發(fā)布時間：2018-06-16 18:49

  本文選題：IgA腎病 + 腎小管間質纖維化　； 參考：《安徽醫(yī)科大學》2016年碩士論文

【摘要】：目的：探討巨噬細胞在腎小管間質纖維化(Tubulointerstitial fibrosis, TIF)及上皮細胞間充質轉化(Epithelial-Mesenchymal Transition, EMT)中的作用及可能機制,為臨床上腎臟疾病針對TIF的治療提供理論依據。方法：(1)收集安徽醫(yī)科大學第二附屬醫(yī)院病理科56例腎臟穿刺活檢診斷為IgA腎病患者標本,用HE和特殊染色PAS、PASM、Masson染色法檢測腎間質病理變化,根據腎間質纖維化及腎小管萎縮程度將56例病例分為輕、中、重度三組,另收集5例腎腫瘤遠離腫瘤的正常腎組織為實驗對照組。采用免疫組化方法,用CD68標記巨噬細胞,CD206標記M2型巨噬細胞,同時檢測EMT相關蛋白TGF-β1、Vimentin、SMA、E-Cadherin的表達,并收集患者詳細臨床資料：(2)人腎小管上皮細胞HK-2與不同濃度巨噬細胞U937共培養(yǎng)。將實驗分為五組：空白對照組(HK-2細胞單獨培養(yǎng)),實驗1組(HK-2細胞與1×104個/ml濃度的U937細胞共培養(yǎng)),實驗2組(HK-2細胞與1×105個/ml濃度的U937細胞共培養(yǎng)),實驗3組(HK-2細胞與1×106個/ml濃度的U937細胞共培養(yǎng)),實驗4組(HK-2細胞與1×107個/ml濃度的U937細胞共培養(yǎng))。倒置顯微鏡下觀察細胞形態(tài)變化,采用Western blot方法檢測EMT相關蛋白α-SMA、 Vimentin及E-cadherin的表達。結果：(1) CD68、CD206陽性細胞在TIF重度組高于中度組,在TIF中度組高于TIF輕度組(P0.05)；CD68陽性細胞、CD206陽性細胞與TGF-β1、SMA、Vimentin蛋白的表達呈正相關(P0.05)；與E-Cadherin蛋白的表達負相關(P0.05)；與腎功能指標血肌酐、尿素氮及尿蛋白正相關。TGF-β1、SMA、Vimentin蛋白的表達與腎小管間質纖維化程度及腎功能指標血肌酐、尿素氮及尿蛋白正相關(P0.05);E-Cadherin的表達與腎小管間質纖維化程度及上述腎功能指標呈負相關(P0.05)。(2)與U937細胞共培養(yǎng)24h后,HK-2細胞形態(tài)由鋪路石樣變?yōu)殚L梭形,細胞間隙明顯增寬；與對照組比較,與U937細胞共培養(yǎng)后HK-2細胞α-SMA、Vimentin蛋白的表達明顯增加,E-cadherin蛋白的表達明顯下降,差異具有統計學意義(P0.05)。結論：巨噬細胞在腎臟疾病中可能具有促進腎間質纖維化及腎小管上皮細胞EMT的作用。
[Abstract]:Objective: to investigate the role and possible mechanism of macrophages in tubulointerstitial fibrosis (TIFs) and epithelial mesenchymal transition (EMTT). Methods the specimens of 56 patients with IgA nephropathy diagnosed by renal biopsy were collected from Department of Pathology, second affiliated Hospital of Anhui Medical University. The pathological changes of renal interstitium were detected by HE and PAS-PASMMA-Masson staining. According to the degree of interstitial fibrosis and atrophy of renal tubules, 56 cases were divided into three groups: mild, moderate and severe. M 2 macrophages were labeled with CD68 and CD206, and the expression of EMT associated protein TGF- 尾 1 and VimentinnSMAE-Cadherin was detected by immunohistochemistry. The detailed clinical data of human renal tubular epithelial cells (HK-2) were collected and co-cultured with different concentrations of macrophages (U937). The experiment was divided into five groups: the blank control group was cultured alone, the experimental group 1 was co-cultured with 1 脳 104 / ml U937 cells, the second group was co-cultured with 1 脳 105 / ml U937 cells, and the third group was treated with U937 cells at a concentration of 1 脳 105 / ml. The cells were co-cultured with 1 脳 106 / ml U937 cells and 1 脳 107 / ml U937 cells. The expression of EMT related proteins 偽 -SMA, vimentin and E-cadherin were detected by Western blot. Results (1) the positive cells of CD68 and CD206 were positively correlated with the expression of TGF- 尾 1 SMAMA-Vimentin protein and negatively correlated with the expression of E-Cadherin protein in TIF severe group, and in TIF moderate higher than that in TIF mild TIF group (P 0.05), and the expression of TGF- 尾 _ 1SMAMA-Vimentin protein was negatively correlated with the expression of E-Cadherin protein (P0.05A) in TIF group and TGF- 尾 _ 1 SMAMA-Vimentin protein. The expression of Vimentin protein was positively correlated with serum creatinine, urea nitrogen and urine protein. The expression of Vimentin protein was correlated with the degree of renal tubulointerstitial fibrosis and the serum creatinine of renal function index. The expression of E-Cadherin in U937 cells and U937 cells were negatively correlated with the degree of renal tubulointerstitial fibrosis and the above renal function indexes. After 24 hours of co-culture with U937 cells, the morphology of HK-2 cells changed from paving stone to long fusiform, and the intercellular gap was obviously widened. Compared with the control group, the expression of 偽 -SMA-Vimentin protein in HK-2 cells increased significantly after co-culture with U937 cells, and the expression of E-cadherin protein decreased significantly (P 0.05). Conclusion: macrophages may play a role in promoting renal interstitial fibrosis and EMT in renal tubular epithelial cells.
【學位授予單位】：安徽醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：R692

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