加味清肠解毒汤对溃疡性结肠炎模型大鼠炎症细胞因子及血小板活化功能的影响

发布时间：2018-04-25 19:22

本文选题：溃疡性结肠炎 + 加味清肠解毒汤　；参考：《成都中医药大学》2016年博士论文

【摘要】：1.实验目的通过观察加味清肠解毒汤对TNBS/乙醇复制的溃疡性结肠炎模型大鼠的炎症细胞因子相关指标及血小板活化功能的影响来研究其治疗作用和机制。2.实验方法SPF级SD大鼠60只随机分为空白组、模型组、西药组、中药高剂量组(简称高剂量组)、中药中剂量组(简称中剂量组)、中药低剂量组(简称低剂量组),每组10只。除空白组外,其余采用TNBS/乙醇复制溃疡性结肠炎模型。第一次造模后第5周起给药,连续两周,空白组、模型组给予等体积生理盐水灌胃,西药组给予美沙拉嗪缓释颗粒灌胃,中药各治疗组给予加昧清肠解毒汤水煎液灌胃。给药第7天采血检测TNF-α、TXB2、6-Keto-PGF1α含量；给药第14天采血检测TNF-α、TXB2、6-Keto-PGF1α、PC含量；给药2周后处死所有动物,分别进行结肠组织病理学评分,免疫组化法检测结肠组织IL-1β、IL-6、IL-8的表达,酶联免疫法检测结肠组织IL-10的表达,以及PCR检测结肠组织TLR4、NF-κB的基因表达。3.实验结果(1)采用TNBS/乙醇复制大鼠溃疡性结肠炎模型,造模后实验动物出现典型的腹泻、血便、体重减轻、精神萎靡等表现,经治疗后上述情况逐渐好转；与模型组比较,各治疗组体重恢复有统计学差异(P0.01,P0.05)；(2)与空白组比较,模型组结肠组织损伤符合典型的UC特征,病理学评分显著升高(P0.01)；与模型组比较,各治疗组病理学评分明显下降(P0.01,P0.05);(3)与空白组相比,模型组大鼠结肠组织IL-1β表达明显升高(P0.01)；与模型对照组相比,各治疗组大鼠结肠组织IL-1β表达降低,(P0.05,P0.01);(4)与空白组相比,模型组大鼠结肠组织IL-6表达明显升高(P0.01)；与模型组相比,各治疗组大鼠结肠组织IL-6表达降低(P0.05)；(5)与空白组相比,模型组大鼠结肠组织IL-8表达明显升高(P0.01)；与模型组相比,各治疗组大鼠结肠组织IL-8表达降低(P0.05,P0.01)；(6)空白组不同时间点之间比较,TNF-α的表达无明显差异(P0.05);不同时间点模型组TNF-α的表达较空白组明显增多(P0.01)；与模型组比较,不同时间点除低剂量组之外其余治疗组的TNF-α的表达下调(P0.01,P0.05)；(7)与空白组相比,模型组大鼠结肠组织IL-10含量明显降低(P0.01)；与模型组相比,各治疗组除低剂量组外IL-10的含量有所升高(P0.05,P0.01)；(8)与空白组比较,模型组大鼠结肠组织TLR4、NF-κB的mRNA表达增强(P0.01)；与模型组比较,西药组和中药组大鼠结肠组织中的TLR4、NF-κB的mRNA表达下降(P0.01)；西药组与中药组比较无显著差异(P0.05);(9)与空白组比较,模型组大鼠PC显著升高(P0.01)：与模型组比较,各治疗组PC有所下降,其中西药组、中药高剂量组有统计学差异(P0.05)；(10)空白组组内不同时间点比较,血浆TXB2含量无明显差异(P0.05)；与空白组相比,模型组不同时间点血浆TXB2含量显著升高(P0.01);与模型组比较,除低剂量组外,其余各组不同时间点血浆TXB2含量下降(P0.01,P0.05);(11)空白组组内各时间点比较血浆6-Keto-PGF1α含量无明显差异(P0.05);与空白组比较,模型组不同时间点血浆6-Keto-PGF1α含量升高(P0.05)；与模型组比较,各治疗组不同时间点血浆6-Keto-PGF1α含量升高(P0.05,P0.01)；(12)空白组组内不同时间点比较,TXB2/6-Keto-PGF1α比值无明显差异(P0.05)；与空白组比较,模型组不同时间点TXB2/6-Keto-PGF1α比值显著升高(P0.01)；与模型组比较,各治疗组不同时间点TXB,/6-Keto-PGF1α比值降低(P0.01,P0.05)：4.实验结论(1)加味清肠解毒汤具有改善实验性溃疡性结肠炎组织病理学评分,改善一般情况,促进实验大鼠体重恢复的作用；(2)加味清肠解毒汤降低促炎因子IL-1β、IL-6、IL-8、TNF-α的过度表达,升高抗炎因子IL-10的表达,可能是其治疗TNBS/乙醇复制的大鼠溃疡性结肠炎的机制之一；(3)加味清肠解毒汤治疗实验性溃疡性结肠炎的机制可能与通过抑制TLR4、 NF-κB的过度表达继而抑制下游炎症因子的释放发挥抗炎作用有关；(4)加味清肠解毒汤治疗实验性溃疡性结肠炎的机制可能与通过下调血浆TXB2水平,上调6-Keto-PGF1α含量,降低TXB2/6-Keto-PGF1α比值,降低血小板过度活化有关。
[Abstract]:1. to observe the effect of Jiawei Qing Chang Jiedu soup on the inflammatory cytokine related index and platelet activation function of TNBS/ ethanol replicating ulcerative colitis model rats, and to study its therapeutic effect and mechanism.2. experimental method SPF grade SD rats randomly divided into blank group, model group, western medicine group, high dose group of traditional Chinese medicine (Jane) The high dose group (high dose group), medium dose group (medium dose group), low dose group of traditional Chinese medicine (low dose group), 10 rats in each group. Except for the blank group, the rest used TNBS/ ethanol to copy ulcerative colitis model. After the first model, the drug was given for two weeks, the blank group was given the same volume of normal saline, and the western medicine group was given Meisha. TNF- alpha, TXB2,6-Keto-PGF1 alpha content was measured for seventh days, and the content of TNF- a, TXB2,6-Keto-PGF1 A and PC was detected by blood for fourteenth days; all animals were killed after 2 weeks of administration, and the colon histopathological score and immunohistochemical method were carried out for 2 weeks. The expression of IL-1 beta, IL-6 and IL-8 in colon tissue was measured, the expression of IL-10 in colon tissue was detected by ELISA, and PCR was used to detect TLR4 in colon tissue and the results of NF- kappa B gene expression.3. experimental results (1) the rat model of ulcerative colitis was replicated by TNBS/ ethanol. After the model, the experimental animals showed typical diarrhea, blood stool, weight loss and mental malaise. After treatment, the above conditions gradually improved; compared with the model group, the weight recovery of the treatment groups was statistically different (P0.01, P0.05). (2) compared with the blank group, the damage of the colon tissue in the model group was conformed to the typical UC characteristics, and the pathological score was significantly higher (P0.01); and compared with the model group, the pathological scores of the treatment groups were significantly decreased (P0.01, P0.05). 