丹酚酸B介导Elovl6/Echs1/Acot1通路调节脂肪酸代谢减轻H9c2细胞OGD/R损伤  

Salvianolic acid B mediates Elovl6/Echs1/Acot1 pathway to regulatefatty acid metabolism and attenuates OGD/R injury in H9c2 cells

作　　者：曹策 宋鉴书 杨丽丽[1] 李浩然 刘子馨 李磊[1] 付建华[1] 刘建勋[1] CAO Ce;SONG Jian-shu;YANG Li-li;LI Hao-ran;LIU Zi-xin;LI Lei;FU Jian-hua;LIU Jian-xun(Institute of Basic Medical Sciences of Xiyuan Hospital,Beijing Key Laboratory of Chinese Materia Pharmacology,National Clinical Research Center of Traditional Chinese Medicine for Cardiovascular Diseases,China Academy of Chinese Medical Sciences,Beijing 100091,China)

机构地区：[1]中国中医科学院西苑医院基础医学研究所,中药药理北京市重点实验室,国家中医心血管疾病临床医学研究中心,北京100091

出　　处：《中国药理学通报》2025年第3期482-490,共9页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 82030124,82174015);中国中医科学院西苑医院能力提升项目(No XYZX0303-03)。

摘　　要：目的观察丹酚酸B(salvianolic acid B,SalB)抗H9c2细胞氧糖剥夺/再复(oxygen glucose deprivation/re-oxygenation,OGD/R)损伤机制。方法CCK-8筛选SalB对OGD/R损伤H9c2细胞保护浓度;试剂盒检测SalB对乳酸脱氢酶(lactate dehydrogenase,LDH)、天冬氨酸转氨酶(aspartate transaminase,AST)、肌酸激酶(creatine kinase,CK)的影响;转录组测序探索SalB对OGD/R损伤H9c2作用机制;分子对接检测SalB和差异蛋白结合情况;ELISA法检测脂肪酸含量;RT-qPCR和Western blot验证差异基因表达水平;孟德尔随机化验证SalB作用靶点与HF因果关系。结果与模型组相比,SalB可保护OGD/R损伤后H9c2细胞,且明显降低CK、LDH和AST含量;转录组筛选差异基因100个,涉及脂肪酸延长、癌症中心碳代谢、色氨酸代谢通路;分子对接显示SalB与差异蛋白具有良好结合能;核心基因Elovl6、Echs1、Acot1 mRNA和蛋白表达与转录组一致;SalB可降低OGD/R损伤后脂肪酸含量;孟德尔随机化表明SalB可调节脂肪酸代谢,降低HF风险。 结论 SalB下调Elovl6、Echs1、Acot1表达水平,通过脂肪酸代谢通路保护OGD/R损伤后H9c2细胞。Aim To observe the mechanism of salvianolic acid B(SalB)against oxygen glucose deprivation/re-oxygenation(OGD/R)injury in H9c2 cardiomyocytes.Methods The protective concentration of SalB against OGD/R-injured H9c2 cardiomyocytes was screened by CCK-8 assay.The levels of lactate dehydrogenase(LDH),aspartate transaminase(AST)and creatine kinase(CK)were detected by ELISA kit.The mechanism of action of SalB on OGD/R-injured H9c2 cardiomyocytes was explored using high-throughput sequencing of the transcriptome.The binding of SalB to differential proteins was assessed using molecular docking assays.Fatty acid content was determined using free fatty acid kits.The relative expressions of mRNA and protein of differential genes were verified by RT-qPCR and Western blot.The causal relationship between the target of action of SalB and heart failure was examined by Mendelian randomization experiment.Results SalB protected OGD/R-injured H9c2 cardiomyocytes and significantly reduced the levels of CK,LDH and AST compared with the blank control group.One hundred differential genes were screened by transcriptome sequencing,which were mainly involved in fatty acid elongation,central carbon metabolism of cancer,tryptophan metabolism pathways.Molecular docking showed that SalB had good binding energy to differential proteins.The mRNA and protein expression of core differential genes Elovl6,Echs1 and Acot1 were consistent with the transcriptome sequencing results.SalB reduced fatty acidsafter OGD/R injury.Mendelian randomization experiments suggested that SalB might reduce the risk of heart failure through fatty acid metabolism,thereby reducing the risk of heart failure.Conclusion SalB can protect H9c2 cardiomyocytes after OGD/R injury by down-regulating Elovl6,Echs1 and Acot1 expression through the fatty acid metabolism pathway.

关 键 词：丹酚酸B 转录组高通量测序 H9C2心肌细胞 OGD/R 缺血/再灌注损伤 脂肪酸代谢 

分 类 号：R-332[医药卫生] R284.1R329.24R329.411R344.7R541.61