IgA肾病大鼠肠黏膜免疫细胞的变化及大黄酸的调节作用

发布时间：2018-01-26 15:02

本文关键词： IgAN DCs细胞 T细胞 B细胞　出处：《南昌大学》2016年硕士论文　论文类型：学位论文

【摘要】：目的:通过观察Ig A肾病(Ig AN)大鼠肠黏膜的免疫细胞数目和分布的变化从而认识黏膜免疫在Ig AN发病过程中的作用,比较应用大黄酸(RH)前后黏膜免疫细胞的变化探讨RH防治Ig AN的作用机理,为RH的临床应用提供理论基础。方法:将28只SD雌性大鼠随机分成正常对照组、Ig AN模型组、RH预防组和RH治疗组(n=7)。牛血清蛋白(BSA)+脂多糖(LPS)+四氯化碳(CCl4)联合用药构建Ig AN模型,RH预防组和RH治疗组大鼠分别在建模前后用RH悬浊液灌胃。均于10周末杀死大鼠,前一天留取24小时尿液进行尿沉渣红细胞计数和24小时尿蛋白的检测。死后剪部分回肠,进行常规HE染色,检测肠黏膜病理变化;取部分回肠和肾组织以液氮保存,OCT包埋,用免疫荧光法检测肾脏Ig A的沉积,用免疫组织化学法显示回肠黏膜CD3+T细胞、CD4+T细胞、CD20+B细胞和CD83+树突状细胞(DCs细胞)。用方网测试系统测量肠黏膜的固有层DCs细胞和淋巴细胞的数密度。用统计学软件SPSS17.0比较各组大鼠固有层树突状细胞和淋巴细胞之间的差异和相关性。结果:1.Ig AN模型组大鼠尿液检测可见镜下血尿,24h尿蛋白含量增加,肾脏免疫荧光染色可见肾皮质处有绿色颗粒状Ig A沉积荧光,大黄酸预防组和大黄酸治疗组上述损伤明显减轻。2.Ig AN模型组大鼠肠道HE染色可见肠绒毛变矮变宽,绒毛间的距离增大,中央乳糜管扩张,固有层结缔组织可见大量炎症细胞和红细胞。RH预防组和RH治疗组肠黏膜损伤不同程度改善。3.4组大鼠回肠肠黏膜上皮内可见CD3+T细胞。固有层可见CD3+T细胞、CD4+T细胞和CD20+B细胞与CD83+DCs细胞。4.与正常对照组相比,Ig AN大鼠肠黏膜固有层CD3+T细胞、CD4+T细胞、CD20+B细胞和CD83+DCs细胞的数密度均增加(p0.05)。Ig AN模型组大鼠肠黏膜的固有层CD83+DCs细胞的数密度与CD4+T细胞的呈显著性相关。CD4+T细胞的数密度与CD20+B细胞的呈高度线性相关。5.与Ig AN模型组相比,RH预防组和RH治疗组大鼠肠黏膜的固有层CD3+T细胞、CD4+T细胞、CD20+B细胞和CD83+DCs细胞的数密度均减少(p0.05)。RH预防组大鼠肠黏膜的固有层CD4+T细胞的数密度与CD20+B细胞的呈高度线性相关。RH治疗组大鼠肠黏膜的固有层CD4+T细胞的数密度与CD20+B细胞的呈显著性相关。结论:1.Ig A肾病大鼠肠黏膜固有层CD20+B细胞、CD4+T细胞及CD83+DCs细胞数密度均增加,提示体液免疫异常活跃,可能与Ig A产生的异常增加和在肾脏的沉积有关。2 Ig AN大鼠肠黏膜的固有层的DCs细胞与CD20+B细胞及CD4+T细胞与CD20+B细胞之间呈显著相关性,提示固有层DCs细胞与淋巴细胞相互作用,共同参与肠道免疫调节。3 RH应用后能使上述各种异常增高的免疫细胞数量得到部分恢复,可能是通过抑制DCs细胞的功能和免疫细胞的互相作用来调节肠道免疫。
[Abstract]:Objective: to investigate the role of mucosal immunity in the pathogenesis of IgAN by observing the changes of the number and distribution of immune cells in the intestinal mucosa of rats with IgA nephropathy. Objective: to compare the changes of mucosal immune cells before and after the application of Rhein (RH) to explore the mechanism of RH in the prevention and treatment of IgAN. Methods: 28 SD female rats were randomly divided into normal control group and IgAN model group. The IgAN model was established by combined use of RH prevention group and RH treatment group (BSA) lipopolysaccharide (LPSL) CCL 4). Rats in RH prevention group and RH treatment group were fed with RH suspension before and after modeling and killed at the end of 10 weeks. Urine samples were collected 24 hours before the day before, and urine sediment erythrocyte count and 24 hour urine protein were detected. After death, part of ileum was cut and routine HE staining was used to detect the pathological changes of intestinal mucosa. Some of the ileum and kidney tissues were preserved with liquid nitrogen and Oct. The deposition of IgA in kidney was detected by immunofluorescence method. The CD4 T cells of CD3 T cells in ileal mucosa were detected by immunohistochemical method. CD20 B cells and CD83 dendritic cells. The number density of DCs cells and lymphocytes in the lamina propria of intestinal mucosa were measured by square net test system. The difference between dendritic cells and lymphocytes in lamina propria of rats was compared by statistical software SPSS17.0. Results:. 