Down-regulation of miR-34a alleviates mesangial proliferation in vitro and glomerular hypertrophy in early diabetic nephropathy mice by targeting GAS1.
Down-regulation of miR-34a alleviates mesangial proliferation in vitro and glomerular hypertrophy in early diabetic nephropathy mice by targeting GAS1.
复制标题
DOI：
10.1016/j.jdiacomp.2014.01.002
复制
发表时间：
2014-05
影响因子：
3
通讯作者：
Zhang K
中科院分区：
文献类型：
--
作者：
Zhang L;He S;Guo S;Xie W;Xin R;Yu H;Yang F;Qiu J;Zhang D;Zhou S;Zhang K
糖尿病肾病是以肾小球系膜增生和肾小球肥大为主要特征的糖尿病并发症。MicroRNAs可能在这些病理过程中发挥重要作用。本研究的目的是研究miR-34a作为microRNAs之一与糖尿病肾病的可能联系及其在体内外的潜在机制。根据已有的比较糖尿病小鼠和正常对照小鼠不同microRNAs的芯片结果，选择miR-34a并用qRT-PCR检测其表达。用细胞计数试剂盒(CCK8)和5-乙炔基-20-脱氧尿苷(EDU)掺入法检测细胞存活率。Db/db小鼠注射安泰可下调miR-34a的表达。肾组织行高碘酸席夫(PAS)染色，计算肾小球平均直径。用荧光素酶基因报告法鉴定miR-34a的目的基因。免疫印迹和免疫组织化学分析验证生长停滞特异性1(GAS1)的表达水平。体外和体内高糖条件下，miR-34a的表达水平均升高。下调miR-34a在体外可抑制小鼠肾小球系膜细胞(MMCs)的增殖，在体内可减轻肾小球肥大。荧光素酶报告证实Gas1是miR-34a的靶标。此外，在高糖处理的MMCs和db/db小鼠中，分别观察到存在miR-34a反义核酸的MMCs和db/db小鼠的GAS1表达上调。在早期糖尿病肾病中，MIR-34a通过直接抑制GAS1而调节系膜细胞增殖和肾小球肥大。
Diabetic nephropathy (DN) is a major diabetic complication characterized by mesangial proliferation and glomerular hypertrophy. MicroRNAs might play an important role in these pathological processes. The aim of this study is to examine the possible association of miR-34a as one of the microRNAs with DN and underlying mechanisms in vitro and in vivo. According to previous results of microarray which compared the different microRNAs between diabetic and normal control mice, miR-34a was chosen and its expression was detected by qRT-PCR. Cell viability was then assessed using Cell Counting Kit-8 (CCK8) and 5-ethynyl-20-deoxyuridine (EDU) incorporation. Antagomir was injected in db/db mice to down regulate miR-34a. Average diameter of glomeruli was analyzed by periodic acid-Schiff (PAS) stain of kidney. Luciferase gene report assay was then performed to identify the target gene of miR-34a. Additional immunoblotting and immunohistochemical analyses were implemented to verify the expression level of growth arrest-specific 1 (GAS1). MiR-34a expression level was increased under high glucose condition in vitro and in vivo. Down-regulation of miR-34a inhibits mice mesangial cells (MMCs) proliferation in vitro and alleviates glomerular hypertrophy in vivo. GAS1 was proved to be the target of miR-34a through luciferase report. Moreover, up-regulation of GAS1 expression was observed in the presence of miR-34a antagomir as compared withmiR-34a antagomir-NC in high-glucose-treated MMCs and db/db mice, respectively. MiR-34a regulated mesangial proliferation and glomerular hypertrophy by directly inhibiting GAS1 in early DN.
登录
查看更多内容
DOI：
10.1073/pnas.1206432109
发表时间：
2012-10-23
影响因子：
11.1
作者：
Bernardo, Bianca C.;Gao, Xiao-Ming;McMullen, Julie R.
通讯作者：
McMullen, Julie R.
DOI：
10.1016/j.bbrc.2004.02.175
发表时间：
2004-04-23
影响因子：
3.1
作者：
Onozaki, A;Midorikawa, S;Watanabe, T
通讯作者：
Watanabe, T
DOI：
10.1016/j.biocel.2012.01.001
发表时间：
2012-04-01
影响因子：
4
作者：
Zhang, Lanyu;Pang, Shanshan;Le, Yingying
通讯作者：
Le, Yingying
影响因子：
3.5
作者：
Stebel, M;Vatta, P;Schneider, C
通讯作者：
Schneider, C
影响因子：
3.7
作者：
Chen H;Sun Y;Dong R;Yang S;Pan C;Xiang D;Miao M;Jiao B
通讯作者：
Jiao B