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目的观察RH株弓形虫感染小鼠血清在体外对小鼠B16黑色素瘤细胞增殖以及凋亡的影响。方法B16细胞在含5%、10%和20%RH株弓形虫感染小鼠血清RPM1640中培养24和48 h,四甲基氮噻唑蓝(MTT)法检测B16细胞增殖抑制率;在10%、20%感染小鼠血清中培养24 h,Annexin V/PI染色,流式细胞检测细胞凋亡,HE染色观察细胞形态改变。结果正常血清能促进B16细胞增殖。5%、10%和20%弓形虫感染小鼠血清与B16细胞共孵育24和48 h,细胞生长抑制率分别为(9.06±0.36)%(、14.77±0.94)%(、23.74±0.5)%和(14.82±0.77)%(、24.56±1.04)%、(39.77±2.82)%,与对照组比较差异有统计学意义(P<0.05);10%、20%弓形虫感染鼠血清组B16细胞24 h凋亡率分别为(26.01±3.27)%和(44.55±2.87)%,显著高于对照组凋亡率(5.01±2.62)(P<0.05)。弓形虫感染鼠血清作用的B16细胞生长密度明显降低,细胞核固缩、浓染。结论RH株弓形虫感染血清能够抑制B16细胞增殖并促进其凋亡。
Objective To observe the effect of serum of mice infected with Toxoplasma gondii on proliferation and apoptosis of mouse B16 melanoma cells in vitro. Methods B16 cells were cultured in RPM1640 containing 5%, 10% and 20% RH Toxoplasma gondii strains for 24 and 48 hours. The inhibitory rate of B16 cells proliferation was detected by MTT assay. Twenty (20%) infected mice were cultured in serum for 24 h. The cells were stained with Annexin V / PI. The apoptosis of the cells was detected by flow cytometry. The morphology of the cells was observed by HE staining. Results Normal serum can promote the proliferation of B16 cells. The cell growth inhibition rate of the mice infected with Toxoplasma gondii with 5%, 10%, and 20% of Toxoplasma gondii were (9.06 ± 0.36)% (14.77 ± 0.94)% (23.74 ± 0.5)% And (14.82 ± 0.77)% (24.56 ± 1.04)% and (39.77 ± 2.82)%, respectively, which were significantly different from the control group (P <0.05) The apoptotic rates at 24 h were (26.01 ± 3.27)% and (44.55 ± 2.87)%, respectively, which were significantly higher than those in control group (5.01 ± 2.62) (P <0.05). The growth of B16 cells infected with Toxoplasma gondii sera was significantly reduced, and the nuclei were pyknotic and dense. Conclusion Toxoplasma gondii infection with serum can inhibit the proliferation of B16 cells and promote its apoptosis.