翁连解毒汤对溃疡性结肠炎大鼠IL-17、TLR2的影响

发布时间：2018-03-06 08:50

  本文选题：溃疡性结肠炎　切入点：翁连解毒汤　出处：《河北医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:溃疡性结肠炎(ulcerative colitis,UC)是发生在直肠和结肠的、累及黏膜及黏膜下层的慢性非特异性炎症性疾病,发病率高,且易复发,病程较长。本实验通过观察翁连解毒汤方对溃疡性结肠炎大鼠的一般状态、疾病活动指数(DAI)评分、血清IL-17含量及结肠组织TLR2表达的影响,探讨翁连解毒汤治疗溃疡性结肠炎的作用机制,为临床应用提供实验依据。方法:将60只洁净雄性健康Wistar大鼠根据随机数字表随机分为6组,每组10只,分别为空白对照组(NG)、模型组(MG)、西药美沙拉嗪组(XG)、中药高剂量组(HG)、中药中剂量组(ZG)、中药低剂量组(LG)。空白对照组(NG)正常饲养,其余5组大鼠均采用三硝基苯磺酸(TNBS)/乙醇联合造模方法建立溃疡性结肠炎大鼠模型。自造模后第4天,各组大鼠除NG、MG正常饲养外,余治疗组大鼠除基础饲养外合用药物灌胃,其中XG组大鼠采取美沙拉嗪混悬液灌胃,HG、ZG和LG大鼠采用不同的浓度的翁连解毒汤灌胃治疗。连续治疗2周后,观察大鼠在各个时期的精神状态、饮食、被毛、粪便性状、体重等一般状态的变化并记录,做出疾病活动指数评分表。治疗结束后,各组大鼠禁食不禁水24小时,利多卡因麻醉大鼠取股动脉血,用双抗体夹心ABC-ELLISA法检测血清IL-17含量,解剖并取其肉眼观察病变较重部位的结肠组织,常规石蜡包埋、切片、HE染色,检测TLR2的表达情况。结果:1一般情况观察:NG大鼠一般状态良好,余组大鼠精神萎靡,反应迟钝,进食水量减少,体重下降,粪便稀薄夹杂粘液、脓血。药物灌胃治疗后各组大鼠精神状态好转,进食量明显增多,体重下降减慢,粘液、脓血便减少或消失,其中HG改善最明显,ZG、XG改善无明显差异,较HG略差,较LG较好。DAI评分结果提示:XG、HG、ZG、LG与MG相比较低,差异有统计学意义(P0.05);XG、ZG、LG与HG相比较高,差异有统计学意义(P0.05);XG与ZG无统计学意义(P0.05);XG、ZG与LG相比较低,差异有统计学意义(P0.05)。2结肠黏膜肉眼与病理学观察肉眼观察:NG大鼠上皮黏膜光滑完整。MG大鼠黏膜充血水肿糜烂面最明显。HG大鼠黏膜较光滑,溃疡面极少。XG和ZG大鼠黏膜欠光滑,可见较少的溃疡面。LG大鼠黏膜组织轻度充血水肿,溃疡灶较多。组织病理学观察:NG大鼠黏膜光滑,腺体结构排列规则。MG大鼠黏膜粗糙,可见多发、弥漫性糜烂或溃疡面,局部腺体破坏最明显,可见大量炎性细胞浸润。HG组黏膜较光滑完整,新生腺体最多。XG和ZG大鼠黏膜欠光滑,可见少许炎性细胞浸润,溃疡面较少。LG大鼠黏膜组织轻度充血水肿,遗留溃疡灶较多,少数处于愈合期,溃疡面及边缘覆于少量新生腺体,见附图Fig.1-Fig.12。3指标检测IL-17含量比较:MG、XG、HG、ZG、LG与NG相比含量较高,差异有统计学意义(P0.05);HG、XG、ZG、LG与MG相比,含量较低,差异有统计学意义(P0.05);XG、ZG、LG与HG相比含量较高,差异有统计学意义(P0.05);XG、ZG与LG相比含量较低,且差异有统计学意义(P0.05);XG与ZG相比较,差异无统计学意义(P0.05)。TLR2表达比较:MG、XG、HG、ZG、LG与NG相比表达较高,差异有统计学意义(P0.05);XG、HG、ZG、LG与MG相比,表达较低,差异具有统计学意义(P0.05);HG较XG、ZG、LG相比,表达较低,差异有统计学意义(P0.05);ZG、XG与LG相比,表达较低,差异有统计学意义(P0.05);XG与ZG相比较,差异无统计学意义(P0.05)。结论:翁连解毒汤能够改善溃疡性结肠炎大鼠的一般情况并且能够修复损伤的黏膜,降低DAI评分,并呈现一定的量-效关系,其特点为随着中药浓度的升高而疗效增加,其作用机制可能通过降低TLR2的表达、减少IL-17的含量,从而阻断炎症反应,减少炎性细胞浸润,刺激局部组织再生与修复,达到愈合溃疡面的作用。
[Abstract]:Objective: ulcerative colitis (ulcerative colitis, UC) occurred in the rectum and colon, involving the mucosa and submucosa of chronic non-specific inflammatory disease, high incidence and recurrence, longer duration. This experiment through the observation of Weng Jiedu Decoction on even ulcerative colitis rats general state, disease the activity index (DAI) score, the expression of TLR2 IL-17 in serum and colon tissue, to explore the mechanism of treating ulcerative colitis even Weng Jiedu Decoction, provide experimental basis for clinical application. Methods: 60 clean healthy male Wistar rats according to randomly divided into 6 groups, 10 rats in each group, respectively. For the control group (NG), model group (MG), western medicine group (XG) the beauty of salad, high dose of Chinese medicine group (HG), Chinese medicine dose group (ZG), low dose of Chinese medicine group (LG). The blank control group (NG) of normal feeding, the remaining 5 groups of rats were three nitrobenzene sulfonic acid (TNBS) / ethanol combined modeling method to establish the rat model of ulcerative colitis. The model was made after fourth days, the rats in NG, MG more