中华生物医学工程杂志
中華生物醫學工程雜誌
중화생물의학공정잡지
CHINESE JOURNAL OF BIOMEDICAL ENGINEERING
2009年
4期
239-243
,共5页
蒋小云%吴伟%张巧玲%莫樱%陈丽植%陆慧瑜%陈述枚
蔣小雲%吳偉%張巧玲%莫櫻%陳麗植%陸慧瑜%陳述枚
장소운%오위%장교령%막앵%진려식%륙혜유%진술매
肾小球肾炎%IgA%肾小球系膜细胞%转化生长因子β1%磷酸化Smad3%纤连蛋白
腎小毬腎炎%IgA%腎小毬繫膜細胞%轉化生長因子β1%燐痠化Smad3%纖連蛋白
신소구신염%IgA%신소구계막세포%전화생장인자β1%린산화Smad3%섬련단백
Glomerulonephritis%IgA%Mesangial cells%Transforming growth factor beta1%Phosphorylated Smad3%Fibronectins
目的 研究IgA肾病(IgAN)患者血清IgA1对人肾小球系膜细胞(HMC)转化生长因子β1(TGF-β1)、磷酸化Smad3(p-Smad3)和纤连蛋白(FN)的刺激作用,探讨IgA1对TGF-β1/Smads信号通路的影响.方法 体外培养HMC,分5组.即健康对照组、健康人IgA1(NIgA1)刺激组、健康人聚合IgA1(aNIgA1)刺激组、IgAN患者IgA1(PIgA1)刺激组和IgAN患者聚合IgA1(aPIgA1)刺激组.RT-PCR检测各组细胞TGF-β1、p-Smad3和FNmRNA的表达.Western免疫印迹检测HMC p-Smad3蛋白水平.ELISA检测HMC培养上清液中TGF-β1、FN蛋白水平.结果 PIgA1和aPIgA1能显著上调TGF-β1,P-Smad3、FN mRNA和蛋白的表达(P<0.05).aPIgA1上调三者mRNA表达的能力分别为PIgA1的1.5倍、1.3倍和1.5倍(均P<0.05).aPIgA1上调TGF-β1和p-Smad3蛋白表达的能力分别为PIgA1的2.1倍和1.6倍(均P<0.01).在aPIgA1刺激不同时间,TGF-β1和p-Smad3 mRNA表达于12 h达高峰(0.866±0.055和0.891±0.251),于24h回到基线水平.FNmRNA表达于24 h达高峰(0.968±0.031).TGF-β1和p-Smad3蛋白表达逐渐升高,TGF-β1于12 h达高峰[(246.960±1.270)ng/L],p-Smad3于6 h达高峰(0.490±0.046)后开始下降.FN蛋白表达逐渐升高,24 h达高峰[(1.804±0.038)mg/L](均P<0.01).结论 PIgA1和aPIgA1能显著上调HMC TGF-β1、p-smad3、和FNmRNA表达和蛋白水平.且aPIgA1的作用强于PIgA1,提示IgAN患者血清的IgA1,主要是aIgA1可通过TGF-β1/Smads信号通路在IgAN进展中起作用.
目的 研究IgA腎病(IgAN)患者血清IgA1對人腎小毬繫膜細胞(HMC)轉化生長因子β1(TGF-β1)、燐痠化Smad3(p-Smad3)和纖連蛋白(FN)的刺激作用,探討IgA1對TGF-β1/Smads信號通路的影響.方法 體外培養HMC,分5組.即健康對照組、健康人IgA1(NIgA1)刺激組、健康人聚閤IgA1(aNIgA1)刺激組、IgAN患者IgA1(PIgA1)刺激組和IgAN患者聚閤IgA1(aPIgA1)刺激組.RT-PCR檢測各組細胞TGF-β1、p-Smad3和FNmRNA的錶達.Western免疫印跡檢測HMC p-Smad3蛋白水平.ELISA檢測HMC培養上清液中TGF-β1、FN蛋白水平.結果 PIgA1和aPIgA1能顯著上調TGF-β1,P-Smad3、FN mRNA和蛋白的錶達(P<0.05).aPIgA1上調三者mRNA錶達的能力分彆為PIgA1的1.5倍、1.3倍和1.5倍(均P<0.05).aPIgA1上調TGF-β1和p-Smad3蛋白錶達的能力分彆為PIgA1的2.1倍和1.6倍(均P<0.01).在aPIgA1刺激不同時間,TGF-β1和p-Smad3 mRNA錶達于12 h達高峰(0.866±0.055和0.891±0.251),于24h迴到基線水平.FNmRNA錶達于24 h達高峰(0.968±0.031).TGF-β1和p-Smad3蛋白錶達逐漸升高,TGF-β1于12 h達高峰[(246.960±1.270)ng/L],p-Smad3于6 h達高峰(0.490±0.046)後開始下降.FN蛋白錶達逐漸升高,24 h達高峰[(1.804±0.038)mg/L](均P<0.01).結論 PIgA1和aPIgA1能顯著上調HMC TGF-β1、p-smad3、和FNmRNA錶達和蛋白水平.且aPIgA1的作用彊于PIgA1,提示IgAN患者血清的IgA1,主要是aIgA1可通過TGF-β1/Smads信號通路在IgAN進展中起作用.
