Homocysteine induces human mesangial cell apoptosis via the involvement of autophagy and endoplasmic reticulum stress.
Homocysteine induces human mesangial cell apoptosis via the involvement of autophagy and endoplasmic reticulum stress.
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同型半胱氨酸通过自噬和内质网应激诱导人系膜细胞凋亡
作者：
同型半胱氨酸（Hcy）水平在慢性肾脏病（CKD）中呈进行性升高的特征。事实上，Hcy积累被认为是CKD进展的关键生化罪魁祸首，但其潜在机制仍知之甚少。本研究探讨了同型半胱氨酸在肾小球系膜细胞（MC）凋亡中的作用以及自噬和内质网（ER）应激在这一过程中的潜在参与，揭示了同型半胱氨酸在肾脏疾病中的毒性作用。将人肾小球系膜细胞（HMCs）与不同浓度的同型半胱氨酸（Hcy）孵育不同时间。流式细胞术用于确定凋亡细胞的比例，蛋白质印迹法用于分析同型半胱氨酸，内质网抑制剂4-苯基丁酸（4-PBA）和Atg5 siRNA给药后的蛋白质水平。结果表明，随着Hcy浓度的增加和孵育时间的延长，HMCs的存活率逐渐降低，凋亡细胞比例逐渐增加。与此同时，凋亡相关蛋白Bax和切割的caspase-3的水平显著增加，而ER应激相关蛋白如ATF4、CHOP、GRP78和磷酸化eIF2 α的水平显著增加。裂解的LC 3、beclin 1和Atg 5蛋白水平也增加，伴随着p62降解，表明自噬激活。4-PBA能有效抑制内质网应激，逆转Hcy诱导的细胞凋亡和自噬。Atg5 siRNA可减轻Hcy诱导的细胞凋亡。综上所述，这些结果表明，同型半胱氨酸诱导HMC凋亡的剂量和时间依赖性的方式通过激活Atg5依赖性自噬触发ER应激。这项研究提出了一种新的策略，对同型半胱氨酸毒性肾损伤，并应有助于澄清CKD的发病机制。
Homocysteine (Hcy) level characterizes a progressive increase in chronic kidney disease (CKD). In fact, Hcy accumulation is considered to be a crucial biochemical culprit in CKD progression, but the mechanism underlying this remains poorly understood. This study investigated the role of Hcy in glomerular mesangial cell (MC) apoptosis and the potential involvement of autophagy and endoplasmic reticulum (ER) stress in this process, shedding light on Hcy toxicity in kidney disease. Human mesangial cells (HMCs) were incubated with different concentrations of Hcy for different times. Flow cytometry was used to determine the proportion of apoptotic cells and western blotting was used to analyze protein levels after the administration of Hcy, endoplasmic reticulum inhibitor 4-phenylbutyric acid (4-PBA), and Atg5 siRNA. The results demonstrated that the cell viability gradually decreased and the proportion of HMCs undergoing apoptosis increased with increasing Hcy concentration and prolonged incubation time. Meanwhile, levels of the apoptosis-related proteins Bax and cleaved caspase-3 were significantly increased, while ER stress-related proteins such as ATF4, CHOP, GRP78, and phospho-eIF2α significantly increased. Levels of cleaved LC3, and beclin1 and Atg5 proteins also increased, accompanied by p62 degradation, indicating autophagy activation. 4-PBA effectively inhibited ER stress and reversed Hcy-induced apoptosis and autophagy. Moreover, Atg5 siRNA alleviated Hcy-induced apoptosis. Taken together, these results suggest that Hcy induces HMC apoptosis in a dose- and time-dependent manner via the activation of Atg5-dependent autophagy triggered by ER stress. This study suggests a novel strategy against Hcy toxicity in kidney injury and should help in clarifying the pathogenesis of CKD.
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9
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