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微小RNA-128介导大麻素受体1(CB1)的表达,并调节核因子κB/磷酸化应激活化蛋白激酶(NF-κB/p-JNK)信号轴,以影响糖尿病膀胱病的发生。

microRNA-128 mediates CB1 expression and regulates NF-KB/p-JNK axis to influence the occurrence of diabetic bladder disease.

作者信息

Gou Xin, Wu Jing, Huang Mingqing, Weng Yuting, Yang Tongxin, Chen Tao, Li Guiqing, Fang Kewei

机构信息

Department of Urology, The Second Affiliated Hospital of Kunming Medical University, No. 374, Dianmian Dadao, Kunming, Yunnan, 650101, People's Republic of China.

Department of Biochemistry and Molecular Biology, The Primary Medicine School of Kunming Medical University, Kunming, 650101, People's Republic of China.

出版信息

J Transl Med. 2020 Jul 16;18(1):284. doi: 10.1186/s12967-020-02406-9.

Abstract

BACKGROUND

Diabetic bladder disease is common complications of diabetes, its symptoms are diverse, can be due to different stages. In this study we investigate the mechanism of miR-128 targeting CB1 expression to mediate the occurrence of diabetic bladder disease.

METHODS

Bioinformatics analysis predicts related regulatory factors of miR-128 in diabetic bladder disease. Models of diabetic bladder lesions were constructed in male SD rats by intraperitoneal injection of streptozotocin at 65 mg/kg body weight. The expression of miR-128 and CB1 mRNA in bladder tissues of each group was detected by RT-qPCR, and CB1, NF-KB, p-JNK and Bcl2 protein expression was detected by Western Blotting. We tested the function of the bladder by urodynamics, detected the pathological characteristics of the bladder tissue by HE staining, and verified the targeting relationship between miR-128 and CB1 through the prediction of the biological website, dual luciferase reporter gene assay and RIP.

RESULTS

miR-128 was highly expressed in the bladder tissue of diabetic rats. Inhibition of miR-128 could improve the occurrence of diabetic bladder lesions in rats. miR-128 could target the inhibition of CB1 expression, and high expression of CB1 could antagonize miR-128 against diabetic bladder. In the diabetic bladder, miR-128 can regulate the expression of NF-KB and p-JNK through CB1 and affect the level of apoptosis. miR-128 regulates NF-KB/p-JNK through CB1, thus affecting the occurrence of diabetic bladder disease.

CONCLUSION

The high expression of miR-128 can down-regulate the expression of CB1, promote the activation of NF-KB and p-JNK, increase the level of apoptosis and promote the occurrence of diabetic bladder disease.

摘要

背景

糖尿病膀胱病是糖尿病常见并发症,其症状多样,可因不同阶段而异。本研究旨在探讨miR-128靶向CB1表达介导糖尿病膀胱病发生的机制。

方法

生物信息学分析预测miR-128在糖尿病膀胱病中的相关调控因子。通过腹腔注射65mg/kg体重的链脲佐菌素在雄性SD大鼠中构建糖尿病膀胱病变模型。采用RT-qPCR检测各组膀胱组织中miR-128和CB1 mRNA的表达,采用蛋白质免疫印迹法检测CB1、NF-κB、p-JNK和Bcl2蛋白表达。通过尿动力学检测膀胱功能,通过HE染色检测膀胱组织病理特征,并通过生物网站预测、双荧光素酶报告基因检测和RNA免疫沉淀验证miR-128与CB1的靶向关系。

结果

miR-128在糖尿病大鼠膀胱组织中高表达。抑制miR-128可改善大鼠糖尿病膀胱病变的发生。miR-128可靶向抑制CB1表达,CB1高表达可拮抗miR-128对糖尿病膀胱的作用。在糖尿病膀胱中,miR-128可通过CB1调节NF-κB和p-JNK的表达并影响细胞凋亡水平。miR-128通过CB1调节NF-κB/p-JNK,从而影响糖尿病膀胱病的发生。

结论

miR-128高表达可下调CB1表达,促进NF-κB和p-JNK激活,增加细胞凋亡水平,促进糖尿病膀胱病的发生。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/631b/7367232/e024335ff9ed/12967_2020_2406_Fig1_HTML.jpg

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