文献类型：学位论文

中文题名：温肾健脾法对溃疡性结肠炎模型大鼠血清IL-8、结肠组织NF-κBp65、IL-1βmRNA表达影响的研究

英文题名：The Research of Warming Kidney and Invigorating Spleen Influences on Expression of Serum IL-8, Colon Tissue NF-κBp65, IL-1β mRNA of Ulcerative Colitis Model Rats

作者：荀敏奇[1];

第一作者：荀敏奇

机构：[1]甘肃中医学院;

第一机构：甘肃中医药大学

导师：李兰珍;甘肃中医学院

授予学位：硕士

语种：中文

中文关键词：UC;温肾健脾法;脾肾阳虚型;IL-8;NF-κBp65;IL-1β

外文关键词：ulcerative colitis;warming kidney and invigorating spleen method;the spleen kidney Yang deficiency syndrome;IL-8;NF-κBp65;IL-1β

年份：2014

摘要：目的 1、阐明脾肾阳虚证型是UC的重要证型，温肾健脾法及其代表方四神丸是治疗复发型、难治型UC的重要治法、方药。 2、初步阐释温肾健脾法及其代表方四神丸调控UC模型大鼠肠黏膜修复的作用机制，为用温肾健脾法治疗UC提供科学依据。 方法 采用“病—证结合”的方法（番泻叶浸液灌胃+糖皮质激素腹腔注射+TNBS/乙醇混悬液灌肠）制备脾肾阳虚证UC模型大鼠。之后采用高、中、低三种不同剂量的四神丸进行干预治疗21天，同时以SASP作为阳性对照药物进行治疗。取材后，采用病理学方法（肉眼观察结肠组织大体形态并评分、HE染色镜下观察肠黏膜损伤及修复情况）观察大鼠结肠组织修复情况、ELISA法检测大鼠血清IL-8、SP法和qRT-PCR法分别检测大鼠结肠组织中NF-κBp65、IL-1β的表达。 结果 1、ELISA法检测大鼠血清IL-8表达：与空白组比较，模型组大鼠血清IL-8含量显著增高（P<0.01）；治疗后，与模型组比较，对照组、四神丸低、中、高剂量组大鼠血清IL-8含量均显著降低（P<0.01）。 2、SP法检测大鼠结肠组织NF-κBp65表达：（1）镜下可见：空白组NF-κBp65阳性细胞呈阴性表达，模型组NF-κB65阳性细胞呈胞浆阳性，阳性细胞为单核-巨噬细胞，散在浸润于黏膜下层、淋巴滤泡内及外膜层，呈中等强度至强阳性表达。其余各治疗组大鼠NF-κB65阳性细胞均成呈弱度至中等强度表达。（2）NF-κB65阳性细胞平均灰度值结果显示：与空白组比较，模型组NF-κB65表达显著增强（P<0.01）；治疗后，与模型组比较，对照组、四神丸中、高剂量组NF-κB65表达显著减弱（P<0.01）。 3、RT-qPCR法检测大鼠结肠组织IL-1β mRNA表达：与空白组比较，模型组IL-1βmRNA表达显著增强（P<0.01）；治疗后，与模型组比较，对照组、四神丸中、高剂量组IL-1β mRNA表达显著减弱（P<0.05）。 结论 1、四神丸降低了UC模型大鼠血清中IL-8的表达，抑制了导致机体炎症反应的活性产物的释放，从而减弱了机体炎症反应。 2、四神丸降低了UC模型大鼠结肠组织中NF-κBp65的表达，终止了相关炎症反应基因的转录，使炎症免疫细胞增殖及细胞凋亡减少，从而起到抑制炎症免疫反应的作用。 3、四神丸降低了UC模型大鼠结肠组织中IL-1β的表达，阻碍了中性粒细胞向病变结肠部位的募集和趋化，减少了肠黏膜的损伤，从而有效地促进大鼠肠黏膜的修复，最终达到治疗疾病的作用。
Objective 1. Clarify the spleen kidney Yang deficiency syndrome is the most important syndrome inUC, the method of warming kidney and invigorating spleen is important treatment, and it’son behalf of the Sishen Wan is important Chinese herbal medicine on curing recurrent andrefractory UC. 2. Preliminary interpret the mechanism of UC model rats’ intestinal mucosa repair about themethod of warming kidney and invigorating spleen and it’s on behalf of the Sishen Wan.Provide a scientific basis for the method of warming kidney and invigorating spleen curesUC. Method Adopt the method of “combined disease-and-syndrome”/(Fenxieye gavage+glucocorticoid intraperitoneal injection+TNBS//ethanol mixed suspension clyster/) preparingspleen kidney Yang deficiency syndrome UC rats model. After the high, medium and lowthree different doses of the Sishen Wan to intervene treatment for21days, at the same timewith SASP treatment as a positive control drug. Then put all the rats to death and dissect.Observe the colon tissue pathological morphology of rats, and detect the serum IL-8level ofrats with ELISA, and NF-κBp65with SP, IL-1β with qRT-PCR expression. Result 1. ELISA method detects the rats serum IL-8expression: compared with the blank group,model group rats serum IL-8level increased significantly /(P <0.01/); after treatment,compared with model group, control group, Sishenwan low, medium and high dose grouprats serum IL-8level reduced significantly /(P <0.01/). 2. SP method detects the rat colonic tissue NF-κBp65expression:/(1/) microscopically: theblank group NF-κB65positive cells are negative expression, and the model group NF-κB65positive cells which are monocytes infiltrated scatteredly in the submucosa, lymphoidfollicles and outer membrane layer are positive expression in cytoplasm, appear moderate orhigher intensity expression. The rest of the treatment group rats NF-κB65positive cellsappear lower or moderate intensity expression./(2/) The NF-κB65positive cells average grey value results show that compared with the blank group, the NF-κB65expression of modelgroup significantly enhanced /(P<0.01/); after treatment, compared with model group, theNF-κB65expression of control group, Sishenwan medium and high dose groups significantlyreduced /(P<0.01/). 3. RT-qPCR method detects the expression of rat colonic tissue IL-1β mRNA: compared withblank group, the expression of model group IL-1β mRNA significantly enhanced /(P<0.01/);after treatment, compared with model group, the expression of control group, Sishenwanmedium and high dose group IL-1β mRNAsignificantly reduced /(P<0.05/). Conclusion 1. Sishenwan reduces the UC model rats serum IL-8expression, and suppresses theinflammatory response in the body of the release of active products, thus weakens the body'sinflammatory response. 2. Sishenwan reduces the UC model rats colon tissue NF-κBp65expression, and terminatesthe related inflammation gene transcription, and reduces inflammation immune cellproliferation and apoptosis to have the effect of inhibiting the inflammatory reaction. 3. Sishenwan reduces the UC model rats colon tissue of the expression of IL-1β, andhindereds the raise of neutrophils to colon and reduces the intestinal mucosal damage.Effectively promote restoration of the intestinal mucosa of rats. Finally, reach the role oftreating diseases.