Alterations of urine microRNA-7977/G6PD level in patients with diabetic kidney disease and its association with dysfunction of albumin-induced autophagy in proximal epithelial tubular cells.

IF 4.2 2区 医学 Q1 ENDOCRINOLOGY & METABOLISM
Zhenzhen Shi, Xinran Li, Liyi Zhang, Jinlan Xie, Feifei Zhong, Zhenhong Guo, Zhongai Gao, Jingyu Wang, Roshan Kumar Mahto, Yuan Li, Shenglan Wang, Baocheng Chang, Robert C Stanton, Juhong Yang
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引用次数: 0

Abstract

Diabetic kidney disease (DKD) remains as one of the leading long-term complications of type 2 diabetic mellitus (T2DM). Studies have shown that decreased expression of glucose-6-phosphate dehydrogenase (G6PD) plays an important role in DKD. However, the upstream and downstream pathways of G6PD downregulation leading to DKD have not been elucidated. We conducted a series of studies including clinical study, animal studies, and in vitro studies to explore this. First, a total of 90 subjects were evaluated including 30 healthy subjects, 30 patients with T2DM, and 30 patients with DKD. The urinary G6PD activity and its association with the clinical markers were analyzed. Multivariate linear regression analysis was used to analyze the risk factors of urinary G6PD in these patients. Then, microRNAs that were differentially expressed in urine and could bind and degrade G6PD were screened and verified in patients with DKD. After that, high glucose (HG)-cultured human kidney cells (HK-2) and Zucker diabetic fatty (ZDF) rats were used to test the roles of miR-7977/G6PD/albumin-induced autophagy in DKD. Beclin and P62 were used as markers of kidney autophagy indicators. A dual-luciferase reporter assay system was used to test the binding of G6PD by mir-7977. The plasma and urinary G6PD activity were decreased significantly in patients with DKD, accompanied by increased urinary mir-7977 level. The fasting plasma glucose (FPG), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), and urinary albumin excretion were independent predictors of urinary G6PD activity, according to multiple linear regression analysis. The increased expression of miR-7977 and decreased expression of G6PD were also found in the kidney of ZDF rats with early renal tubular damage. The correlation analysis showed that beclin protein expression levels were positively correlated with kidney G6PD activity, whereas P62 protein expression was negatively correlated with kidney G6PD activity in rats. In HK-2 cells cultured with normal situation, a low level of albumin could induce autophagy along with the stimulation of G6PD, although this was impaired under high glucose. Overexpression of G6PD reversed albumin-induced autophagy in HK-2 cells under high glucose. Further study revealed that G6PD was a downstream target of miR-7977. Inhibition of miR-7977 expression led to significantly increased expression of G6PD and reversed the effects of high glucose on albumin-induced autophagy. In conclusion, our study supports a new mechanism of G6PD downregulation in DKD. Therapeutic measures targeting the miR-7977/G6PD/autophagy signaling pathway may help in the prevention and treatment of DKD.NEW & NOTEWORTHY This study provides new evidence that reduced glucose-6-phosphate dehydrogenase (G6PD) may damage the endocytosis of renal tubular epithelial cells by reducing albumin-induced autophagy. More importantly, for the first time, our study has provided evidence from humans that the decrease in urinary G6PD activity is positively associated with renal injury, and abnormal glucose and lipid metabolism may be important reasons for reduced G6PD levels. Increased miR-7977 may at least in part explain the downregulation of G6PD.

DKD患者尿液中的microRNA-7977/G6PD水平及其与近端上皮肾小管细胞白蛋白诱导的自噬功能障碍的关系。
研究表明,葡萄糖-6-磷酸脱氢酶(G6PD)表达下降在 DKD 中起着重要作用。我们对此进行了一系列研究,包括临床研究、动物实验和体外实验。首先,我们对 90 名受试者进行了评估。首先,我们对 90 名受试者进行了评估,分析了尿液中 G6PD 活性及其与临床指标的关系。然后,筛选并验证了 DKD 患者尿液中可结合和降解 G6PD 的不同 microRNA。随后,利用高糖(HG)培养的人肾细胞(HK-2)和扎克糖尿病脂肪大鼠(ZDF)检验了 miR-7977/G6PD/ 白蛋白诱导的自噬在 DKD 中的作用。DKD患者血浆和尿液中的G6PD活性明显降低,同时尿液中mir-7977水平升高。通过多元线性回归分析,空腹血浆葡萄糖(FPG)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)和尿白蛋白排泄量是尿液 G6PD 活性的独立预测因子。在正常情况下培养的 HK-2 细胞中,低水平的白蛋白可以诱导自噬,同时刺激 G6PD,但在高糖条件下这种作用会受到影响。抑制mir-7977的表达会导致G6PD的表达显著增加,并逆转高糖对白蛋白诱导的自噬的影响。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
CiteScore
9.80
自引率
0.00%
发文量
98
审稿时长
1 months
期刊介绍: The American Journal of Physiology-Endocrinology and Metabolism publishes original, mechanistic studies on the physiology of endocrine and metabolic systems. Physiological, cellular, and molecular studies in whole animals or humans will be considered. Specific themes include, but are not limited to, mechanisms of hormone and growth factor action; hormonal and nutritional regulation of metabolism, inflammation, microbiome and energy balance; integrative organ cross talk; paracrine and autocrine control of endocrine cells; function and activation of hormone receptors; endocrine or metabolic control of channels, transporters, and membrane function; temporal analysis of hormone secretion and metabolism; and mathematical/kinetic modeling of metabolism. Novel molecular, immunological, or biophysical studies of hormone action are also welcome.
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