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目的探讨氯化钆对大鼠腹腔巨噬细胞内质网应激和凋亡的作用。方法分离和培养大鼠腹腔巨噬细胞(PMΦ),分别经氯化钆(Gd)和衣霉素(Tunicamycin)处理后,采用细胞免疫组化S-P法检测ARMET的表达,免疫荧光双标染色检测ARMET和CHOP的表达,PI染色观察对细胞凋亡的影响。结果氯化钆处理组细胞数目减少,胞体变小变圆;ARMET在氯化钆组(5×10-6mol/L)的阳性细胞百分数为(42.1±6.6)%,在正常对照组的阳性率为(6.1±1.6)%,差异有统计学意义(P<0.05);而其在阳性对照Tunicamycin组的阳性率为(43.4±5.9)%,两者差异无统计学意义(P>0.05);免疫荧光双标染色提示氯化钆组ARMET和CHOP均高表达;PI染色可见氯化钆组细胞被深染、胞核断裂等凋亡特征。结论氯化钆可诱导大鼠腹腔巨噬细胞内质网应激和凋亡,其作用机制可能与胞内钙超载引起的非折叠蛋白反应有关。
Objective To investigate the effect of gadolinium chloride on endoplasmic reticulum stress and apoptosis in rat peritoneal macrophages. Methods Peritoneal macrophages (PMΦ) were isolated and cultured. After treatment with gadolinium chloride (Gd) and tunicamycin, the expression of ARMET was detected by immunohistochemical SP method. ARMET and CHOP expression, PI staining observed the impact of apoptosis. Results The number of cells in gadolinium chloride group decreased and the cell body became smaller and round. The percentage of positive cells of ARMET in gadolinium chloride group (5 × 10-6 mol / L) was (42.1 ± 6.6)%, and in positive control group (6.1 ± 1.6)%, respectively. The positive rate was (43.4 ± 5.9)% in the group of positive control Tunicamycin, but the difference was not statistically significant (P> 0.05). Immunofluorescence double-stained staining showed high expression of ARMET and CHOP in gadolinium chloride group; apoptotic characteristics such as deep staining and nucleus rupture were observed in PI group. Conclusion Gadolinium chloride can induce endoplasmic reticulum stress and apoptosis in rat peritoneal macrophages, and its mechanism may be related to the non-folded protein reaction caused by intracellular calcium overload.