목적 연구IgA신병(IgAN)환자혈청IgA1대인신소구계막세포(HMC)전화생장인자β1(TGF-β1)、린산화Smad3(p-Smad3)화섬련단백(FN)적자격작용,탐토IgA1대TGF-β1/Smads신호통로적영향.방법 체외배양HMC,분5조.즉건강대조조、건강인IgA1(NIgA1)자격조、건강인취합IgA1(aNIgA1)자격조、IgAN환자IgA1(PIgA1)자격조화IgAN환자취합IgA1(aPIgA1)자격조.RT-PCR검측각조세포TGF-β1、p-Smad3화FNmRNA적표체.Western면역인적검측HMC p-Smad3단백수평.ELISA검측HMC배양상청액중TGF-β1、FN단백수평.결과 PIgA1화aPIgA1능현저상조TGF-β1,P-Smad3、FN mRNA화단백적표체(P<0.05).aPIgA1상조삼자mRNA표체적능력분별위PIgA1적1.5배、1.3배화1.5배(균P<0.05).aPIgA1상조TGF-β1화p-Smad3단백표체적능력분별위PIgA1적2.1배화1.6배(균P<0.01).재aPIgA1자격불동시간,TGF-β1화p-Smad3 mRNA표체우12 h체고봉(0.866±0.055화0.891±0.251),우24h회도기선수평.FNmRNA표체우24 h체고봉(0.968±0.031).TGF-β1화p-Smad3단백표체축점승고,TGF-β1우12 h체고봉[(246.960±1.270)ng/L],p-Smad3우6 h체고봉(0.490±0.046)후개시하강.FN단백표체축점승고,24 h체고봉[(1.804±0.038)mg/L](균P<0.01).결론 PIgA1화aPIgA1능현저상조HMC TGF-β1、p-smad3、화FNmRNA표체화단백수평.차aPIgA1적작용강우PIgA1,제시IgAN환자혈청적IgA1,주요시aIgA1가통과TGF-β1/Smads신호통로재IgAN진전중기작용.
Objective To explore the effect of serum IgA1 from patients with IgA nephropathy(IgAN) on transforming growth factor beta- 1 ( TGF-β 1 ), phosphorylated Smad3 (p-Smad3) and fibronectin(FN) in human glomerular mesangial cells (HMC). Methods HMC was cultured in vitro and divided into 5 groups: IgA1 free control (health control group), stimulated with serum IgA1 from healthy people (NIgA1group) , stimulated with serum aggregated IgA1 from healthy people (aNIgA1 group) , stimulated with serum IgA1 from patients with IgA nephropathy (IgAN) (PIgA1 group), and stimulated with serum aggregated IgA 1 from patients with IgAN (aPIgA1 group). The expression of TGF-β1, p-Smad3 and FN mRNA was assayed by RT-PCR. The expression of p-Smad3 protein was analyzed by Western blot. The expression of TGF-β1 and FN protein in the culture supematant of HMC was analyzed by ELISA. Results PIgA1 and aPIgA1 significantly up-regnlated TGF-β1, p-Smad3, FN mRNA and protein expression. The TGF-β1, p-Smad3 and FN mRNA were increased by 1.5-fold(P<0.01), 1.3-fold(P<0.05) and 1.5-fold(P< 0.01 ) in aPIgA1 group as compared with PIgA1 group. The TGF-β1 and p-Smad3 protein were increased by 2.1-fold and 1.6-fold (P<0.01) in aPIgA1 group as compared with PIgA1 group. In aPIgA1 group, the expression of TGF-β1 and p-Smad3 mRNA with aPIgA1 was increased gradually, peaked at 12 h (0.866± 0.055,0.891±0.251 ), and then decreased to the basic levels at 24 h, but the the expression of FN mRNA peaked at 24 h(0.968±0.031 ). The expression of TGF-β1 and p-Smad3 protein was increased , peaked at 12 h for TGF-β1 [(246.960±1.270) ng/L], at 6 h for p-Smad3(0.490±0.046), then decreased. The expression of FN protein was increased , peaked at 24 h [(1.804±0.038) mg/L] (P<0.01). Conclusions PIgA1 and aPIgA1 significantly up-regulate TGF-β, p-Smad3, FN mRNA and protein expression. The effect of aPIgA1 is stronger than that of PIgA1. It is suggested that serum IgA1, mainly aIgA1 from patients with IgAN may play an important role in the progression of IgAN through TGF-β1/Smads signaling pathway.
