1)细胞凋亡过程中细胞形态的变化,并探讨其意义。方法:在HMC-1细胞培养体系中加入雷公藤红素,用光镜、电镜观察细胞的形态变化,用TUNEL和流式细胞术(FCM)检测细胞凋亡。结果:细胞在雷公藤红素作用下呈葵样改变:细胞母体居中,周边围以形成或正在形成的凋亡小体。葵样细胞(sunflower-likecel,SLC)对锥虫蓝拒染,可伴有核染色质浓缩和着边,药物作用20min即可形成,持续数小时后完全裂解为凋亡小体,或凋亡小体游离,残留细胞母体,最后崩溃;SLC的形成具有时间和药物浓度依赖性;在8h内与TUNEL标记和碘化丙啶染色/FCM检测的凋亡结果有显著相关性;SLC清晰易辨,个体内和个体间计数重复性良好。结论:SLC是凋亡细胞的特征性形态改变,可作为检测HMC-1细胞凋亡的定性和定量指标。
OBJECTIVE: To observe the changes of cell morphology during tripterine-induced human mast cell line (HMC-1) apoptosis and to explore its significance. Methods: Tripterine was added into HMC-1 cell culture system. The morphological changes of cells were observed with light microscope and electron microscope. Cell apoptosis was detected by TUNEL and flow cytometry (FCM). Results: The cells under the action of triproline showed the changes of sunflower-like cells: the cells were surrounded by the mother cells and the surrounding bodies formed or were forming apoptotic bodies. Asparagus blue was rejected by sunflower-like cells (SLC), which could be condensed and stained with chromatin. The drug could be formed after 20 minutes of treatment and completely lysed to apoptotic bodies or apoptosis after several hours Small body free, residual cell matrix, and finally collapse; the formation of SLC with time and drug concentration-dependent; within 8h and TUNEL markers and propidium iodide staining / FCM detection of apoptosis was significantly related; SLC clear , Individual and individual count repeatability is good. Conclusion: SLC is a characteristic morphological change of apoptotic cells, which can be used as a qualitative and quantitative indicator of apoptosis in HMC-1 cells.