【摘要】：目的:(1)應(yīng)用血清蛋白質(zhì)組學(xué)探討糖尿病腎病(Diabetic Nephropathy,DN)發(fā)生發(fā)展機制。(2)驗證甘露糖結(jié)合凝集素相關(guān)絲氨酸蛋白酶2(MBL associated serine protease 2,MASP2)在糖尿病腎病不同階段的表達水平。(3)探討甘露聚糖結(jié)合凝集素(mannose binding lectin,MBL)與2型糖尿病腎病的關(guān)系。方法:第一部分:入選年齡、性別匹配研究對象共4組,每組8人。組A為正常對照組,組B、C、D分別為2型糖尿病尿微量白蛋白排泄率(UAER)㩳30mg/g肌酐組、30mg/g肌酐≤UAER㩳300mg/g肌酐組、UAER≥300mg/g肌酐組。利用雙向電泳對4組血清進行蛋白質(zhì)分離。應(yīng)用Image Master 2D Platinum分析軟件(GE公司)分析四組之間差異表達的蛋白質(zhì)點。挖取表達差異蛋白質(zhì)點,進行基質(zhì)輔助激光解附離子化飛行時間質(zhì)譜(MALDI-TOF MS)分析,并與Swiss Prot數(shù)據(jù)庫進行匹配鑒定蛋白質(zhì)。第二部分:入選正常對照組(A組)54人,2型糖尿病患者尿微量白蛋白30mg/g肌酐組(B組)52人、尿微量白蛋白≥30mg/g肌酐且300mg/g肌酐組(C組)30人、尿微量白蛋白≥300mg/g肌酐組(D組)16人。采用雙抗體夾心酶聯(lián)免疫吸附實驗(ELISA)來檢測4組血清MASP2水平,驗證血清蛋白質(zhì)組實驗結(jié)果。同時應(yīng)用ELISA方法檢測以上四組血清MBL水平。并分析MASP2和MBL與2型糖尿病慢性并發(fā)癥的關(guān)系。結(jié)果:第一部分:血清蛋白質(zhì)組學(xué)結(jié)果顯示,在2型糖尿病B組患者中MASP2表達上調(diào)。第二部分:ELISA實驗結(jié)果顯示:(1)2型糖尿病組MASP2水平高于正常對照組(P0.05);(2)2型糖尿病B組B組和D組的MASP2表達高于C組(P0.05);(3)MBL在正常對照組與糖尿病組沒有差別(P0.05),不同糖尿病腎病組沒有差別(P0.05)。結(jié)論:(1)2型糖尿病患者甘露糖結(jié)合凝集素相關(guān)絲氨酸蛋白酶2水平升高。(2)MASP2可能參與2型糖尿病腎病的發(fā)生發(fā)展.
[Abstract]:Objective: (1) to investigate the mechanism of diabetic nephropathy (Diabetic Nephropathy,DN) by serum proteome. (2) to verify the expression of mannose binding lectin associated serine protease 2 (MBL associated serine protease 2, MASP2) in different stages of diabetic nephropathy. (3) to explore the relationship between mannan binding lectin (mannose binding lectin,MBL and type 2 diabetic nephropathy. Methods: in the first part, there were 4 groups of age and sex matching, 8 people in each group. Group A was normal control group, group B, C, D were type 2 diabetes mellitus urinary albumin excretion rate (UAER) 30mg/g creatine group, 30mg/g creatine 鈮，

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