氯化镉和氯化锌对V79细胞凋亡的影响

Study on Apoptosis Induced by CdCl_2 Alone or with ZnCl_2 in vitro

目的通过体外实验,以两种简便方法了解氯化镉(CdCl2)、氯化锌(ZnCl2)对中国仓鼠肺成纤维细胞(V79)细胞凋亡的影响.方法采用MTT法了解CdCl2对于V79细胞的毒性;在此基础上采用碱性单细胞凝胶电泳法和激光共聚焦技术结合AO/EB双染法,研究不同剂量CdCl2对V79细胞凋亡的影响,并采用生理浓度的ZnCl2与CdCl2同时作用后观察细胞凋亡情况.结果 MTT实验中,CdCl2对于V79细胞的毒性随染毒剂量增加而增强,呈线性关系;碱性单细胞凝胶电泳法和AO/EB双染法得到相似的结果,即1～32 μmol/L CdCl2可以引起V79细胞凋亡,凋亡率、凋亡指数分别为3.0%～49.6%及3.5%～53.8%,且随剂量增加而增大,呈剂量-效应关系;凋亡率、凋亡指数与CdCl2染毒浓度间可拟合成直线方程:y=0.931 5x-0.415 2.10 μmol/L ZnCl2可以拮抗CdCl2所致的凋亡效应.结论在本实验条件下,采用单细胞凝胶电泳和激光共聚焦技术均可以灵敏、快捷地检测CdCl2对于V79细胞的凋亡作用和锌对镉所致细胞凋亡的拮抗效应.

Objective To investigate apoptosis of Chinese hamster lung fibroblast cells （V79） exposed to CdCl2 alone or with ZnCl2 with two simple and novel assays. Methods MTT assay was used to detect the cytotoxicity of CdCl2 on V79 cell line. By SCGE assay and AO/EB bi-label assay plus Confocal Scanning Laser Microscope （CSLM）, the apoptotic effects induced by CdCl2 alone or with 2inc simultaneously were observed. Results Dosage of CdCl2 range in 5 - 40μmol/L could inhibit the cellular proliferation effectively after treatment for 24 h, the effect was in a dose-dependent pattern. In alkali SCGE assay and AO/EB bi-label assay, CdCl2 could induce apoptosis in V79 cells in a linear manner within the dosage range used, while 10μmol/L ZnCl2 could protect cells from the induction of CdCl2. The results of two assays match well. Conclusions SCGE and CSLM could be two good ways to detect the apoptotic effects induced by CdCl2 and the anti-apoptotic effects caused by ZnCl2.