硼对大鼠脾脏淋巴细胞的毒性作用及增殖和凋亡的影响

Effects of Boron on the Cytotoxicity,Proliferation and Apoptosis of Spleen Lymphocytes in Rats

目的:研究不同浓度硼对大鼠脾脏淋巴细胞毒性作用,以及对其增殖和凋亡的影响。方法:选用2月龄清洁级Sprague Dawley(SD)雄性大鼠,无菌分离脾脏淋巴细胞,将其分为对照组和试验Ⅰ~组,分别在淋巴细胞中添加0、0.1、0.2、0.4、0.8、1、2、4、8、10、20、40、80、100mmol/L的硼酸,处理24h后,MTT和Cell-Counting Kit-8法检测硼对淋巴细胞的毒性作用,流式细胞仪检测淋巴细胞增殖和凋亡情况。结果:细胞毒性试验表明,与对照组相比,试验Ⅲ组淋巴细胞抑制率显著降低(P<0.05),增殖率显著升高(P<0.05),而试验Ⅹ~I组淋巴细胞抑制率极显著的升高(P<0.01),增殖率极显著的降低(P<0.01);流式细胞仪检测表明,试验Ⅲ组淋巴细胞增殖率较对照组极显著升高(P<0.01),凋亡率显著降低(P<0.05),而试验Ⅺ和Ⅻ组凋亡率均极显著升高(P<0.01),增殖率均极显著降低(P<0.01)。结论:添加0.4mmol/L的硼酸对淋巴细胞没有毒性作用,可以显著刺激淋巴细胞的增殖,抑制其凋亡,当添加量达到20mmol/L时,则对淋巴细胞产生明显的毒性作用,提高淋巴细胞的凋亡率。

Objective:To study the effects of different concentrations of boron on the toxicity,proliferation and apoptosis of spleen lymphocytes in rats.Methods:20 clean grade Sprague Dawley(SD)male rats(2 months)were chosen and splenic lymphocytes were isolated aseptically.They were divided into control group and experimental group(I-XIII),which added in the lymphocytes respectively 0,0.1,0.2,0.4,0.8,1,2,4,8,10,20,40,80 and 100 mmol/L boric acid for 24 h,determined by MTT,and Cell Counting Kit-8 method are used to detect boron cytotoxic effects,and flow cytometry to detect lymphocyte proliferation and apoptosis.Results:Cytotoxicity test results can be seen that,compared with the control group,the inhibition rate was significantly decreased in the experimental group III(P<0.05),the proliferation rate was significantly increased(P<0.05),but the inhibition rate of experimental group X-XIII was significantly increased(P<0.01)with the proliferation rate was significantly decreased(P<0.01).Flow cytometry results can be seen that,compared with the control group,the proliferation rate of experimental group III was significantly increased(P<0.01),and the apoptosis rate was significant decreased(P<0.05),and the proliferation rate of experimental group XI and XII were significantly decreased(P<0.01)with the apoptosis rate were significantly increased(P<0.01).Conclusion:Adding 0.4 mmol/L of boric acid is no cytotoxic effect on lymphocytes,It can significantly stimulate lymphocyte proliferation and decrease the inhibition rate of lymphocytes.When adding 20 mmol/L of boric acid,it has significantly cytotoxic effects on lymphocytes and promoting the apoptosis of lymphocytes.