Mechanism of resveratrol inhibiting inflammatory response in sepsis associated cardiomyopathy based on mammalian target of rapamycin-transcription factor EB autophagy signaling pathway
10.3969/j.issn.1008-9691.2024.06.012
- VernacularTitle:基于mTOR-TFEB自噬信号通路探讨白藜芦醇抑制脓毒性心肌损伤的实验研究
- Author:
Min'er CHEN
1
;
Yan XIAO
;
Hongguang DING
;
Yongli HAN
;
Juhao ZENG
;
Xinqiang LIU
Author Information
1. 广东省心血管病研究所,广东省人民医院,广东省医学科学院,广东 广州 510080
- Publication Type:Journal Article
- Keywords:
Resveratrol;
Sepsis associated cardiomyopathy;
Mammalian target of rapamycin;
Transcription factor EB
- From:
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care
2024;31(6):708-713
- CountryChina
- Language:Chinese
-
Abstract:
Objective To analyse whether Resveratrol can inhibit the inflammatory response in septic cardiomyopathy by modulating the mammalian target of rapamycin-transcription factor EB(mTOR-TFEB)signaling pathway.Methods A total of 32 clean-grade male Sprague-Dawley(SD)rats were selected and divided into sham operation group(Sham group),sepsis model group[cecal ligation and puncture(CLP)group],Resveratrol group(Res group)and mTOR inhibitor group(Torin1 group),with 8 rats in each group.Sepsis was induced by CLP.The sham group only underwent laparotomy and did not perform CLP.In the Torin1 group,20 mg·kg-1·d-1 of mTOR inhibitor Torin l injection was injected intraperitoneally 10 days before molding,once a day for 10 days.The Sham group and the CLP group were given the same amount of normal saline through the same route before molding.The Res group was injected intraperitoneally with 60 mg/kg Resveratrol injection before molding.At 6 hours after model establishment,the heart function was evaluated by echocardiography;Serum cardiac troponin I(cTnI)levels were detected by enzyme linked immunosorbent assay(ELISA).The mRNA expressions of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)were detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).The protein expressions of phosphorylation-mTOR(p-mTOR),TFEB(nuclear),TFEB(total)and light chain 3-Ⅱ(LC3-Ⅱ)of myocardial tissue were detected by Western blotting.The interaction between Resveratrol and mTOR was simulated using a network pharmacology approach.Results Compared with the sham group,the left ventricular ejection fraction(LVEF)and left ventricular short-axis shortening rate(FES)were significantly reduced in the CLP group[LVEF:0.37±0.02 vs.0.69±0.01,FES(%):17.8±3.1 vs.33.9±2.8,both P<0.01],serum cTnI levels were elevated(ng/L:1 935.96±47.78 vs.87.95±7.98,P<0.01),the mRNA expression levels of IL-6 and TNF-α were significantly up-regulated[IL-6 mRNA(2-ΔΔCt):26.10±2.08 vs.1.61±0.03,TNF-α mRNA(2-ΔΔCt):12.80±1.51 vs.1.26±0.22,both P<0.01),serum levels of inflammatory cytokines IL-6 and TNF-α mRNA were increased[IL-6 mRNA(2-ΔΔCt):26.10±2.08 vs.1.61±0.03,TNF-α mRNA(2-ΔΔCt):12.80±1.51 vs.1.26±0.22,both P<0.01),the expression level of p-mTOR protein in myocardial tissue was significantly increased(p-mTOR/β-actin:0.60±0.07 vs.0.30±0.05),while the protein levels of TFEB(total),TFEB(nuclear)and LC3-Ⅱwere significantly decreased[TFEB(total)/β-actin:0.52±0.08 vs.0.80±0.09,TFEB(nuclear)/H3:0.36±0.06 vs.0.09±0.07,LC3-Ⅱ/β-actin:0.25±0.08 vs.0.48±0.08,all P<0.01];Compared with the CLP group,the Res group and the Torin1 group had significantly higher LVEF and FES[LVEF:0.66±0.02,0.67±0.03 vs.0.37±0.02;FES(%):32.5±3.5,33.7±3.3 vs.17.8±3.1,both P<0.01],serum cTnI level was decreased(ng/L:1 216.88±36.66,1 225.78±35.64 vs.1 935.96±47.78,both P<0.01),mRNA levels of serum inflammatory cytokines IL-6 and TNF-α were decreased[IL-6 mRNA(2-ΔΔCt):3.91±0.46,4.14±0.39 vs.26.1±2.08,TNF-α mRNA(2-ΔΔCt):4.67±1.16,5.16±1.25 vs.12.8±1.51,both P<0.01],the p-mTOR protein expression in myocardial tissue was significantly decreased(p-mTOR/β-actin:0.22±0.05,0.24±0.06 vs.0.60±0.07,all P<0.01),the protein levels of TFEB(total),TFEB(nuclear),LC3-Ⅱwere significantly increased[TFEB(total)/β-actin:0.86±0.06,0.84±0.07 vs.0.52±0.08;TFEB(nuclear)/H3:0.96±0.08,0.86±0.07 vs.0.36±0.06;LC3-Ⅱ/β-actin:0.57±0.07,0.55±0.06 vs.0.25±0.08,all P<0.01].Network pharmacology was used to verify that resveratrol binds well to mTOR.Conclusion Resveratrol enhanced cardiac autophagy via the mTOR-TFEB pathway,thereby attenuating myocardial inflammation and subsequent cardiac injury in septic rats.
