Department of Physiology, Institute of Basic Medicine, Hebei Medical University, 050017, Hebei, China; Department of Physiology, Institute of Basic Medicine, Xingtai Medical College, 054000, Hebei, China.
Department of Physiology, Institute of Basic Medicine, Hebei Medical University, 050017, Hebei, China.
Nitric Oxide. 2024 Dec 1;153:72-81. doi: 10.1016/j.niox.2024.10.008. Epub 2024 Oct 11.
Previous studies showed that hyperhomocysteinemia (HHcy) induced endothelial dysfunction by endoplasmic reticulum (ER) stress induction and autophagy stimulation. This study aimed to determine the effect of hydrogen sulfide (HS) in homocysteine (Hcy)-induced endothelial dysfunction and observe the possible mechanism involved. Male Wistar rats (160-180g) were used and randomly divided into four groups: Control group, HHcy group, HHcy+Sodium hydrosulfide (NaHS) group and NaHS group. Rats were fed with 2% high methionine diet for 8 weeks to set up HHcy model. Plasma concentration of Hcy was measured by ELISA. Endothelium-dependent and non-endothelium-dependent vasodilation of rat renal arteries were determined by myograph. The protein expression of cystathionine-γ-lyase (CSE), ER stress- and autophagy-related proteins in renal arteries or human umbilical vein endothelial cells (HUVECs) were analyzed by western blotting. The endothelial function was impaired in HHcy rats and HUVECs. NaHS supplementation could improve the ACh-induced vasodilation, however it was eliminated by ER stress inducer Tunicamycin (TM) or autophagy inducer Rapamycin. Western blotting in renal arteries showed that Glucose-regulated protein 78 (GRP78) and three branches of ER stress (p-IRE1α, p-PERK, ATF6) , p-JNK1+p-JNK2 were downregulated, simultaneously the autophagy marker Beclin1, LC3BII/LC3BI ratio were decreased and p62 was increased with NaHS treatment in HHcy rats. In HUVECs, IRE1α-JNK induced autophagy was involved in HHcy-induced endothelial dysfunction, while NaHS stimulation reversed the protein expression in IRE1α/JNK-autophagy pathway with Hcy incubation. This study might suggest that endothelial dysfunction induced by HHcy might be correlated with IRE1α-JNK-autophagy axis pathway, which was suppressed by exogenous supplementation of HS donor, NaHS.
先前的研究表明，高同型半胱氨酸血症（HHcy）通过内质网（ER）应激诱导和自噬刺激引起内皮功能障碍。本研究旨在确定硫化氢（HS）对同型半胱氨酸（Hcy）诱导的内皮功能障碍的影响，并观察可能涉及的机制。雄性 Wistar 大鼠（160-180g）被用于并随机分为四组：对照组、HHcy 组、HHcy+硫氢化钠（NaHS）组和 NaHS 组。大鼠用 2%高蛋氨酸饮食喂养 8 周建立 HHcy 模型。用 ELISA 法测定血浆 Hcy 浓度。用肌动描记法测定大鼠肾动脉的内皮依赖性和非内皮依赖性血管舒张。用 Western blot 分析肾动脉或人脐静脉内皮细胞（HUVEC）中胱硫醚-γ-裂解酶（CSE）、ER 应激和自噬相关蛋白的蛋白表达。HHcy 大鼠和 HUVEC 内皮功能受损。NaHS 补充可改善 ACh 诱导的血管舒张，但被 ER 应激诱导剂衣霉素（TM）或自噬诱导剂雷帕霉素消除。肾动脉的 Western blot 显示葡萄糖调节蛋白 78（GRP78）和 ER 应激的三个分支（p-IRE1α、p-PERK、ATF6）、p-JNK1+p-JNK2 下调，同时自噬标志物 Beclin1、LC3BII/LC3BI 比值降低，p62 升高与 HHcy 大鼠的 NaHS 治疗相关。在 HUVEC 中，IRE1α-JNK 诱导的自噬参与 HHcy 诱导的内皮功能障碍，而 Hcy 孵育时 NaHS 刺激逆转了 IRE1α/JNK-自噬通路的蛋白表达。本研究表明，HHcy 诱导的内皮功能障碍可能与 IRE1α-JNK-自噬轴途径相关，该途径受外源性 HS 供体 NaHS 的抑制。
