摘要
探究山药总蛋白对高浓度D-葡萄糖(High concentrations of D-glucose,HG)诱导的人脐静脉内皮细胞(Human umbilical vein endothelial cells,HUVECs)氧化应激损伤的保护作用及其机制.利用50 mmol/L HG建立HUVECs损伤模型.将HUVECs随机分为对照组、模型组、山药总蛋白低剂量组(0.5mg/mL)以及山药总蛋白高剂量组(1 mg/mL),评估山药总蛋白对HG处理的HUVECs的细胞活力、细胞形态及血管生成能力的影响;检测细胞中活性氧(Reactive oxygen species,ROS)的水平和超氧化物歧化酶(Superoxide dismutase,SOD)、过氧化氢酶(Catalase,CAT)的活力;以及培养基上清液中一氧化氮(Nitric oxide,NO)、8-羟基脱氧鸟苷(8-Hydroxy deoxyguanosine,8-OHdG)、丙二醛(Malondialdehyde,MDA)、白介素-1β(Interleukin-1β,IL-1β)、白介素-6(Interleukin-6,IL-6)的水平;测定B细胞淋巴瘤2(Bcl-2)、Bcl-2相关X蛋白(Bax)、硫氧还蛋白相互作用蛋白(Txnip)、NOD样受体蛋白3(NLRP3)、半胱氨酸天冬氨酸蛋白水解酶剪切体-1(Cleaved caspase-1)和IL-1β蛋白的表达水平.结果显示,山药总蛋白能显著提高HG处理后的HUVECs的细胞活力(P＜0.01),改善细胞凋亡形态,并显著升高SOD和CAT的活力(P＜0.05或P＜0.01)以及凋亡相关蛋白(Bcl-2/Bax)的比值、血管形成能力、NO分泌量.此外,山药总蛋白还显著降低了 HUVECs的ROS、MDA、8-OHdG、IL-1β 和 IL-6 的水平(P＜0.05 或P＜0.01)及炎症相关蛋白(Txnip、NLRP3、cleaved Caspase-1、IL-1β)的表达水平(P＜0.05或P＜0.01),并且上述指标的变化均呈现明显的浓度依赖性.本研究结果表明,山药总蛋白可能通过抑制Txnip/NLRP3信号通路从而保护HUVECs拮抗HG诱导的氧化应激,恢复HUVECs内的氧化平衡,进一步减轻细胞的炎症反应.
Abstract
To explored the protective effect and mechanism of Chinese yam total protein in human umbilical vein endothelial cells(HUVECs)with high-concentration D-glucose(HG)-induced oxidative stress injury.The HUVECs injury model was established by 50 mmol/L HG.HUVECs were randomly divided into control,model,low-dose Chinese yam total protein(0.5 mg/mL),and high-dose Chinese yam total protein(1 mg/mL)groups.The effects of Chinese yam total protein on the viability,morphology,and angiogenetic ability of HUVECs treated with HG were evaluated.The reactive oxygen species(ROS)level,superoxide dismutase(SOD)and catalase(CAT)activities in cells were determined.The nitric oxide(NO),8-hydroxy deoxyguanosine(8-OHdG),malondialdehyde(MDA),interleukin-1β(IL-1β),and interleukin-6(IL-6)levels in culture medium supernatant were measured.Protein expression levels of B-cell lymphoma 2(Bcl-2),Bcl-2-associated X(Bax),thioredoxin-interacting protein(Txnip),NOD-like receptor protein 3(NLRP3),cleaved Caspase-1,and IL-1β were determined.The results showed that Chinese yam total protein could significantly increased the cell viability of HUVECs treated with HG(P＜0.01),improved cell apoptotic morphology,and significantly increased the activities of SOD and CAT(P＜0.05,P＜0.01)and ratio of apoptosis-related proteins(Bcl-2/Bax),angiogenetic ability and nitric oxide(NO)secretion.Moreover,Chinese yam total protein significantly decreased the levels of ROS,MDA,8-OHdG,IL-1β,and IL-6(P＜0.05,P＜0.0 1)and the expression levels of inflammation-related proteins(Txnip,NLRP3,cleaved Caspase-1,and IL-1β)(P＜0.05,P＜0.01).Changes in these indicators showed a significant concentration-dependent manner.The results indicate that Chinese yam total protein may protect against HG-induced oxidative stress by inhibiting the Txnip/NLRP3 signaling pathway,restoring the oxidative balance,and reducing the inflammatory response of HUVECs.
基金项目
吉林省科技发展计划(YDZJ202301ZYTS445)
万方数据