CAJ | 학술논문

目的探讨内质网应激在高糖诱导人肾脏系膜细胞表型转化的作用.方法将体外培养的人肾脏系膜细胞分为对照组、高糖组(HG组)、高糖+4-苯墓丁酸(4-PBA)干预组(4-PBA组).MTF法测定细胞增殖率,流式细胞术观察细胞周期变化,免疫组织化学染色激光共聚焦显微镜观察细胞d平滑肌肌动蛋白(α-SMA)表达情况,实时荧光定量PCR和Westem印迹分别检测mRNA和蛋白表达.结果 (1)高糖组系膜细胞增殖率和进入S期比例显著高于对照组(P＜0.05),同时,高糖能刺激α-SMA表达增加(P＜0.05);4-PBA干预可明显抑制高糖刺激的系膜细胞增殖和进入S期及α-SMA表达.(2)相对于对照组,高糖能刺激系膜细胞葡萄糖调节蛋白78(Grp78)mRNA和蛋白表达增加(P＜0.05);4-PBA干预可显著下调这种效应(P＜0.05).(3)高糖可诱导系膜细胞转化生长因子(TGF-β1)、纤连蛋白(FN)mRNA和蛋白表达(P＜0.05);4-PBA干预则明显下调其表达(P＜0.05).结论内质网应激在高糖诱导人肾脏系膜细胞表型转化效应中发挥了重要作用.
목적 탐토내질망응격재고당유도인신장계막세포표형전화적작용.방법 장체외배양적인신장계막세포분위대조조、고당조(HG조)、고당+4-분묘정산(4-PBA)간예조(4-PBA조).MTF법측정세포증식솔,류식세포술관찰세포주기변화,면역조직화학염색격광공취초현미경관찰세포d평활기기동단백(α-SMA)표체정황,실시형광정량PCR화Westem인적분별검측mRNA화단백표체.결과 (1)고당조계막세포증식솔화진입S기비례현저고우대조조(P＜0.05),동시,고당능자격α-SMA표체증가(P＜0.05);4-PBA간예가명현억제고당자격적계막세포증식화진입S기급α-SMA표체.(2)상대우대조조,고당능자격계막세포포도당조절단백78(Grp78)mRNA화단백표체증가(P＜0.05);4-PBA간예가현저하조저충효응(P＜0.05).(3)고당가유도계막세포전화생장인자(TGF-β1)、섬련단백(FN)mRNA화단백표체(P＜0.05);4-PBA간예칙명현하조기표체(P＜0.05).결론 내질망응격재고당유도인신장계막세포표형전화효응중발휘료중요작용.
Objective To study the role of endoplasmic reticulum stress in phenotypic change of cultured human glomerular mesangial cells induced by high glucose.Methods Cultured human glomeruar mesangial cells were divided into three groups: control group,high glucose group and high glucose+ 4-phenylbutyric acid (4-PBA) group.Cell number of proliferation was assessed by MTT assay.Cell cycle was measured by flow cytometric analysis.Expression of α-SMA was assessed by immunohistochemistry and was observed by laser scanning confocal microscope.Involved mRNA and protein expression were measured by real-time PCR and Western blotting.Results (1)Cell number of proliferation and S transition proportion in high glucose group significantly increased than that in control group (P ＜ 0.05).High glucose could induce α-SMA expression significantly (P＜0.05).4-PBA could significantly inhibit human glomerular mesangial cells proliferation (P＜0.05),S transition arrest (P＜0.05) and expression of α-SMA (P＜0.05) induced by high glucose.(2) Compared with control group,high glucose could significantly increase the expression of glucose-regulated protein78(Grp78 ) mRNA and protein (P＜ 0.05),which could be inhibited by 4-PBA treatment (P＜0.05).(3)High glucose could induce the mRNA and protein expression of TGF-β1 and FN significantly,which could be inhibited by 4-PBA treatment (P＜0.05).Conclusion Endoplasmic reticulum stress plays an important role in phenotypic change of cultured human glomerular mesangial cells induced by high glucose.