正常腎小管細胞質8-OHdG免疫組織化學染色表現與腎臟纖維化相關

Intensity of Cytosol Expression of 8-OHdG in Normal Renal Tubules Is Sssociated with The Severity of Renal Fibrosis

劉嘉鴻(Jia-Hung Liou)

指導教授 ： 楊順發

中山醫學大學/醫學院/醫學研究所/碩士(2013年)

https://doi.org/10.6834/CSMU.2013.00227

摘要

慢性腎臟病（又稱慢性腎衰竭），係腎功能在幾個月到數年期間逐漸而難以逆轉的衰退。根據中華國民衛生署統計，腎炎、腎病症候群及腎病變，在2011年名列國人十大死因的第十位，而大多數患者對自身病情往往並不自知。慢性腎臟病的主要病理檢查下的組織特徵就是腎臟纖維化。目前已知活性氧於腎臟纖維化過程中扮演著重要角色。本研究擬找出適合於病理檢查下除了纖維化程度之觀察外，另一個可以預期腎功能狀況的預後評估因子。8-OHdG是敏感的DNA損害標誌物，由一個氫氧基接在鳥嘌呤的第8個碳上而形成，8-OHdG是白血球受到氧化傷害之後的代謝物，目前8-OHdG已被當成細胞內DNA氧化壓力的重要指標。本研究利用74例腎臟切除檢體使用免疫組織化學染色，去分析8-OHdG在腎臟組織內各個成分組成的表現量與腎臟纖維化程度及各項臨床病理特徵的相關性。
結果發現，腎間質纖維化分數與腎絲纖維化分數高的病例，同時也會具有較高的血清肌酐酸濃度，較低的腎絲球過濾率，較高比例罹患慢性腎臟病或泌尿上皮癌。而在分析腎間質纖維化分數較高的病例群時，也發現會有較低的正常腎小管的細胞質8-OHdG染色濃度(35.7% vs. 64.3%, p =0.011)，及較低的正常腎小管的細胞核8-OHdG染色濃度(28.6% vs. 71.4%,p = 0.023)。單變項分析發現腎間質纖維化分數與腎絲球纖維化分數皆和正常腎小管的細胞質8-OHdG染色濃度呈現正相關，但腎間質纖維化分數卻是與正常腎小管的細胞核8-OHdG染色濃度呈現負相關。再進一步以多變項分析後，發現了血清肌酐酸濃，及正常腎小管的細胞質8-OHdG染色濃度為獨立的腎間質纖維化分數與腎絲纖維化分數預估因子。根據本研究結果認為，正常腎小管的細胞質8-OHdG染色濃度可以做為腎臟纖維化的預後評估因子。

關鍵字

慢性腎臟病 ； 8-羥基去氧鳥糞嘌呤核 ； 免疫組織化學染色 ； 腎臟纖維化 ； 活性氧化物

並列摘要

Chronic renal disease (also known as chronic renal failure) means irreversible decline of renal function for months or years. According to the records from Department of health, R.O.C (Taiwan), renal disease (includes nephritis, nephrotic syndrome, and nephropathy) was the tenth cause of death in Taiwan in 2011. The most characteristic pathologic finding of chronic renal disease was renal fibrosis. It had been proposed that reactive oxygen species play a role in renal fibrosis. 8-OHdG, a metabolite of oxidative damage to leukocyte DNA, has been identified as a marker of oxidative stress in patients with chronic renal failure.
Seventy-four patients following nephrectomy were retrospectively enrolled. Immunohistochemical analysis of the renal expression of 8-OHdG in the nephrectomised kidneys was performed and associations between renal expression of 8-OHdG and renal fibrosis were evaluated.
Patients with higher interstitial fibrosis scores (IFS) and glomerular fibrosis scores (GFS) had significantly higher serum creatinine, lower estimated glomerular filtration rate (eGFR), increased percentage of chronic kidney disease (CKD) and urothelial cell carcinoma. The renal tissues with higher IFS had lower expressions of 8-OhdG in normal tubular cytoplasm (NTc) (35.7% vs. 64.3%, p = 0.011) and normal tubular nuclei (NTn) (28.6% vs. 71.4%, p = 0.023). Univariate analysis showed that IFS and GFS correlated with the NTc 8-OHdG expression and IFS negatively correlated with NTn 8-OHdG expression. Multivariate stepwise regression revealed that serum creatinine (r = 0.351 for IFS, p = 0.021; r = 0.563 for GFS, p <0.001) and intensity of 8-OHdG expression in NTc (r = 0.397 for IFS, p = 0.01; r = 0.278 for GFS, p = 0.043) were the independent factors predicting IFS or GFS. This study demonstrated that the intensity of 8-OHdG expression in NTc was associated with the severity of renal fibrosis.

並列關鍵字

chronic kidney disease ； immunohistochemical stain ； renal fibrosis ； reactive oxygen species

參考文獻

Akcay T, Saygili, Angican G, et al. Increased formation of 8-hydroxy-2'-deoxyguanosinein peripheral blood leukocytes in bladder cancer. Urol Int, 71(3):271–4, 2003.

Brenner BM. Remission of renal disease: recounting the challenge, acquiring the goal. J Clin Invest, 110(12):1753–8, 2002.

Cadenas E, Davies KJ. Mitochondrial free radical generation, oxidative stress, and aging. Free Radic Biol Med, 29(3-4):222-30, 2000.

Chang CH, Yang CM, Yang AH. Renal diagnosis of chronic hemodialysis patients with urinary tract transitional cell carcinoma in Taiwan. Cancer, 109(8):1487–92, 2007.

Chen CH, Dickman KG, Moriya M, et al. Aristolochic acid-associated urothelial cancer in Taiwan. Proc. Natl. Acad. Sci. U.S.A., 109 (21): 8241–6, 2012.