IL-17A在口腔黏膜下纤维化中的表达及丹玄口康干预作用研究

发布时间：2018-07-10 09:43

本文选题：IL-17A + 丹玄口康　；参考：《湖南中医药大学》2016年硕士论文

【摘要】：目的:观察IL-17A在大鼠口腔黏膜下纤维化中的表达及丹玄口康干预作用。方法:1、将40只大鼠随机分为A正常组,B模型组,C蒸馏水组,D丹玄口康低剂量组,E丹玄口康高剂量组5组,每组各8只。除A正常组外,其余4组大鼠ANE刺激造模8周。8周后,B组大鼠直接处死并采集口腔颊粘膜组织标本;C、D、E组分别灌胃蒸馏水、低剂量丹玄口康、高剂量丹玄口康4周,处死动物,采集口腔颊粘膜组织标本。观察组织病理学改变,检测胶原沉积状况及IL-17A的表达。2、将30只大鼠随机分为3组,每组10只,分别灌胃蒸馏水、低剂量丹玄口康、高剂量丹玄口康48h后,制备正常血清和含药血清。运用组织块法进行大鼠口腔黏膜成纤维细胞常规培养,取第四代进行实验。将细胞接种后分为4组:A正常组、B ANE组、C丹玄口康低剂量组、D丹玄口康高剂量组,分别加入正常血清、100μg/ml的ANE、丹玄口康低剂量和高剂量血清,干预48h。ELISA检测上清液中羟脯氨酸含量,免疫组化法检测IL-17A的表达。结果:(1)丹玄口康高、低剂量组与蒸馏水组相比,开口度增加,OSF症状有改善;(2)Masson染色结果,模型组与蒸馏水组和正常组相比有明显差异性(P0.01),丹玄口康高、低剂量组与模型组或蒸馏水组相比有显著差异性(P0.01),且高剂量组相对面积更低;(3)免疫组化SABC法测定IL-17A的表达:丹玄口康高、低剂量组IL-17A蛋白表达明显低于模型组或蒸馏水组,存在显著差异(P0.01);(4)用丹玄口康含药血清干预的FB分泌羟脯氨酸明显低于ANE组(P0.01),且丹玄口康高剂量组比丹玄口康低剂量组更低;(5)丹玄口康含药血清干预的FB,可见IL-17A的表达明显低于ANE组,存在显著差异性(P0.01),且丹玄口康高剂量组更明显。结论:IL-17A在实验性OSF大鼠模型中和ANE刺激的大鼠口腔黏膜FB中表达均明显增高,丹玄口康的干预可抑制IL-17A的表达,减少胶原堆积。提示IL-17A可能参与OSF的形成,丹玄口康可以通过抑制IL-17A的表达,抑制胶原堆积,来达到治疗OSF的目的。
[Abstract]:Objective: To observe the expression of IL-17A in the Oral Submucosal Fibrosis in rats and the intervention of Dan Xuan Kou Kang. Methods: 1, 40 rats were randomly divided into A normal group, B model group, C distilled water group, D Dan Xuan Kou Kang low dose group, E Dan Xuan Kou Kang high dose group 5 groups, each group except the A normal group, the other 4 groups of rats ANE stimulation after 8 weeks.8 weeks, B group, B group Rats were executed directly and collected buccal mucosa tissue specimens. C, D and E group were treated with distilled water, low dose Dan Xuan Kou Kang, and high dose of Dan Xuan Kou Kang for 4 weeks. The animals were killed and the buccal mucosa tissue specimens were collected. The histopathological changes were observed and the collagen deposition and the expression of IL-17A were detected. The 30 rats were randomly divided into 3 groups, 10 rats in each group, divided into 3 groups. The normal serum and drug containing serum were prepared after 48h, with low dose of Dan Xuan Kou Kang and high dose of Dan Xuan Kou Kang. The normal culture of rat oral mucosa fibroblasts was carried out by tissue block method, and the experiment was carried out for fourth generations. The cells were divided into 4 groups: normal A group, B ANE group, C Dan Xuan Kou Kang low dose group, D Dan Xuan Kou Kang high dose group, Adding normal serum, 100 g/ml ANE, Dan Xuan Kou Kang low dose and high dose serum, intervention 48h.ELISA to detect the content of hydroxyproline in the supernatant, immunohistochemical method to detect the expression of IL-17A. Results: (1) Dan Xuan Kou Kang Gao, lower dose group compared with the distilled water group, the opening degree increased, the OSF symptoms improved; (2) Masson staining results, model group and Compared with the normal group, the distilled water group had significant difference (P0.01), the lower dose group was significantly different from the model group or the distilled water group (P0.01), and the high dose group was lower in relative area. (3) the expression of IL-17A by immunohistochemical SABC method: Dan Xuan Kou Kang Gao, the low dose group IL-17A protein expression was obviously lower than the model group or distillation. There were significant differences (P0.01) in the water group (P0.01); (4) the FB secreted hydroxyproline in Dan Xuan Kou Kang serum was significantly lower than that of the ANE group (P0.01), and the high dose group of Dan Xuan Kou Kang dose group was lower than that of the low dose group of Dan Xuan Kou Kang; (5) the FB of the serum intervention of Dan Xuan Kou Kang containing medicine showed that the expression of IL-17A was significantly lower than that of the ANE group, and there was a significant difference (P0.01), and Dan Xuan Kou was found. Conclusion: the expression of IL-17A in the experimental OSF rat model and the ANE stimulated rat oral mucosa FB were significantly increased. The intervention of Dan Xuan Kou Kang can inhibit the expression of IL-17A and reduce the accumulation of collagen. It suggests that IL-17A may be involved in the formation of OSF, and Dan Xuan Kou Kang can inhibit the accumulation of collagen by inhibiting the expression of IL-17A, and inhibiting the accumulation of collagen. To achieve the purpose of treating OSF.
【学位授予单位】：湖南中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R276.8

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