【摘要】：目的:本實驗旨在通過愈潰方(YKR)對三硝基苯磺酸(TNBS)誘導的潰瘍性結腸炎大鼠結腸黏膜屏障功能調(diào)節(jié)的作用,通過對緊密連接蛋白Claudin-2和Claudin-7及免疫蛋白MBL和MASP-2蛋白表達情況的研究,探討愈潰方在潰瘍性結腸炎大鼠的腸道粘膜修復功能以及對腸道免疫功能調(diào)節(jié)作用。材料與方法:成年SD級雄性大鼠60只,隨機分配6組,分為A.空白組、B.模型組、C.YKRH劑量組、D.YKRI劑量組、E.YKRL劑量組,F.美沙拉嗪組。除正常對照組外,B、C、D、E、F組大鼠均應用TNBS誘導SD大鼠UC發(fā)病,造模前禁食不禁水24h,灌腸前撫摸大鼠背部刺激遠端大便排空,腹腔注射水合氯醛麻醉后,倒懸位,將直徑0.2cm的聚乙烯管插入結腸8cm處,緩慢灌入TNBS溶液(TNBS為5%水溶液,與無水乙醇按1:1體積配制,乙醇終濃度50%),劑量為60mg/kg,灌腸后倒懸位持續(xù)1min,建模第2天發(fā)現(xiàn)各組模型大鼠均出現(xiàn)稀便及血便情況,開始灌胃給予中藥及美沙拉嗪,A、B組大鼠灌胃給予0.5%羧甲基纖維素鈉,給藥劑量根據(jù)人體轉(zhuǎn)換公式進行轉(zhuǎn)換計算后C、D、E組大鼠灌胃給予不同劑量中藥,中藥高劑量組14mg/g/d,中藥正常劑量組7mg/g/d,中藥低劑量組4mg/g/d給藥劑量,F組大鼠灌胃給予0.2g/kg/d的美沙拉嗪混懸液,分別溶于5ml純凈水中給予灌胃,連續(xù)給藥14天,處死大鼠,對大鼠DAI、CDMI、TDI進行評分,并取結腸標本,采用免疫組化技術,對Claudin-2和Claudin-7及MBL和MASP-2蛋白表達進行檢測,通過統(tǒng)計學軟件進行分析。結果:1.YKR可以有效改善UC大鼠癥狀,在改善大鼠體重、隱血、大便效果上,YKR高劑量組美沙拉嗪組YKR中劑量組YKR低劑量組。TNBS誘導UC大鼠在使用YKR中藥高中劑量后,大鼠DAI、CMDI、TDI評分,效果與美沙拉秦組無統(tǒng)計學意義,均有效降低評分。而YKR低劑量組,大鼠DAI、CMDI、TDI評分與模型組無差別,故無效;2.TNBS誘導SD級大鼠結腸粘膜損傷,相關緊密連接蛋白Claudin-2升高,Claudin-7降低進而影響腸黏膜重建,通透性增加。在使用YKR中高劑量組可使Claudin-2降低,Claudin-7升高,有效改善腸黏膜恢復,低劑量組不明顯,故無效;3.TNBS誘導SD級大鼠結腸可使免疫蛋白MBL蛋白與MASP-2蛋白下降。TNBS誘導SD級大鼠在使用YKR中高劑量組可使MBL蛋白與MASP-2蛋白回升,低劑量組不明顯。結論:1、高中YKR可有效改善UC大鼠病理狀態(tài),低YKR影響較低。2、高中YKR可以調(diào)整Claudin-2與Claudin-7趨正�；謴�,有效恢復腸道粘膜功能低劑量無效。3、YKR高中劑量組可以有效調(diào)整腸道免疫功能,促使MBL和MASP-2趨于正常狀態(tài)發(fā)展,低劑量無效。
[Abstract]:Aim: to investigate the effects of Yukui recipe (YKR) on the regulation of colonic mucosal barrier function induced by trinitrobenzene sulfonic acid (TNBS) in rats with ulcerative colitis, and to investigate the expression of tight junction protein (Claudin-2) and Claudin-7 and immunoglobulin MBL and MASP-2 in rats with ulcerative colitis. To investigate the function of Yukui recipe in repairing intestinal mucosa and regulating intestinal immune function in rats with ulcerative colitis. Materials and methods: 60 adult SD male rats were randomly assigned into 6 groups. B. C. YKRH dose group, D. YKRI dose group, E. YKRL dose group. Mesalazine group. Except for the normal control group, the rats in the control group were treated with TNBS to induce the onset of UC in SD rats. The rats fasted for 24 hours before the model was made. The distal defecation was stimulated by caressing rats before enema. After intraperitoneal injection of chloral hydrate anesthesia, the rats were suspended upside down. The polyethylene tube with diameter 0.2cm was inserted into the colon 8cm, and the TNBS solution (TNBS was 5% aqueous solution) was infused slowly into the colon, and the TNBS solution was prepared with anhydrous ethanol at 1:1 volume. The final concentration of ethanol was 50%), the dose was 60 mg / kg, the suspension lasted 1 min after enema. On the second day of modeling, it was found that the rats in each group had dilute stool and blood stool, and the rats in group B were given 0.5% sodium carboxymethyl cellulose. After the dose was calculated according to the conversion formula of human body, the rats in group C were given different doses of traditional Chinese medicine by intragastric perfusion. 14 mg / g / d in high dose group, 7 mg / g / d in normal dose group, and intragastric administration of mesalazine suspension of 0.2g/kg/d in low dose 4mg/g/d group and F group, respectively. The rats were given intragastric administration of mesalazine in purified water of 5ml for 14 days, and the rats were killed. The expression of Claudin-2, Claudin-7, MBL and MASP-2 were detected by immunohistochemical technique. Results 1. YKR could effectively improve the symptoms of UC rats. In improving the weight, occult blood and stool of UC rats, YKR high dose group, mesalazine group, middle dose group, YKR low dose group. TNBS induced UC rats to score CMDIDI score after the high dose of YKR. There was no significant difference between the effect and the mesaladine group, and the scores were significantly decreased. However, in the low dose group of YKR, there was no difference between the model group and the model group. 2. TNBS-induced colonic mucosal injury in SD rats, and the increase of Claudin-2 related to tight junction protein decreased Claudin-7, which affected the intestinal mucosal remodeling and permeability. 2. In the middle and high dose group of YKR, the increase of Claudin-7 in Claudin-2 was decreased, and the recovery of intestinal mucosa was improved effectively, but the low dose group was not obvious. MBL protein and MASP-2 protein were decreased in SD rats colon induced by TNBS. MBL protein and MASP-2 protein were increased in middle and high dose group of SD rats induced by TNBS, but not in low dose group. Conclusion: high school YKR can effectively improve the pathological state of UC rats, and the effect of low YKR is low. High school YKR can adjust the normal recovery of Claudin-2 and Claudin-7, and effectively restore intestinal mucosal function. 3YKR high dose group can effectively adjust the intestinal immune function. Promote the development of MBL and MASP-2 in normal state, low dose is not effective.
【學位授予單位】：遼寧中醫(yī)藥大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R285.5

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