CTRP3 alleviates cardiac ischemia/reperfusion injury via LAMP1/JIP2/JNK signaling pathway.
CTRP3 alleviates cardiac ischemia/reperfusion injury via LAMP1/JIP2/JNK signaling pathway.
复制标题
CTRP3通过LAMP1/JIP2/JNK信号通路减轻心脏缺血/再灌注损伤
DOI：
10.18632/aging.203876
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发表时间：
2022-02-03
期刊：
影响因子：
--
通讯作者：
Chen J
中科院分区：
文献类型：
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作者：
Song Y;Zhang Y;Wan Z;Pan J;Gao F;Li F;Zhou J;Chen J
背景：C1 q/肿瘤坏死因子相关蛋白3（CTRP 3）已被报道是心肌梗死的重要调节因子。然而，CTRP 3在缺血/再灌注（I/R）损伤中的潜在分子机制仍不清楚。方法：采用大鼠心肌细胞H9 C2缺氧缺糖/复氧（OGD/R）建立心肌I/R损伤模型。在OGD/R处理的H9 C2细胞中检测CTRP 3和溶酶体相关膜蛋白1（lysosomal-associated membrane protein 1，LAMP 1）的表达。将CTRP 3（pcDNA-CTRP 3）和LAMP 1（pcDNA-LAMP 1）过表达质粒，或CTRP 3小干扰RNA（si-CTRP 3）和/或pcDNA-LAMP 1转染H9 C2细胞，检测细胞增殖、凋亡和氧化应激。进行Co-IP测定以验证CTRP 3、LAMP 1和JIP 2之间的关系。Western blot检测CTRP 3和LAMP 1在JIP 2/JNK信号通路中的作用。此外，在体内心肌I/R损伤模型的建立，以探讨CTRP 3的作用。结果如下：过表达CTRP 3和LAMP 1均能显著促进细胞增殖，抑制细胞凋亡，抑制活性氧（ROS）、丙二醛（MAD）和心肌肌钙蛋白（cTn-I）的产生，而沉默CTRP 3则相反，过表达LAMP 1可逆转CTRP 3的上述作用。CTRP 3与LAMP 1相互作用，并且CTRP 3和LAMP 1都与JIP 2结合。SP 600125（JNK抑制剂）可以恢复CTRP 3或LAMP 1过表达对JIP 2和磷酸化JNK（p-JNK）表达、增殖和凋亡的影响。此外，CTRP 3的过表达改善了体内心脏I/R损伤。结论：CTRP 3通过上调LAMP 1和激活JIP 2/JNK信号通路减轻心肌I/R损伤，可能成为I/R损伤的潜在治疗靶点。
Background: C1q/tumor necrosis factor-related protein 3 (CTRP3) has been reported to be a crucial regulator in myocardial infarction. Nevertheless, the potential molecular mechanism of CTRP3 in ischemia/reperfusion (I/R) injury remains largely unclear. Methods: The cell model of myocardial I/R injury was established by oxygen-glucose deprivation/reoxygenation (OGD/R) of rat cardiomyocyte H9C2. Expression of CTRP3 and lysosomal-associated membrane protein 1 (LAMP1) was detected in H9C2 cells treated with oxygen-glucose deprivation/reoxygenation (OGD/R). H9C2 cells were transfected with overexpression plasmids of CTRP3 (pcDNA-CTRP3) and LAMP1 (pcDNA-LAMP1), or CTRP3 small interfering RNA (si-CTRP3) or/and pcDNA-LAMP1, and cell proliferation, apoptosis and oxidative stress were testified. Co-IP assay was performed to validate the relationship among CTRP3, LAMP1 and JIP2. The role of CTRP3 and LAMP1 in JIP2/JNK pathway was evaluated with Western blot assay. Furthermore, in vivo myocardial I/R injury model was constructed to investigate the effect of CTRP3. Results: Overexpression of CTRP3 and LAMP1 both significantly promoted cell proliferation, inhibited apoptosis and the production of reactive oxygen species (ROS), malondialdehyde (MAD) and cardiac troponin (cTn-I), while silencing CTRP3 exerted the opposite effects, and LAMP1 overexpression reversed the effect of silencing CTRP3 on the aspects above. CTRP3 interacted with LAMP1, and both CTRP3 and LAMP1 bound with JIP2. SP600125 (JNK inhibitor) could restore the effects of CTRP3 or LAMP1 overexpression on the expression of JIP2 and phosphorylated-JNK (p-JNK), proliferation and apoptosis. Moreover, overexpression of CTRP3 improved cardiac I/R injury in vivo. Conclusion: CTRP3 alleviates cardiac I/R injury by elevating LAMP1 and activating JIP2/JNK signaling pathway, which may serve as a potential therapeutic target for I/R injury.