Title: 骨橋蛋白以及小分子核醣核酸196a在糖腎病的影響
The Effect of Osteopontin (OPN) and miR196a in Diabetic Nephropathy (DN)
Authors: 白喬之
Pai, Chiao-Chih
Wang, Chih-Hong
Keywords: 糖腎病;骨橋蛋白;小分子核醣核酸196a;Diabetic nephropathy;osteopontin;miR196a
Issue Date: 2017
Abstract: 近代社會中,隨著速食、高糖的飲食習慣增加以及缺少規律運動的生活作息,第二型糖尿病的患病率逐年成長。臨床數據顯示,許多第二型糖尿病患者會惡化為末期腎臟疾病或者稱為腎衰竭或腎臟纖維化,並需要依靠腎臟移植或是洗腎來維持生命。近年實驗研究中發現,小分子核糖核酸196a透過增加動物體內棕色脂肪組織生成以調控能量利用,進而有助於肥胖患者調節體重的醫療潛能。然而在miR196a-Hoxc8路徑下游蛋白中的骨橋蛋白(Osteopontin) 是在糖尿病患者血清中發現有高表現量且在近期研究中被視為造成纖維化的危險因子之一。因此我們的計劃專注於找到miR196a以及OPN在第二型糖尿病中造成的影響 實驗中,我們利用第二型糖尿病動物模型Leprdb/db小鼠與miR196a基因轉殖小鼠配種,來取得全身性大量表現miR196a的糖尿病小鼠(miR196a; Leprdb/db mice)以進行實驗。藉由qPCR mRNA的測量,我們發現在miR196a表現的糖尿病小鼠中,OPN的表現量有顯著下降且其腎臟中的發炎因子IL-1B、 iNOS、F4/80也呈現明顯的減低趨勢。另外,在西方墨點法蛋白質測量中也顯示了miR196a糖尿病小鼠腎臟中的間質相互轉換(EMT)蛋白Vimentin比普通糖尿病鼠低許多。上述的實驗結果初步的指出,在第二型糖尿病腎臟中miR196a有調控OPN表限量減低並降低發炎因子表現進而減輕腎臟受損的潛能。未來,這份計畫會進一步增加腎臟組織切片以更直接明確的觀察腎臟部位的受損狀況以及纖維化程度,並以了解更細節的細胞路徑調控為目標前進。
With a lifestyle characterized by low physical activity and high-energy food intake, the prevalence of Type 2 Diabetic Mellitus(T2DM) increased dramatically these years. Clinically, many T2DM patients end in End Stage Renal Disease(ESRD) and need to undergo dialysis to prolong their lives. In research, miR196 was reported to be a potential therapeutic target for inducing brown adipogenesis to combat T2DM. However, the miR196-Hoxc8 pathway has a potential effect of regulating the osteopontin (OPN) which is reported to have a high level in the T2DM patients’ serum. The aim of our research is focused on the effect of OPN and miR196a in T2DM. For this purpose, we used Leprdb/db mice for the T2DM animal model and cross-breeding with miR196a transgenic mice to get the miR196a; Leprdb/db mice for our experiment. The metabolic cage measurement and mice tissue were collected for in vivo experiment. For the real-time qPCR results, we found the OPN level decrease and less inflammation factor IL-1B, iNOS and F4/80 expression in miR196a group compared to the normal T2DM group. Also, in western blot experiment, we found that both EMT marker Vimentin decrease in miR196a group. The above results showed that miR196 have the potential function of down-regulating the OPN expression and protect the DM kidney from diabetic nephropathy. In the future direction, we will try to find the detail pathway of miR196a to OPN and finish the histology to compare the fibrosis level in the two group.
URI: http://etd.lib.nctu.edu.tw/cdrfb3/record/nctu/#GT070457011
Appears in Collections:Thesis