【摘要】 目的探讨黄芪(AM)对新生儿脐血淋巴细胞凋亡的影响及其作用机制。方法随机选择30例正常足月新生儿为研究对象,收集并分离脐血单个核细胞(CBMC),体外分别经植物血凝集素(PHA)、PHA联合白细胞介素6(IL-6)、PHA联合AM刺激培养48 h,采用丫啶橙-溴化乙锭(AB-EO)染色法测定细胞凋亡水平,间接免疫荧光法测定CBMC中CD38阳性和CD25阳性细胞百分率,酶联免疫吸附试验(ELISA)检测培养上清液中IL-6的水平。结果PHA+AM刺激组细胞凋亡率明显低于单纯PHA刺激组(F=205.91,q=25.16,P<0.001),PHA+IL-6刺激组明显低于单纯PHA刺激组(q=24.54,P<0.001),而PHA+IL-6刺激组与PHA+AM刺激组间细胞凋亡水平比较差异无显著性(q=0.62,P>0.05)。PHA+AM刺激组CD38阳性细胞百分率明显低于单纯PHA刺激组(F=790.12,q=48.95,P<0.001),PHA+IL-6刺激组也明显低于单纯PHA刺激组(q=48.42,P<0.001),但PHA+IL-6刺激组与PHA+AM刺激组间比较差异无显著性(q=0.53,P>0.05)。PHA+AM刺激组CD25阳性细胞百分率明显高于单纯PHA刺激组(F=221.38,q=26.60,P<0.001),PHA+IL-6刺激组也明显高于单纯PHA刺激组(q=24.86,P<0.001),但PHA+IL-6刺激组与PHA+AM刺激组间比较差异无显著性(q=1.74,P>0.05)。PHA+AM刺激组培养上清液中IL-6水平明显高于单纯PHA刺激组(t′=6.50,P<0.001)。CBMC经48h培养后,凋亡细胞百分率与培养上清液中IL-6浓度呈明显负相关(r=-0.75,P<0.05)。结论黄芪能明显抑制CBMC体外经PHA激活后的细胞凋亡,其机制可能与促进CBMC中胸腺细胞向T淋巴细胞分化、增强脐血淋巴细胞活化和增加IL-6产生水平等有关。更多还原

【Abstract】 ObjectiveTo study the effects of Astragalus Membranaceus（AM） on neonatal lymphocyte apoptosis in cord blood and to study the their mechanism.MethodsThirty full-term neonates were enrolled in this study.Cord blood mononuclear cells（CBMC） were cultured in vitro with pure PHA,PHA combined with IL-6 or AM,respectively,for 48 hours.The apoptosis indexes of lymphocytes of CBMC after cultivation were determined by AB-EO dying method.The positive expression of CD₃₈ antigen and CD₂₅ antigen were detected by indirect immunofluorescence procedure.The levels of IL-6 in the supernatants of CBMC was detected by ELISA.ResultsThe apoptosis index（AI） of the PHA combined with AM group and the PHA combined with IL-6 group were both lower than that of the pure PHA group（F=205.91;q=25.16,24.54;P<0.001）,but there was no difference between the AI of the PHA combined with IL-6 group and that of the PHA combined with AM group（q=0.62,P>0.05）.The positive expression of CD₃₈ of the PHA combined AM group and the PHA combined IL-6 group were all lower than that of the pure PHA group（F=790.12;q=48.95,48.42;P<0.001）,but there was no difference between the positive expression of CD₃₈ of PHA combined with IL6 group and that of PHA combined with AM group（q=0.53,P>0.05）.The positive expression of CD₂₅ of the PHA combined AM group was higher than that of the pure PHA group（F=221.38,q=26.60,P<0.001）,and the positive expression of CD₂₅ of the PHA combined IL-6 group was higher than that of the pure PHA group（q=24.86,P<0.001）.But there was no difference between the positive expression of CD₂₅ of the PHA combined with IL-6 group and that of the PHA combined with AM group（q=1.74,P>0.05）.The IL-6 level in supernatants of CBMC of the PHA combined with AM group was significantly higher than that of the pure PHA group（t′=6.50,P<0.001）.Cultured for 48 hours,the AI of CBMC had negative correlation with the level of IL-6 of the supernatants（r=-0.75,P<0.05）.ConclusionAM can significantly inhibit the apoptosis of CBMC after PHA stimulation in vitro.The possible mechanism may be that AM can increase the cell differentiation from thymocytes to T lymphocytes,enhance the activation of cord blood lymphocytes and increase the production of IL-6 level.更多还原