MiR-29b在紫癜性腎炎腎組織中的表達及其功能研究
發(fā)布時間:2018-05-20 01:27
本文選題:紫癜性腎炎 + 新月體 ; 參考:《南京醫(yī)科大學》2013年碩士論文
【摘要】:第一部分MiR-29b在紫癜性腎炎腎組織中的表達 目的檢測miR-29b在兒童紫癜性腎炎(Henoch Sch nlein purpura nephritis,HSPN)腎組織中的表達,并探討其與HSPN病理損傷的關(guān)系。 方法35例HSPN患兒活檢腎組織,光鏡和電鏡下觀察各患兒的病理變化,按病理變化有無新月體形成、腎小球硬化、間質(zhì)纖維化、血栓形成、毛細血管袢壞死各分為兩組。采用real-time PCR的方法定量檢測腎活檢組織中miR-29b的表達水平,并記錄患兒入院后首次檢查的實驗室指標,用SPSS17.0軟件分析兩組間miR-29b表達水平及實驗室指標的差異,進一步聯(lián)合miR-29b及實驗室指標建立Logistic回歸模型,分析其與HSPN新月體形成的關(guān)系。 結(jié)果(1)與無新月體組相比,有新月體組miR-29b表達水平增加(P0.05),其他病理改變組miR-29b表達水平無變化。(2)無新月體組和有新月體組的入院后實驗室指標比較中,血小板計數(shù)(PLT)、中性粒細胞百分比(N%)、嗜酸性粒細胞百分比(E%)、尿白細胞計數(shù)(UWBC)、尿紅細胞計數(shù)(URBC)、體重校正后24h尿蛋白定量(24hUPro/kg)、血清膽固醇(CHOL)及血IgG水平在兩組間有統(tǒng)計學差異(P0.05)。(3)聯(lián)合miR-29b與有統(tǒng)計學差異的實驗室指標進行Logistic回歸分析,結(jié)果發(fā)現(xiàn)miR-29b(OR=2.166,P=0.041),24hUPro/kg(OR=5.296,P=0.042),URBC(OR=6.016,P=0.033)進入Logistic回歸模型(LogP=-4.633+0.773miR-29b+1.66724hUPro/kg+1.794URBC),作為預測新月體形成的獨立危險因素。 結(jié)論伴新月體形成的HSPN腎組織中miR-29b表達水平增加。miR-29b、24hUPro/kg及URBC是紫癜性腎炎新月體形成的獨立危險因素,提示臨床醫(yī)師對有上述指標異常時應警惕新月體形成可能,并給予積極治療,以減輕腎臟損害。 第二部分MiR-29b在血管緊張素Ⅱ誘導的系膜細胞活化中的作用 目的探討miR-29b在血管緊張素Ⅱ(AngiotensinⅡ,AngⅡ)介導的腎小球系膜細胞活化(增殖和炎癥介質(zhì)表達)中的作用及其機制。 方法體外培養(yǎng)人類腎小球系膜細胞(Human glomerular mesangial cells,HGMCs),,100nmol/L AngⅡ刺激HGMCs;或穩(wěn)定轉(zhuǎn)染LV-hsa-miR-29b-1載體過表達miR-29b;或穩(wěn)定轉(zhuǎn)染LV-hsa-miR-29b-3p-inhibition載體抑制miR-29b表達后再應用AngⅡ刺激。采用real-time PCR和western blot檢測miR-29b、細胞間粘附分子-1(intracellular adhesion molecule-1,ICAM-1)、白介素-1β(interleukin-1β,IL-1β)、IL-6、IL-8、腫瘤壞死因子α誘導蛋白質(zhì)3(tumor necrosisfactor, alpha-induced protein3,TNFAIP3)、NF-κB抑制因子(NF-κB repressionfactor,NKRF)、細胞周期蛋白(CyclinA2、CyclinD1)的表達;ELISA檢測IL-1β、IL-8的表達;流式細胞術(shù)檢測細胞周期;CCK8法檢測細胞增殖。 結(jié)果(1)AngⅡ刺激后miR-29b、炎癥介質(zhì)ICAM-1、IL-1β、IL-6、IL-8及細胞周期蛋白CyclinA2、CyclinD1的表達顯著增加,細胞周期中S+G2/M期的比例增加(P0.05)。(2)miR-29b過表達誘導炎癥介質(zhì)ICAM-1、IL-1β、IL-6、IL-8的釋放、細胞周期蛋白CyclinA2、CyclinD1的表達增加及細胞增殖(P0.05),并且抑制靶基因TNFAIP3、NKRF的表達。(3)miR-29b表達抑制阻斷AngⅡ誘導的炎癥介質(zhì)釋放、細胞周期蛋白表達增加及細胞增殖(P0.05),并且誘導靶基因TNFAIP3、NKRF的表達增加。 結(jié)論miR-29b通過抑制靶基因TNFAIP3和NKRF的表達而促進炎癥介質(zhì)的釋放,通過調(diào)節(jié)細胞周期蛋白而促進HGMCs增殖。
[Abstract]:Expression of MiR - 29b in renal tissue of Henoch - Schonlein purpura nephritis
Objective To investigate the expression of miR - 29b in renal tissue of Henoch Schnlein purpura nephritis ( HSPN ) and to explore its relationship with HSPN pathological injury .
Methods The expression level of miR - 29b in renal biopsy was measured by real - time PCR , and the difference between the expression level of miR - 29b and the laboratory indexes was analyzed by means of real - time PCR . The relationship between the expression level of miR - 29b and the laboratory indexes was analyzed by SPSS 17.0 software , and the relationship between the expression level of miR - 29b and the new moon body of HSPN was further analyzed .
Results ( 1 ) The expression level of miR - 29b was increased ( P0.05 ) , and the expression level of miR - 29b in other pathological changes was not changed . ( 2 ) There was statistical difference between the two groups ( P0.05 ) .
Conclusion The expression level of miR - 29b in HSPN renal tissue with new moon body is increased . miR - 29b , 24hUPro / kg and URBC are independent risk factors formed in the crescent body of Henoch - Schonlein purpura nephritis .
Role of the second part MiR - 29b in the activation of angiotensin II - induced mesangial cells
Objective To investigate the role and mechanism of miR - 29b in the activation of mesangial cells ( proliferation and inflammatory mediators ) mediated by angiotensin II ( Ang 鈪
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