1.The urine of IgAN model group was detected by hematuria under microscope. 24 hours urine protein content increased, kidney immunofluorescence staining showed green granular Ig A deposition fluorescence in the renal cortex. Rhein prevention group and Rhein treatment group reduced the above injury significantly. 2. HE staining showed that intestinal villi became shorter and wider, the distance between villi increased, and the central chylotube dilated in IgAN model group. In the connective tissue of lamina propria, a large number of inflammatory cells and red blood cells. RH prevention group and RH treatment group showed different degree of improvement of intestinal mucosal injury. 3. 4 rats showed CD3 in ileal mucosal epithelium. CD3 T cells were found in the lamina propria. CD4 T cells and CD20 B cells and CD83 DCs cells. 4. Compared with the normal control group, CD3 T cells in the lamina propria of IgAN rats had CD4 T cells. The number density of CD20 B cells and CD83 DCs cells increased by p0.05). The number density of CD83 DCs cells in the lamina propria of the intestinal mucosa in the IgAN model group was significantly correlated with the number density of CD4 T cells. The number density of CD4 T cells was significantly correlated with the number density of CD20. B cells showed a high linear correlation. 5. Compared with the IgAN model group. The CD3 T cells in the lamina propria of rats in RH prevention group and RH treatment group were CD4 T cells. The number density of CD20 B cells and CD83 DCs cells decreased by p0.05). The number density of CD4 T cells in the lamina propria of rat intestinal mucosa in RH preventive group was highly linearly correlated with that of CD20 B. CD4 in the lamina propria of rat intestinal mucosa in RH treatment group was highly linear. The number density of T cells was significantly correlated with the number of CD20 B cells. Conclusion 1. CD20 B cells in the lamina propria of intestinal mucosa of rats with IgA nephropathy. The number density of CD4 T cells and CD83 DCs cells increased, suggesting that humoral immunity was abnormal. DCs cells and CD20 B cells and CD4 T cells and CD20 in the lamina propria of intestinal mucosa in IGAN rats may be associated with abnormal increase in IgA production and deposition in the kidney. There was a significant correlation between B cells. These results suggest that the interaction between DCs cells and lymphocytes in lamina propria can partially restore the number of these abnormal immune cells after the use of intestinal immunomodulation 3. 3 RH. Intestinal immunity may be regulated by inhibiting the function of DCs cells and the interaction of immune cells.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R277.5

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