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目的探讨黄芪(astragalusmenbranaceus,AM)对新生儿脐血淋巴细胞凋亡的影响及其机制。方法收集并分离30例正常足月新生儿脐血单个核细胞(cordbloodmononuclearcells,CBMC),分别经(phytohemagglutinin,PHA)、PHA+IL-6和PHA+AM刺激培养48h,采用丫啶橙-溴化乙锭染色法测定细胞凋亡水平,间接免疫荧光法测定CD38+和CD25+细胞百分率,ELISA方法检测培养液上清中IL-6水平。结果(1)PHA+AM、PHA+IL-6和单纯PHA组细胞凋亡率(x±s)分别为(16．5±3．5)％、(16．9±4．0)％和(32．4±2．8)％,PHA+AM和PHA+IL-6组均明显低于单纯PHA组(q=25．16,24．54;P<0．01),而前二者间差异无统计学意义(q=0．62,P>0．05);(2)PHA+AM、PHA+IL-6和单纯PHA组CD38+细胞百分率分别为(9．8±1．8)％、(9．9±0．6)％和(22．3±1．5)％,PHA+AM和PHA+IL6组明显低于单纯PHA组(q=48．95,48．42;P<0．01),但前二者间差异无统计学意义(q=0．53,P>0．05);PHA+AM、PHA+IL-6和单纯PHA组CD25+细胞百分率分别为(83．2±2．4)％、(82．1±3．0)％和(65．8±4．9)％,PHA+AM和PHA+IL-6组明显高于单纯PHA组(q=26．60,24．86;P<0．01),而前二组间比较差异无统计学意义(q=1．74,P>0．05);CBMC中CD38+细胞百分率与细胞凋亡率呈正相关(r=0．68,P<0．01),而CD25+细胞百分率与细胞凋亡率呈负相关(r=-0．65,P<0．01);(3)PHA+AM组培养上清液中IL-6水平[(253．0±30．8)ng／L]明显高于单纯PHA组[(37．9±12．2)ng／L](t=6．50,P<0．01),CBMC凋亡率与培养上清液中IL-6浓度呈负相关(r=-0．75,P<0．01)。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。
Objective To investigate the effect and mechanism of Astragalus usbranaceus (AM) on lymphocyte apoptosis in neonatal cord blood. Methods Cord blood mononuclear cells (CBMCs) from 30 normal term neonates were collected and isolated. The cells were stimulated with phytohemagglutinin (PHA), PHA + IL-6 and PHA + AM for 48h. The level of CD38 + and CD25 + cells was determined by indirect immunofluorescence assay. The level of IL-6 in culture supernatant was detected by ELISA. Results (1) The apoptotic rate (x ± s) of PHA + AM, PHA + IL-6 and PHA group were (16.5 ± 3.5)% and (32.4 ± 2.8)%, PHA + AM and PHA + IL-6 groups were significantly lower than those of PHA alone group (q = 25.16,24.54; P <0.01) (2) The percentage of CD38 + cells in PHA + AM, PHA + IL-6 and PHA group were (9.8 ± 1.8) %, (9.9 ± 0.6)% and (22.3 ± 1.5)% respectively. The PHA + AM and PHA + IL6 groups were significantly lower than those in the PHA group alone (q = 48.95 and 48.42; P (P <0.05). The percentages of CD25 + cells in PHA + AM, PHA + IL-6 and PHA group were (83 .2 ± 2.4%, (82.1 ± 3.0)% and (65.8 ± 4.9)%, respectively. The PHA + AM and PHA + IL- 26.60,24.86; P <0.01), while there was no significant difference between the first two groups (q = 1.74, P> 0.05). The percentage of CD38 + (R = 0.68, P <0.01), while the percentage of CD25 + cells was negatively correlated with apoptosis rate (r = -0.65, P <0.01); (3) IL-6 level in the culture supernatant [(253.0 ± 30.8) ng / L] Ming (37.9 ± 12.2) ng / L] (t = 6.50, P <0.01), the apoptosis rate of CBMC was negatively correlated with the concentration of IL-6 in culture supernatant (R = -0.75, P <0.01). Conclusion Astragalus can significantly inhibit CBMC apoptosis in vitro after PHA activation, and its mechanism may be related to the promotion of CBMC thymocyte differentiation to T lymphocytes, enhance cord blood lymphocyte activation and increase the level of IL-6 production.