Deng Jiuhong, Zheng Chao, Hua Zhou, Ci Haideng, Wang Guiying, Chen Lijing
Wenzhou Medical University, Chashan Higher Education Park, Wenzhou City, 325035, Zhejiang Province, China.
Department of Endocrinology, Second People's Hospital of Pingyang County, Wenzhou City, 325405, Zhejiang Province, China.
BMC Complement Med Ther. 2022 Apr 27;22(1):116. doi: 10.1186/s12906-022-03597-y.
Diosmin has been reported to treat diabetes, but its role in diabetic nephropathy (DN) remains unclear. This research investigated the mechanism by which diosmin alleviated high glucose (HG)-induced HK-2 cell injury.
First, we used CCK-8 to detect the effect of 0.1, 1, or 10 μg/mL diosmin on the viability of HK-2 cells treated with normal glucose or HG. Next, we used flow cytometry, automatic biochemical analyzer, ELISA, immunofluorescence, and colorimetric assay kit to examine the apoptosis, oxidative stress, inflammatory factors, and Caspase-3 expression in HK-2 cells. Thereafter, we used the western blot and qRT-PCR to examine the expression of the endoplasmic reticulum stress-, oxidative stress-, inflammation-, apoptosis-, and autophagy, and PI3K/AKT pathway-related factors.
Diosmin was non-cytotoxic to normal HK-2 cells and enhanced the HK-2 cell viability suppressed by HG. Meanwhile, diosmin restrained apoptosis, the contents of MDA, pro-inflammatory factors, and Caspase-3 but intensified the contents of SOD and CAT induced by HG. We further confirmed that diosmin blunted oxidative stress-, inflammation-, apoptosis-, and autophagy-related factors expression induced by HG via restraining the CHOP and GRP78 expressions. Further, we also discovered that PTEN level was restrained and the ratios of p-PI3K/PI3K and p-AKT/AKT were enhanced in HK-2 cells induced by HG, which was reversed by co-treatment of HG and diosmin.
Our study manifested that diosmin alleviated the HG-mediated endoplasmic reticulum stress injury in HK-2 cells via restraining the PI3K/AKT pathway.
据报道，地奥司明可治疗糖尿病，但其在糖尿病肾病（DN）中的作用仍不清楚。本研究探讨了地奥司明减轻高糖（HG）诱导的HK-2细胞损伤的机制。
首先，我们使用CCK-8检测0.1、1或10μg/mL地奥司明对正常葡萄糖或HG处理的HK-2细胞活力的影响。接下来，我们使用流式细胞术、自动生化分析仪、酶联免疫吸附测定、免疫荧光和比色测定试剂盒检测HK-2细胞中的凋亡、氧化应激、炎症因子和半胱天冬酶-3表达。此后，我们使用蛋白质免疫印迹法和定量逆转录聚合酶链反应检测内质网应激、氧化应激、炎症、凋亡和自噬以及PI3K/AKT通路相关因子的表达。
地奥司明对正常HK-2细胞无细胞毒性，并增强了被HG抑制的HK-2细胞活力。同时，地奥司明抑制凋亡、丙二醛含量、促炎因子和半胱天冬酶-3，但增强了HG诱导的超氧化物歧化酶和过氧化氢酶含量。我们进一步证实，地奥司明通过抑制CHOP和GRP78表达，减弱了HG诱导的氧化应激、炎症、凋亡和自噬相关因子表达。此外，我们还发现，HG诱导的HK-2细胞中PTEN水平受到抑制，p-PI3K/PI3K和p-AKT/AKT的比率增加，而HG与地奥司明共同处理可逆转这种情况。
我们的研究表明，地奥司明通过抑制PI3K/AKT通路减轻了HG介导的HK-2细胞内质网应激损伤。
