黄芪对脓毒症大鼠肠道相关淋巴细胞凋亡的影响

发布时间：2018-11-14 14:27

【摘要】：目的：观察黄芪对脓毒症大鼠肠道相关淋巴细胞凋亡的影响。方法：选择使用盲肠结扎并穿孔术复制脓毒症大鼠模型，将健康雄性SD大鼠72只（25020）g，随机分为3组，假手术组、黄芪治疗组和脓毒症组，在造模成功后的12h、24h、72h三个时间点分别留取肠集合淋巴结(PP结）、回盲部上段小肠及小肠粘液灌洗液，采用TUNEL检测PP结淋巴细胞的凋亡率；ELISA测定肠灌洗液IgA含量；免疫组化法观察肠固有层sIgA阳性细胞数量的变化以及PP结内淋巴细胞Caspase-3蛋白的阳性表达情况。结果： 1、PP结淋巴细胞凋亡率假手术组在12h、24h及72h各时间点PP结内淋巴细胞均存在较低的凋亡率无显著差异（P＞0.05）；脓毒症组及黄芪治疗组在12h、24h、72h各时间点的PP结淋巴细胞凋亡率较假手术组均升高（P＜0.05），12h最高，此后逐步降低，72h最低但较假手术组仍明显升高（P＜0.05），且在各时间点脓毒症组较黄芪治疗组也有所升高（P＜0.05），差异有统计学意义。 2、肠固有层sIgA阳性细胞数量的变化假手术组在12h、24h及72h三个时间点的肠固有层sIgA阳性细胞数量变化无显著差异（P＞0.05）；脓毒症组及黄芪治疗组在12h、24h、72h各时间点的肠固有层sIgA阳性细胞数量较假手术组均减少（P＜0.05），12h最少，此后逐步增加，72h最多与假手术组对比仍减少（P＜0.05），且在各时间点脓毒症组的肠固有层sIgA阳性细胞数量也少于黄芪注射液治疗组（P＜0.05），差异有统计学意义。 3、PP结内淋巴细胞Caspase-3蛋白的阳性表达情况假手术组在12h、24h和72h各个时间点的PP结淋巴细胞Caspase-3蛋白均有较低的阳性表达，各时间点的积分统计无统计学意义（P＞0.05）；脓毒症组及黄芪治疗组在12h、24h及72h各时间点的PP结淋巴细胞Caspase-3蛋白阳性表达的积分与假手术组对比，，均显著增高(P＜0.05)，12h最高，此后逐步降低，72h最低但仍高出数倍（P＜0.05），且在各时间点脓毒症组较黄芪治疗组增高更加明显（P＜0.05），差异有统计学意义。 4、肠灌洗液IgA水平的变化假手术组在12h、24h及72h三个时间点的肠灌洗液IgA浓度比较无显著差异（P＞0.05）；脓毒症组及黄芪治疗组在12h、24h、72h各时间点的肠IgA水平均显著低于假手术组（P＜0.05），12h最低，此后逐步升高，72h最高与假手术组对比仍明显下降（P＜0.05），且在各时间点脓毒症组较黄芪治疗组也有所降低（P＜0.05），差异有统计学意义。结论：黄芪能有效抑制脓毒症大鼠肠道相关淋巴细胞的过度凋亡，保护肠道免疫屏障，具有改善脓毒症大鼠的肠道黏膜免疫功能。
[Abstract]:Objective: to observe the effect of astragalus on apoptosis of intestinal associated lymphocytes in septic rats. Methods: sepsis rat model was induced by cecal ligation and perforation. 72 healthy male SD rats (25020) g were randomly divided into 3 groups: sham operation group, astragalus membranaceus treatment group and sepsis group. At 72 h, the PP nodes were collected respectively, and the upper ileocecal small intestine and mucus lavage fluid were used to detect the apoptosis rate of PP node lymphocytes by TUNEL. ELISA was used to determine the content of IgA in the intestinal lavage fluid, and the changes of sIgA positive cells in the lamina propria and the expression of Caspase-3 protein in the PP node were observed by immunohistochemical method. Results: 1 there was no significant difference in apoptosis rate of lymphocytes in PP knots between 12h and 72h in sham operation group (P > 0. 05). The apoptosis rate of PP knots in sepsis group and astragalus membranaceus treatment group was higher than that in sham-operated group at 24 h and 72 h (P < 0. 05), the highest at 12 h, and then decreased gradually. The lowest value in 72 hours was higher than that in sham operation group (P < 0.05), and the sepsis group was higher than that of Astragalus membranaceus group at every time point (P < 0.05), and the difference was statistically significant. 2, the change of the number of sIgA positive cells in lamina propria of intestine; there was no significant difference in the number of sIgA positive cells in the lamina propria of intestine at 24 and 72 hours after sham-operation (P > 0. 05). The number of sIgA positive cells in intestinal lamina propria of sepsis group and astragalus membranaceus treatment group at 24 h and 72 h were lower than that in sham operation group (P < 0. 05), and at 12 h, the number of sIgA positive cells in lamina propria was increased gradually. At 72 hours, the number of sIgA positive cells in intestinal lamina propria in sepsis group was lower than that in Huangqi injection group (P < 0. 05). 3The expression of Caspase-3 protein in lymphocytes of PP knots was lower in sham-operated group at 24 h and 72 h after operation, and the expression of Caspase-3 protein in lymphocytes of PP node was lower than that in sham operation group at 24 h and 72 h, respectively. There was no statistical significance in integral statistics at each time point (P > 0.05). In sepsis group and astragalus membranaceus treatment group, the positive expression of Caspase-3 protein in PP node lymphocytes at 12h and 72h was significantly higher than that in sham operation group (P < 0. 05), the highest at 12 h, and then decreased gradually. 72h was the lowest but still several times higher (P < 0.05), and at each time point the sepsis group was higher than the astragalus treatment group (P < 0.05), the difference was statistically significant. 4, the change of IgA level of intestinal lavage fluid in sham-operated group, there was no significant difference in IgA concentration of intestinal lavage fluid at 24 h and 72 h in sham operation group (P > 0. 05). The intestinal IgA levels in sepsis group and astragalus membranaceus treatment group were significantly lower than those in sham operation group (P < 0. 05) at 12 h and 24 h / 72 h, the lowest at 12 h, and then increased gradually. The highest level at 72 h was still significantly lower than that in sham operation group (P < 0. 05). At each time point, sepsis group was lower than that of astragalus treatment group (P < 0.05), the difference was statistically significant. Conclusion: Astragalus membranaceus can effectively inhibit the excessive apoptosis of intestinal lymphocytes and protect the intestinal immune barrier in septic rats. It can improve the intestinal mucosal immune function of septic rats.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R459.7

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