Zhang Weichang, Yan Cheng, Xiao Yangyan, Sun Yuxin, Lin Youjun, Li Qinglong, Cai Wenwu
Department of Vascular Surgery, Second Xiangya Hospital, Central South University, Changsha, China.
Department of General Surgery, Second Xiangya Hospital, Central South University, Changsha, China.
Front Bioeng Biotechnol. 2023 May 23;11:1199785. doi: 10.3389/fbioe.2023.1199785. eCollection 2023.
Neointimal hyperplasia (NH) is a crucial pathophysiological feature in vascular transplant and in-stent restenosis. Excessive proliferation and migration of vascular smooth muscle cells (VSMCs) play important roles in neointimal hyperplasia. This study aims to explore the potentialities and mechanism of sulfasalazine (SSZ) in the prevention of restenosis. Sulfasalazine was encapsulated in nanoparticles made of poly (lactic-co-glycolic acid) (PLGA). , carotid ligation injury was induced in mice to induce Neointimal hyperplasia, with or without sulfasalazine containing nanoparticles (NP-SSZ) treatment. After 4 weeks, the arteries were collected for histology, immunofluorescence, Western blotting (WB) and qRT-PCR. , vascular smooth muscle cells were treated with TNF-α to induce cell proliferation and migration, followed by SSZ or vehicle treatment. WB was performed to further explore its mechanism. The ratio of intima to media thickness (I/M) was increased after ligation injury on day 28, while the ratio was significantly reduced in the NP-SSZ treatment group. The dual positive nuclei of Ki-67 and α-SMA were 47.83% ± 9.15%, whereas only 29.83% ± 5.98% in the NP-SSZ-treated group ( < 0.05). Both MMP-2 and MMP-9 were decreased in the NP-SSZ treatment group ( < 0.05, < 0.05, respectively) compared to the control group. The levels of the targeted inflammatory genes (TNF-α, VCAM-1, ICAM-1, MCP-1) were lower in the NP-SSZ treatment group compared with the control group. , the proliferating cell nuclear antigen (PCNA) expression was significantly decreased in the SSZ treatment group. The cell viability of VSMCs was markedly increased in the TNF-α treatment group, whereas sulfasalazine treatment inhibited this effect. LC3 II and P62 protein expression were higher in the SSZ group than in the vehicle group both and . The phosphorylation of NF-kB (p-NF-kB) and the phosphorylation of mTOR (p-mTOR) were decreased in the TNF-α+ SSZ group, whereas the P62 and LC3 II expression levels were increased. However, the expression level of p-mTOR, P62, and LC3 II was reversed after co-treatment with the agonist of mTOR MHY1485, whereas the p-NF-kB expression level was unchanged. sulfasalazine inhibited vascular smooth muscle cells proliferation and migration and Neointimal hyperplasia through NF-kB/mTOR-mediated autophagy.
新生内膜增生（NH）是血管移植和支架内再狭窄的关键病理生理特征。血管平滑肌细胞（VSMC）的过度增殖和迁移在新生内膜增生中起重要作用。本研究旨在探讨柳氮磺胺吡啶（SSZ）预防再狭窄的潜力和机制。柳氮磺胺吡啶被包裹在聚（乳酸-乙醇酸）（PLGA）制成的纳米颗粒中。通过对小鼠进行颈动脉结扎损伤以诱导新生内膜增生，分别给予或不给予含柳氮磺胺吡啶的纳米颗粒（NP-SSZ）治疗。4周后，收集动脉进行组织学、免疫荧光、蛋白质印迹法（WB）和定量逆转录聚合酶链反应（qRT-PCR）检测。此外，用肿瘤坏死因子-α（TNF-α）处理血管平滑肌细胞以诱导细胞增殖和迁移，随后给予SSZ或溶剂处理。通过WB进一步探讨其机制。结扎损伤后第28天，内膜与中膜厚度比（I/M）增加，而NP-SSZ治疗组该比值显著降低。增殖细胞核抗原（PCNA）和α-平滑肌肌动蛋白（α-SMA）双阳性核在对照组中为47.83%±9.15%，而在NP-SSZ治疗组中仅为29.83%±5.98%（P<0.05）。与对照组相比，NP-SSZ治疗组基质金属蛋白酶-2（MMP-2）和基质金属蛋白酶-9（MMP-9）均降低（分别为P<0.05，P<0.05）。NP-SSZ治疗组中靶向炎症基因（TNF-α、血管细胞黏附分子-1、细胞间黏附分子-1、单核细胞趋化蛋白-1）水平低于对照组。此外，SSZ治疗组中PCNA表达显著降低。TNF-α处理组血管平滑肌细胞活力显著增加，而柳氮磺胺吡啶处理可抑制此效应。在处理前和处理后，SSZ组中微管相关蛋白1轻链3Ⅱ（LC3Ⅱ）和P62蛋白表达均高于溶剂组。TNF-α+SSZ组中核因子-κB（NF-κB）磷酸化（p-NF-κB）和哺乳动物雷帕霉素靶蛋白（mTOR）磷酸化（p-mTOR）降低，而P62和LC3Ⅱ表达水平升高。然而，与mTOR激动剂MHY1485共同处理后，p-mTOR、P62和LC3Ⅱ表达水平逆转，而p-NF-κB表达水平未改变。柳氮磺胺吡啶通过NF-κB/mTOR介导的自噬抑制血管平滑肌细胞增殖、迁移及新生内膜增生。
