摘要
目的 观察TRAM-34对TGF-β1诱导系膜细胞增生的影响,了解KCa 3.1在肾小球硬化中的作用.方法2ng/mL TGF-β1诱导系膜细胞增生后,采用流式细胞技术和MTT方法观察8、16、24nmol/L TRAM-34分别对细胞周期和细胞增殖的影响,并选用最佳抑制浓度TRAM-34干预15、30、60min后,用RT-PCR技术观察其对系膜细胞α-SMA和FSP-1转录水平的影响.结果 与空白对照组相比,2ng/mL TGF-β1诱导的系膜细胞阻滞于G0～G1期,表现为G0～G1期细胞百分比明显升高[（48.45±1.89）% vs.（70.29±1.84）%,P〈0.01],而S期细胞百分比相应下降[（38.54±1.13）% vs.（19.56±1.67）%;P〈0.01）;各浓度TRAM-34组均可以改善阻滞于G0～G1期的系膜细胞增生,使细胞周期中G0～G1期细胞百分比明显减少,S期细胞百分比明显增多,其中24nmol/L组降低最明显[TGF-β1组vs.TRAM-34组：G0～G1期为（70.29±1.84）% vs.（61.90±1.88）%;S期为（19.56±1.67）% vs.（25.52±1.62）%;P〈0.05）.MTT实验结果也显示各浓度TRAM-34对TGF-β1诱导系膜细胞增生具有抑制作用,其中24nmol/L组抑制作用最显著,与8nmol/L和16nmol/L组比较,差异有统计学意义（P〈0.01）.24nmol/L的TRAM-34可显著抑制系膜细胞α-SMA和FSP-1的表达.结论 TRAM-34能够改善TGF-β1诱导的系膜细胞阻滞,减少细胞的早衰、表型转化和纤维样增生.
Objective To investigate the effects of TRAM-34 on TGF-β1-induced mesangial cell hypertrophy to probe the role of Kca3.1 in glomerulosclerosis. Methods After 2 ng/mL TGF-β1 was used to induce mesangial cell hypertrophy/ proliferation, the effects of 8 nmol/L, 16 nmol/L and 24 nmol/L TRAM-34 upon cell cycle and cell proliferation were observed using flow cytometry and MTT. Then the most effective dosage of TRAM-a4 was identified and the mRNA of a-SMA and FSP-1 was tested by RT-PCR. Results The hypertrophy of mesangial cells induced by 2 ng/mL TGF-~I was blocked at G0--G1 phase control vs. TGF-β1. （48.45±1.89）% vs. （70.2±1.84）%; P〈0.01. At S phase, the percentage of cells declined correspondingly control vs. TGF-β1 ： （38.54± 1.13）% vs. （19.56±1.67）% ; P〈0.01. Different concentrations （8, 16 and 24 nmol/L） of TRAM-34 improved the mesangial cell arrest at G0--G1 phase. Of them, the effect was the most obvious in 24 nmol/L group TGF-β1 vs. TRAM-34; G0--G1 phase, （70.29±1.84）% vs. （61.90±1.88）%; Sphase, （19.56±1.67）% vs. （25.52±1.62） %（ P〈0.05. MTT assay showed the similar results that the inhibition in 24 nmol/L group was obviously higher than in 8 nmol/L and the 16 nmol/L groups （P〈0.01）. And the inhibitory effects of 24 nmol/L TRAM-34 on α-SMA and FSP-1 mRNA expressihons were also greater （P 〈 0.05）. Oorlcltlsiorl TRAM-34 may improve TGF-β1 -induced mesangial cell hypertrophy, cellular phenotype transformation and premature senescence.
出处
《西安交通大学学报（医学版）》
CAS
CSCD
北大核心
2014年第2期200-203,221,共5页
Journal of Xi’an Jiaotong University（Medical Sciences）
基金
国家自然科学基金资助项目(No.81100530
81070590
30801504
30901926)
陕西省科技攻关资助项目(No.2011K14-09-02
2012K16-08-04)~~
