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目的:探讨宣白承气汤对脓毒症大鼠保护作用的细胞信号转导机制。方法:SD大鼠30只,随机分为3组,模型组、假手术组和治疗组各10只。实验期间大鼠禁食不禁水。模型组和治疗组采用盲肠结扎穿刺法,假手术组仅翻动盲肠,不做穿刺结扎处理。治疗组在CLP后8h、12h和24h时宣白承气汤剂按1ml/100g体质量灌胃。第48h时(即实验结束时)心脏取血3ml离心后去上清液测定TNFα、IL-6含量。并取距小肠回盲部上端15cm处的肠管2cm,称重后按每1g湿重加入4℃的PBS10ml比例,冰浴匀浆约1min后,4℃放置4500转/min离心30min后取上清备用。免疫印迹法检测小肠组织中p-p38、total-p38丝裂原活化蛋白激酶表达水平。结果:模型组的TNFα及IL-6浓度与假手术组比较,均明显升高(P<0.05);与模型组比较,治疗组可明显降低血清中TNFα及IL-6浓度(P<0.05)。模型组p-p38/p-38灰度比值为0.48±0.09,假手术组的灰度比为0.16±0.03,治疗组的灰度比为0.21±0.05。与假手术组比较,模型组灰度比值明显增加(P<0.05);与模型组比较,治疗组的灰度比值明显降低(P<0.05)。结论:宣白承气汤保护肠源性脓毒症大鼠的作用机制与抑制小肠组织中p38丝裂原活化蛋白激酶信号通路有关,进而减少炎症介质TNFα、IL-6释放。
Objective: To investigate the protective effect of Xuanbai Chengqi Decoction on septic rats by cell signal transduction mechanism. Methods: Thirty SD rats were randomly divided into three groups: model group, sham operation group and treatment group. Fasting rats can not help but water during the experiment. Cecal ligation and puncture were used in the model group and the treatment group, while the sham operation group only turned the cecum without puncture and ligation. The treatment group at 8h, 12h and 24h after CLP, declare that the gas-bearing decoction by 1ml / 100g body weight gavage. At 48h (ie, at the end of the experiment), 3ml of blood from the heart was centrifuged to determine the contents of TNFα and IL-6 in the supernatant. And from the upper end of the ileum of the small intestine 15cm at the bowel 2cm, after weighing 1g wet weight by adding 4 ℃ PBS10ml ratio, ice bath homogenate for about 1min, 4 ℃ placed 4500 rev / min centrifugation 30min after the supernatant spare. The expression of p-p38, total-p38 mitogen-activated protein kinase in intestinal tissue was detected by Western blotting. Results: The levels of TNFα and IL-6 in model group were significantly higher than those in sham-operated group (P <0.05). Compared with model group, the levels of TNFα and IL-6 in serum of model group were significantly decreased (P <0.05) . The gray-scale ratio of p-p38 / p-38 in model group was 0.48 ± 0.09, that of sham group was 0.16 ± 0.03, and that of the treatment group was 0.21 ± 0.05. Compared with the sham group, the grayscale ratio of the model group increased significantly (P <0.05). Compared with the model group, the grayscale ratio of the treatment group decreased significantly (P <0.05). Conclusion: The mechanism of Xuanbai Chengqi Decoction in protecting gut-derived sepsis rats is related to the inhibition of p38 mitogen-activated protein kinase (MAPK) signaling pathway in small intestine tissues, which in turn reduces the release of inflammatory mediators TNFα and IL-6.