甘氨胆酸对小鼠体外脾淋巴细胞凋亡的影响

EFFECTS OF Glycocholic ACID ON SPLEEN LYMPHOCYTES APOPTOSIS IN VITRO IN MICE

本研究旨在探索不同浓度的甘氨胆酸(Glycocholic acid,GCA)对小鼠体外脾淋巴细胞凋亡的影响。在0.02μg/ml,0.035μg/ml,0.07μg/ml,0.21μg/ml,1.05μg/mL不同浓度的GCA作用下,应用倒置显微镜观察了体外脾淋巴细胞的核形态,采用琼脂糖凝胶电泳法检测了脾淋巴细胞的DNA片段,采用流式细胞仪测定了脾淋巴细胞凋亡早期磷脂酰丝氨酸外翻百分率;采用分光光度法测定了凋亡细胞氧化还原反应活性——GSH的含量;通过ELISA法检测了Caspase-3和Caspase-9的活性。结果:GCA能够使脾淋巴细胞细胞产生核固缩和凋亡小体;显著引起脾淋巴细胞DNA片段化(P<0.05),极显著的诱导脾淋巴细胞磷脂酰丝氨酸外翻(P<0.01);显著降低GSH的含量(P<0.05);显著提高Caspase-9和Caspase-3的活性(P<0.05)。结论:GCA具有显著的提高小鼠体外脾淋巴细胞凋亡的作用(P<0.05)。

The aim of the study is to explore effects of manmade extracted GCA of different concentration on spleen lymphocytes apoptosis in vitro in mice. After adding GCA of different concentration to spleen lymphocytes, the morphologic features of spleen lymphocytcs in vitro were observed under inverted microscope, agarose gel eiectrophoresis was used for examining DNA fragment of spleen lymphocytes, flow cytometry was used for examining the percentage of apoptosis on spleen lymphocytes which Serine had evaginated, the method of speetrophotometer was used for examining the activity of redox reaction after apply GCA - content of GSH, and ELISA was used for examining activity of Caspase - 9 and Caspase - 3. The results show that spleen lymphocytes produce karyopyknosis and apoptotic bodies after adding GCA of different concentration to spleen lymphocytes, DNA fragmentation was caused remarkably, and induce lymphcell in spleen Serine had evaglnated, apoptosis rate on spleen lymphocytes and activity of Caspase -9 and Caspase -3 were improved significandy, and also content of GSH was reduced obviously. In conclusion, GCA has remarkable improvement influence on apoptosis of spleen lymphocytes in vitro in mice.