發(fā)布時間：2018-01-26 15:02

  本文關(guān)鍵詞： IgAN DCs細胞 T細胞 B細胞　出處：《南昌大學》2016年碩士論文　論文類型：學位論文

【摘要】：目的:通過觀察Ig A腎病(Ig AN)大鼠腸黏膜的免疫細胞數(shù)目和分布的變化從而認識黏膜免疫在Ig AN發(fā)病過程中的作用,比較應(yīng)用大黃酸(RH)前后黏膜免疫細胞的變化探討RH防治Ig AN的作用機理,為RH的臨床應(yīng)用提供理論基礎(chǔ)。方法:將28只SD雌性大鼠隨機分成正常對照組、Ig AN模型組、RH預(yù)防組和RH治療組(n=7)。牛血清蛋白(BSA)+脂多糖(LPS)+四氯化碳(CCl4)聯(lián)合用藥構(gòu)建Ig AN模型,RH預(yù)防組和RH治療組大鼠分別在建模前后用RH懸濁液灌胃。均于10周末殺死大鼠,前一天留取24小時尿液進行尿沉渣紅細胞計數(shù)和24小時尿蛋白的檢測。死后剪部分回腸,進行常規(guī)HE染色,檢測腸黏膜病理變化;取部分回腸和腎組織以液氮保存,OCT包埋,用免疫熒光法檢測腎臟Ig A的沉積,用免疫組織化學法顯示回腸黏膜CD3+T細胞、CD4+T細胞、CD20+B細胞和CD83+樹突狀細胞(DCs細胞)。用方網(wǎng)測試系統(tǒng)測量腸黏膜的固有層DCs細胞和淋巴細胞的數(shù)密度。用統(tǒng)計學軟件SPSS17.0比較各組大鼠固有層樹突狀細胞和淋巴細胞之間的差異和相關(guān)性。結(jié)果:1.Ig AN模型組大鼠尿液檢測可見鏡下血尿,24h尿蛋白含量增加,腎臟免疫熒光染色可見腎皮質(zhì)處有綠色顆粒狀I(lǐng)g A沉積熒光,大黃酸預(yù)防組和大黃酸治療組上述損傷明顯減輕。2.Ig AN模型組大鼠腸道HE染色可見腸絨毛變矮變寬,絨毛間的距離增大,中央乳糜管擴張,固有層結(jié)締組織可見大量炎癥細胞和紅細胞。RH預(yù)防組和RH治療組腸黏膜損傷不同程度改善。3.4組大鼠回腸腸黏膜上皮內(nèi)可見CD3+T細胞。固有層可見CD3+T細胞、CD4+T細胞和CD20+B細胞與CD83+DCs細胞。4.與正常對照組相比,Ig AN大鼠腸黏膜固有層CD3+T細胞、CD4+T細胞、CD20+B細胞和CD83+DCs細胞的數(shù)密度均增加(p0.05)。Ig AN模型組大鼠腸黏膜的固有層CD83+DCs細胞的數(shù)密度與CD4+T細胞的呈顯著性相關(guān)。CD4+T細胞的數(shù)密度與CD20+B細胞的呈高度線性相關(guān)。5.與Ig AN模型組相比,RH預(yù)防組和RH治療組大鼠腸黏膜的固有層CD3+T細胞、CD4+T細胞、CD20+B細胞和CD83+DCs細胞的數(shù)密度均減少(p0.05)。RH預(yù)防組大鼠腸黏膜的固有層CD4+T細胞的數(shù)密度與CD20+B細胞的呈高度線性相關(guān)。RH治療組大鼠腸黏膜的固有層CD4+T細胞的數(shù)密度與CD20+B細胞的呈顯著性相關(guān)。結(jié)論:1.Ig A腎病大鼠腸黏膜固有層CD20+B細胞、CD4+T細胞及CD83+DCs細胞數(shù)密度均增加,提示體液免疫異常活躍,可能與Ig A產(chǎn)生的異常增加和在腎臟的沉積有關(guān)。2 Ig AN大鼠腸黏膜的固有層的DCs細胞與CD20+B細胞及CD4+T細胞與CD20+B細胞之間呈顯著相關(guān)性,提示固有層DCs細胞與淋巴細胞相互作用,共同參與腸道免疫調(diào)節(jié)。3 RH應(yīng)用后能使上述各種異常增高的免疫細胞數(shù)量得到部分恢復,可能是通過抑制DCs細胞的功能和免疫細胞的互相作用來調(diào)節(jié)腸道免疫。
[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.
【學位授予單位】：南昌大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：R277.5

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