than normal feeding, the rats in the treatment group in addition to the basic drug fed by gavage, the rats in the XG group to take the salad suspension gavage, HG ZG, and LG of rats with different concentrations of Weng even Jiedu Decoction by gavage. After 2 weeks of continuous treatment, rats were observed in each period of the mental state, diet, hair, stool, weight change etc. the general state and make record, disease activity index score. After treatment, fasting rats can not help water for 24 hours, lidocaine anesthesia rats from femoral artery blood, by double antibody sandwich ABC-ELLISA assay of serum IL-17 content, and the anatomy of the colon tissue observed lesions heavier parts, paraffin embedded sections, HE staining to detect the expression of TLR2. Results: 1 the general situation Observe the general condition of rats: NG rats were more than good, apathetic, unresponsive, eating less water, weight loss, thin stool with mucus, pus and blood. Drug irrigation improved mental state of rats after the treatment of stomach, food intake increased significantly, decreased body weight decreased, mucous pus blood decreased or disappeared. The most obvious improvement in HG, ZG, XG improvement had no significant difference compared with HG slightly worse than LG better.DAI score showed: XG, HG, ZG, LG and MG are relatively low, the difference was statistically significant (P0.05); XG, ZG, LG and HG are relatively high, the difference was statistically significant (P0.05); XG and ZG had no statistical significance (P0.05); XG, ZG and LG are relatively low, the difference was statistically significant (P0.05) were observed with naked eye and eye pathology of.2 in colonic mucosa of NG rats mucosa smooth and complete the most obvious.MG rat mucosal edema.HG rat mucosal erosion surface was relatively smooth. The ulcer rarely.XG and ZG rats Mucosa less smooth, less visible mucosa ulcer.LG rats with mild hyperemia and edema, ulcer more. Histopathological observation: mucosal NG rats smooth, glandular structures arranged in mucosa of.MG rats of rough rules, there are multiple, diffuse erosion or ulcer, local glands broken the most obvious, visible inflammatory infiltration of inflammatory cells in group.HG mucosa is smooth and complete, the new maximum of.XG and ZG in rat glandular mucosa was not smooth, there is little inflammatory cell infiltration in mucosa of rats with ulcer less.LG mild hyperemia and edema, ulcer left more, a few in the healing period, the ulcer surface and edge to cover a few new glands, see figure Fig.1-Fig.12.3 index to detect the content of IL-17: MG, XG, compared with HG, ZG, LG were higher compared with NG, the difference was statistically significant (P0.05); HG, XG, ZG, LG compared with MG, content is low, the difference was statistically significant (P0.05); XG, ZG, LG and HG were compared 杈冮珮,宸�寮傛湁缁熻�″�︽剰涔�(P0.05);XG,ZG涓嶭G鐩告瘮鍚�閲忚緝浣�,涓斿樊寮傛湁缁熻�″�︽剰涔，

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