活血通腑方对术后腹腔粘连大鼠肠黏膜免疫屏障功能的影响
Effects of Classical Prescriptions of Blood-activating and Organ-purging Formula on Immunologic Barrier Function of Intestinal Mucosa in Rats with Postoperative Peritoneal Adhesion
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摘要: 目的 观察活血通腑方对术后腹腔粘连早期肠道黏膜屏障的影响。方法 取SD雄性大鼠60只,随机分为对照组、模型组、活血通腑方组,除对照组外,其余动物均采用锉刀法构建腹腔粘连大鼠模型,分别于术后第3、6、12 h和24 h给予相应剂量的药物,术后48 h采血及组织样本,ELISA法测定血浆中TNF-α水平,免疫组化法检测肠黏膜sIgA的含量和CD4细胞数量,TUNEL法检测肠黏膜下淋巴细胞凋亡情况。结果 与对照组相比,6、12 h和24 h后模型组血浆中TNF-α水平具有统计学差异(P<0.05~0.01);3、6 h模型组黏膜sIgA表达明显下降;肠黏膜下淋巴细胞凋亡明显(P<0.01),12、24 h时sIgA的分泌量明显增多(P<0.05);而模型组12、24 h与3、6 h相比,黏膜下淋巴细胞凋亡明显下降接近对照组水平;模型组3 h肠道黏膜CD4细胞数量明显下降(P<0.01);6、12、24 h时CD4细胞数量3组比较未见明显异常(P>0.05)。活血通腑方在术后3 h可显著降低模型鼠血浆中TNF-α水平;增高肠道黏膜sIgA含量和CD4细胞数量;减弱黏膜下淋巴细胞凋亡。结论 活血通腑方组可通过降低血TNF-α的水平,减轻黏膜下淋巴细胞凋亡,增加肠道黏膜sIgA含量和CD4细胞数量,提高局部免疫水平,抑制炎症反应。Abstract: OBJECTIVE To explore the effect of blood-activating and organ-purging formula in vivo of intestinal mucosal barrier after surgery in peritoneal adhesion. METHODS 60 male SD rats were randomly divided into control group, model group and Huoxue Tongfu Decoction group. Besides the control group, the rest of the rats were established with grater method, and the same dose of drugs were given at 3, 6, 12, 24 h after operation. The blood and tissue samples were taken after 48 h. TNF-α levels in plasma were determined by ELISA, sIgA in intestinal mucosa was detected by immunohistochemical method and the number of content of CD4 cells, lymphocytes apoptosis of intestinal mucosa were detected by TUNEL. RESULTS Compared with the control group, the plasma levels of TNF-α in the model group of 6, 12, 24 h showed significant difference (P<0.05~0.01), intestinal mucosa under lymphocyte were apoptosis significantly (P<0.01), 12 h and 24 h secretion IgA significantly increased (P<0.05), and the model group 12 h, 24 h and 3 h, 6 h, submucous lymphocyte apoptosis decreased close to the level of control group, model group 3 h CD4 in intestinal mucosa cell number decreased significantly (P<0.01), there was no significant difference in the number of CD4 cells when three groups at 6 h, 12 h and 24 h (P>0.05). Prescription in 3 h after operation can significantly reduce the levels of TNF-α in plasma of model rats increased number of intestinal mucosae, the content of IgA and CD4 cells decreased, submucous lymphocyte apoptosis. CONCLUSION Blood-activating and organ-purging formula group can reduce the blood level of TNF- alpha and submucous lymphocyte apoptosis, increase the number of intestinal mucosal s content of IgA and CD4 cells, improving local immunity, inhibiting the inflammatory reaction.
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