3) the expression of IL-1 beta in the colon tissue of the model group was significantly higher than that in the blank group (P0.01). Compared with the model control group, the expression of IL-1 beta in the colon tissue of the rats in the treatment group was reduced, (P0.05, P0.01). (4) the expression of IL-6 in the colon tissue of the model group was significantly higher than that in the blank group (P0.01). Compared with the model group, the colon group of the rats in the treatment group was compared with the model group. The expression of IL-6 was reduced (P0.05); (5) the expression of IL-8 in the colon tissue of the model group was significantly higher than that in the blank group (P0.01). Compared with the model group, the expression of IL-8 in the colon tissue of the rats in the treatment group decreased (P0.05, P0.01), and (6) there was no significant difference in the expression of TNF- a (P0.05) in the blank group at different time points (P0.05), and the model group of TNF- a at different time points. Compared with the model group, the expression of TNF- alpha in the other treatment groups was down down (P0.01, P0.05) at different time points except the low dose group. (7) the IL-10 content in the colon tissue of the model group was significantly lower than that in the blank group (P0.01). Compared with the model group, the content of IL-10 in the treatment group except the low dose group was compared with the model group. (P0.05, P0.01); (8) compared with the blank group, the colon tissue of the rats in the model group was TLR4, and the mRNA expression of NF- kappa B was enhanced (P0.01). Compared with the model group, the mRNA expression of NF- kappa B in the colon tissue of the western medicine group and the Chinese medicine group was decreased (P0.01); (9) compared with the blank group, the model was compared with the blank group. Group PC significantly increased (P0.01): compared with the model group, PC decreased in the treatment group, and there was a statistical difference between the western medicine group and the high dose group (P0.05). (10) there was no significant difference in plasma TXB2 content in the blank group at different time points (P0.05). Compared with the empty white group, the plasma TXB2 content was significantly increased in the model group at different time points (P0.0). 1): compared with the model group, the plasma TXB2 content decreased (P0.01, P0.05) at different time points except the low dose group, and (11) there was no significant difference in plasma 6-Keto-PGF1 alpha content in the blank group at all time points (P0.05). Compared with the blank group, the plasma level of 6-Keto-PGF1 alpha was increased (P0.05) in the model group (P0.05), and the treatment was compared with the model group. The plasma 6-Keto-PGF1 alpha content increased at different time points in the treatment group (P0.05, P0.01). (12) there was no significant difference in the ratio of TXB2/6-Keto-PGF1 a (P0.05) at different time points in the blank group (P0.05). Compared with the blank group, the TXB2/6-Keto-PGF1 alpha ratio in the model group increased significantly at different time points (P0.01). Compared with the model group, the different time points of the treatment groups were TXB, /6. The -Keto-PGF1 alpha ratio decreased (P0.01, P0.05):4. experimental conclusion (1) Jiawei Qing Chang Jiedu Decoction can improve the histopathological score of experimental ulcerative colitis, improve the general situation, promote the effect of weight recovery of experimental rats, and (2) the overexpression of IL-1 beta, IL-6, IL-8, TNF- a, and increase the anti inflammatory factors of the IL-1 beta, IL-6, IL-8, TNF- alpha in the proinflammatory cells. The expression of IL-10 may be one of the mechanisms for the treatment of TNBS/ ethanol replicating ulcerative colitis. (3) the mechanism of the treatment of experimental ulcerative colitis in the treatment of experimental ulcerative colitis may be related to the inhibition of the overexpression of TLR4, NF- kappa B and the inhibition of the release of the downstream inflammatory factors; (4) the flavored clearing intestine detoxification soup The mechanism of treating experimental ulcerative colitis may be related to the reduction of plasma TXB2 level, the increase of 6-Keto-PGF1 alpha content, the reduction of the ratio of TXB2/6-Keto-PGF1 a, and the reduction of platelet activation.

【学位授予单位】：成都中医药大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R259;R-332

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