枸杞多糖抑制过氧化氢诱导的子宫内膜间质细胞的凋亡作用

Inhibiting effects of Lycium Barbarum Polysaccharides on the apoptosis of human endometrial stromal cells induced by hydrogen peroxide

目的通过采用过氧化氢(H_2O_2)诱导人子宫内膜间质细胞(ESCs)凋亡,探讨经过枸杞多糖(LBP)干预后对其凋亡的抑制作用及其机制的研究。方法体外分离、培养ESCs,将其分为4组:正常组:常规培养;H_2O_2组:培养液中加入H_2O_2;LBP组:培养液中加入100μg/ml LBP;LBP+H_2O_2组:LBP先作用1 h后再加入H_2O_2共培养30 min。流式细胞仪分析各组细胞的凋亡率;用比色法检测细胞培养液中丙二醛(MDA)的含量和超氧化物歧化酶(SOD)的活性;免疫细胞化学法检测细胞质中Bax和bcl-2的水平。结果与正常组相比,H_2O_2组细胞凋亡率增加(50.35±1.83),MDA的含量升高[(8.67±0.63)μmol/L]而SOD的活性降低[(2.09±0.59)KU/g Pro],细胞中Bax表达上调(60.45±7.63)而bcl-2的表达下调(23.59±2.67),其差异有统计学意义(P<0.05);与H_2O_2组相比,LBP+H_2O_2组细胞凋亡率明显降低(34.63±1.32),MDA含量下降[(4.58±0.41)μmol/L],SOD活性上升[(3.36±0.41)KU/g Pro],细胞中Bax表达下调(30.65±5.42)而bcl-2的表达上调(53.42±6.39),其差异有统计学意义(P<0.05)。结论LBP对H_2O_2导致的ESCs凋亡有抑制作用,其机制是通过提高ESCs中SOD的活性而降低MDA的含量、上调bcl-2并下调Bax的表达水平。

Objective]To investigate apoptosis inhibition function and mechanism of Lycium Barbarum Polysaccharides（LBP）on human endometrial stromal（ESCs）which were induced by hydrogen peroxide（H2O2）.[Methods]ESCs were isolated and cultured in vitro,and divided into four groups：normal group：ESCs were cultured with regular method;H2O2 group：ESCs were cultured with H2O2;LBP group：ESCs were cultured with 100μg/ml LBP;LBP＋H2O2 group：ESCs were cultured with LBP for one hour and then co-cultured with LBP and H2O2.Cell apoptosis rate was analyzed by Flow cytometry.The contents of malondiadehyd e（MDA）and the activity of superoxide dismutase（SOD）were measured by using colorimetry method.The protein expression levels of Bcl-2 and Bax in the cytoplasm were detected by.immunocytochemistry method.[Results]Compared with normal group,the cell apoptosis rate increased（50.35±1.83）in H2O2 group,the level of MDA increased[（8.67±0.63）μmol/L）]while the activity of SOD decreased[（2.09±0.59）k U/g Pro],the expression level of Bax increased（60.45±7.63）while the expression level of Bcl-2 decreased（23.59±2.67）,and the difference was statistically significant（P〈0.05）.Compared with H2O2 group,the cell apoptosis rate decreased（34.63±1.32）in LBP＋H2O2 group,the level of MDA decreased[（4.58±0.41）μmol/L]while the activity of SOD increased[（3.36±0.41）k U/g Pro],the expression level of Bax decreased（30.65±5.42）while the expression level of Bcl-2（53.42±6.39）,and the difference was statistically significant（P〈0.05）.[Conclusion]LBP can suppress the ESCs apoptosis induced by H2O2,and its mechanism is that LBP can up-regulate the activity of SOD and the protein expression of Bcl-2,down-regulate the content of MDA and protein expression of Bax.