Fu Dongdong, Wu Shuai, Jiang Xiangfu, You Tingyu, Li Yu, Xin Jiao, Feng Xiaowen, Wen Jiagen, Huang Yan, Hu Chengmu
Inflammation and Immune Mediated Diseases Laboratory of Anhui Province, Anhui Institute of Innovative Drugs, School of Pharmacy, Anhui Medical University, Hefei, 230032, China; Institute for Liver Diseases of Anhui Medical University, School of Pharmacy, Anhui Medical University, Hefei, 230032, China; Key Laboratory of Anti-inflammatory and Immune Medicine, Ministry of Education, Hefei, 230032, China.
Inflammation and Immune Mediated Diseases Laboratory of Anhui Province, Anhui Institute of Innovative Drugs, School of Pharmacy, Anhui Medical University, Hefei, 230032, China; Institute for Liver Diseases of Anhui Medical University, School of Pharmacy, Anhui Medical University, Hefei, 230032, China; Key Laboratory of Anti-inflammatory and Immune Medicine, Ministry of Education, Hefei, 230032, China.
Free Radic Biol Med. 2023 Feb 1;195:245-257. doi: 10.1016/j.freeradbiomed.2022.12.095. Epub 2022 Dec 31.
Acetaminophen (APAP) is one of the most widely used drugs in the world. The literature shows that excessive or long-term use of APAP can lead to increased cardiovascular dysfunction. An acute increase in angiotensin Ⅱ (Ang Ⅱ) caused by APAP use in fatty liver disease may increase the risk and severity of vascular injury. However, the underlying mechanism remains unclear. Caveolin-1 (CAV1) is a broad-spectrum kinase inhibitor that significantly determines endothelial function. This study aimed to observe the effects of APAP on the vasculature in non-alcoholic fatty liver disease (NAFLD) and to determine whether CAV1 could alleviate vascular oxidative stress and inflammation by targeting Ang Ⅱ or its downstream pathways. In this study, 7-week-old C57BL/6 male mice (18-20 g) were administered APAP by gavage after eight weeks of a high-fat diet. Any resulting vascular oxidative stress and inflammation were assessed. Levels of Ang Ⅱ, CAV1, and other related proteins were measured using ELISA and western blotting. In APAP-treated NAFLD mice, CAV1 expression was downregulated and Ang Ⅱ expression was upregulated compared to normal APAP-treated mice. In vitro, HUVECs were incubated with Ang Ⅱ (300 nM) for 48 h. Overexpression of CAV1 in HUVECs attenuated Ang Ⅱ-induced oxidative stress and inflammation and downregulated the expression of Protein kinase C (PKC) and p-P38/P38. After intervention with CAV1-siRNA, immunofluorescence results showed that the fluorescence intensity of PKC on mitochondria was further increased, and flow cytometry results showed that the mitochondrial membrane potential increased. PKC inhibitors alleviated Ang Ⅱ-induced endothelial injury. In conclusion, our findings confirmed that CAV1 exerts a protective effect against vascular injury by inhibiting oxidative stress and inflammation through the PKC/MAPK pathway. Therefore, restoration of CAV1 may have clinical benefits in reducing APAP-induced vascular damage in NAFLD patients.
对乙酰氨基酚(APAP)是世界上使用最广泛的药物之一。文献表明,过量或长期使用APAP会导致心血管功能障碍增加。在脂肪性肝病中,APAP使用引起的血管紧张素Ⅱ(AngⅡ)急性增加可能会增加血管损伤的风险和严重程度。然而,其潜在机制仍不清楚。小窝蛋白-1(CAV1)是一种广谱激酶抑制剂,对内皮功能有重要决定作用。本研究旨在观察APAP对非酒精性脂肪性肝病(NAFLD)血管系统的影响,并确定CAV1是否能通过靶向AngⅡ或其下游途径减轻血管氧化应激和炎症。在本研究中,7周龄的C57BL/6雄性小鼠(18 - 20克)在高脂饮食8周后通过灌胃给予APAP。评估由此产生的血管氧化应激和炎症。使用酶联免疫吸附测定(ELISA)和蛋白质印迹法检测AngⅡ、CAV1和其他相关蛋白的水平。与正常APAP处理的小鼠相比,APAP处理的NAFLD小鼠中,CAV1表达下调,AngⅡ表达上调。在体外,人脐静脉内皮细胞(HUVECs)与AngⅡ(300 nM)孵育48小时。HUVECs中CAV1的过表达减轻了AngⅡ诱导的氧化应激和炎症,并下调了蛋白激酶C(PKC)和磷酸化P38/总P38的表达。用CAV1小干扰RNA(CAV1-siRNA)干预后,免疫荧光结果显示线粒体上PKC的荧光强度进一步增加,流式细胞术结果显示线粒体膜电位增加。PKC抑制剂减轻了AngⅡ诱导的内皮损伤。总之,我们的研究结果证实,CAV1通过PKC/丝裂原活化蛋白激酶(MAPK)途径抑制氧化应激和炎症,对血管损伤发挥保护作用。因此,恢复CAV1可能对减少NAFLD患者中APAP诱导的血管损伤具有临床益处。