Yang Jian, Zhai Yuhong, Huang Cuiyuan, Xiang Zujin, Liu Haiyin, Wu Jingyi, Huang Yifan, Liu Li, Li Wenqiang, Wang Wei, Yang Jun, Zhang Jing
Department of Cardiology, The First College of Clinical Medical Science, China Three Gorges University, Yiling Road 183, Yichang, 443000, Hubei, China.
Hubei Key Laboratory of Ischemic Cardiovascular Disease, Yichang, 443000, China.
Inflammation. 2024 Aug;47(4):1371-1385. doi: 10.1007/s10753-024-01982-y. Epub 2024 Apr 3.
Although our previous studies have established the crucial role of RP105 in myocardial ischemia/reperfusion injury (MI/RI), its involvement in regulating oxidative stress induced by MI/RI remains unclear. To investigate this, we conducted experiments using a rat model of ischemia/reperfusion (I/R) injury. Adenovirus carrying RP105 was injected apically at multiple points, and after 72 h, the left anterior descending coronary artery was ligated for 30 min followed by 2 h of reperfusion. In vitro experiments were performed on H9C2 cells, which were transfected with recombinant adenoviral vectors for 48 h, subjected to 4 h of hypoxia, and then reoxygenated for 2 h. We measured oxidative stress markers, including superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities, as well as malondialdehyde (MDA) concentration, using a microplate reader. The fluorescence intensity of reactive oxygen species (ROS) in myocardial tissue was measured using a DHE probe. We also investigated the upstream and downstream components of the signal transducer and activator of transcription 3 (STAT3). Upregulation of RP105 increased SOD and GSH-Px activities, reduced MDA concentration, and inhibited ROS production in response to I/R injury in vivo and hypoxia reoxygenation (H/R) stimulation in vitro. The overexpression of RP105 led to a decrease in the myocardial enzyme LDH in serum and cell culture supernatant, as well as a reduction in infarct size. Additionally, left ventricular fraction (LVEF) and fractional shortening (LVFS) were improved in the RP105 overexpression group compared to the control. Upregulation of RP105 promoted the expression of Lyn and Syk and further activated STAT phosphorylation, which was blocked by PP2 (a Lyn inhibitor). Our findings suggest that RP105 can inhibit MI/RI-induced oxidative stress by activating STAT3 via the Lyn/Syk signaling pathway.
尽管我们之前的研究已经证实了RP105在心肌缺血/再灌注损伤（MI/RI）中的关键作用，但其在调节MI/RI诱导的氧化应激中的作用仍不清楚。为了研究这一点，我们使用缺血/再灌注（I/R）损伤大鼠模型进行了实验。携带RP105的腺病毒多点注射到心尖，72小时后，结扎左冠状动脉前降支30分钟，然后再灌注2小时。对H9C2细胞进行体外实验，用重组腺病毒载体转染48小时，进行4小时缺氧处理，然后再复氧2小时。我们使用酶标仪测量氧化应激标志物，包括超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GSH-Px）活性以及丙二醛（MDA）浓度。使用DHE探针测量心肌组织中活性氧（ROS）的荧光强度。我们还研究了信号转导和转录激活因子3（STAT3）的上游和下游成分。RP105的上调增加了SOD和GSH-Px活性，降低了MDA浓度，并抑制了体内I/R损伤和体外缺氧复氧（H/R）刺激引起的ROS产生。RP105的过表达导致血清和细胞培养上清液中心肌酶LDH降低，梗死面积减小。此外，与对照组相比，RP105过表达组的左心室射血分数（LVEF）和缩短分数（LVFS）得到改善。RP105的上调促进了Lyn和Syk的表达，并进一步激活了STAT磷酸化，这被PP2（一种Lyn抑制剂）阻断。我们的研究结果表明，RP105可以通过Lyn/Syk信号通路激活STAT3来抑制MI/RI诱导的氧化应激。
