脑啡肽和褪黑素对失血性休克心肌细胞RIPK3表达的影响
| 作者: |
1高雅,
1刘伟,
1李璐,
1董敬之,
1刘睿,
1彭细娟,
1姚立农
1 空军军医大学第二附属医院麻醉ICU，西安 710038 |
| 通讯: |
姚立农
Email: yaolin@fmmu.edu.cn |
| DOI: | 10.3978/j.issn.2095-6959.2022.01.004 |
| 基金: | 空军军医大学第二附属医院科技创新发展基金(2018JSYJ003)；空军军医大学校发展基金(2020XB042)。 |
摘要
目的：观察失血性休克时脑啡肽(D-Ala2-Leu5-enkephalin，DADLE)和褪黑素对受体相互作用蛋白3(receptor-interacting protein kinase 3，RIPK3)、三磷酸肌醇受体/电压依赖性阴离子通道(1,4,5-trisphosphate receptor/voltage dependent anion channels，IP3R/VDAC)及氧自由基(reactive oxygen species，ROS)表达的影响，探讨其心肌保护作用的机制。方法：将30只雄性SD大鼠随机分为5组，分别为Shock组、NaCl组、Melatonin组、DADLE组、DM组，大鼠休克后分组复苏，复苏至MAP为55~60 mmHg并维持3 h。采用蛋白质印迹法检测心肌组织RIPK3及黄嘌呤氧化酶(xanthine oxidase，XO)表达水平；免疫组织化学观察心肌组织IP3R及VDAC表达水平；免疫荧光检测心肌ROS表达水平。结果：与Melatonin组、DADLE组相比，DM组RIPK3表达水平虽然无统计学意义(P>0.05)，但DM组RIPK3表达水平整体降低；与Melatonin组、DADLE组相比，DM组XO表达水平显著降低(P<0.05)。DM组IP3R表达水平与Melatonin组、DADLE组相比整体降低。DM组与Melatonin组、DADLE组相比，DM组VDAC表达水平显著降低(P<0.05)。ROS表达水平整体较其他组降低，DM组与Melatonin组、DADLE组相比差异无统计学意义(P>0.05)。结论：失血性休克早期复苏时应用DADLE、褪黑素可降低RIPK3及IP3R/VDAC表达，通过抑制XO减少ROS生成。
关键词：
失血性休克；脑啡肽；褪黑素；心肌细胞；受体相互作用蛋白3
Effects of DADLE and melatonin on expression of RIPK3 in hemorrhagic shock cardiomyocytes
CorrespondingAuthor: YAO Linong Email: yaolin@fmmu.edu.cn
DOI: 10.3978/j.issn.2095-6959.2022.01.004
Foundation: This work was supported by the Science and Technology Innovation Development Fund of the Second Affiliated Hospital of PLA Air Force Military Medical University (2018JSYJ003) and the PLA Air Force Medical University Development Fund (2020XB042), China.
Abstract
Objective: This study intends to observe the effects of D-Ala2-Leu5-enkephalin (DADLE) and melatonin on the expression of receptor-interacting protein kinase 3 (RIPK3), 1,4,5-trisphosphate receptor/voltage dependent anion channels (IP3R/VDAC) and reactive oxygen species (ROS) during hemorrhagic shock and to explore the mechanism of their protective effects on myocardium. Methods: Thirty male SD rats were randomly divided into a Shock group, a NaCl group, a Melatonin group, a DADLE group and a DM group. The rats were resuscitated after shock and were resuscitated to 55–60 mmHg for 3 h. The expression level of RIPK3 and xanthine oxidase (XO) in myocardium was detected by Western blotting, the expression level of IP3R and VDAC in myocardium was observed by immunohistochemistry, and the expression level of ROS in myocardium was detected by immunofluorescence. Results: Compared with Melatonin group and DADLE group, the expression level of RIPK3 in DM group was not statistically significant (P>0.05). However, the expression level of RIPK3 in DM group was significantly lower than that in Melatonin group and DADLE group, and the expression level of XO in DM group was significantly lower than that in Melatonin group and DADLE group (P<0.05). The expression level of IP3R in DM group was lower than that in Melatonin group and DADLE group as a whole. The expression of VDAC in DM group was significantly lower than that in Melatonin group and DADLE group (P<0.05). The expression level of ROS in DM group was lower than that in other groups. However, there was no significant difference between DM group, Melatonin group and DADLE group (P>0.05). Conclusion: DADLE and melatonin can reduce the expression of RIPK3 and IP3R/VDAC during early resuscitation of hemorrhagic shock, and reduce the production of ROS by inhibiting XO.
Keywords：
hemorrhagic shock; D-Ala2-Leu5-enkephalin; melatonin; cardiomyocytes; RIPK3