G Protein-coupled Estrogen Receptor Attenuation Of Ang ? Induced Cardiac Hypertrophy And Inhibition Autophagy Via PI3k-Akt-mTOR Signaling

Objective:To investigate the effects of GPER1(G protein-coupled estrogen receptor 1)on the hypertrophy of Ang ? induced cardiomyocytes,bioinformatics analysis was used to screen possible key proteins and the possible regulatory mechanisms were explored by cell experimental methods.Methods:1.Rat myocardial cells were cultured in vitro.The morphology of the cells was observed under a microscope and measured.The expression of GPER1 in cardiomyocytes was detected by immunofluorescence.2.Using TMT protein mass spectrometry to detect the differences in protein expression between the control group,Ang ? group and Ang ? + G1 group.Pathway analysis and Go analysis to screen for possible regulatory mechanisms.3.According to the experimental requirements divided into five groups:the blank control group,Ang ? group,Ang ?+G1 group,Ang ?+G1+G15 group and Ang II+G1+MK2206 group.Western-blot was used to detect the expression of GPER1 and the expression of Akt,70S6K1,4EBP1,mTOR and autophagy-associated LC3?and LC3I.4.The expression of atrial natriuretic peptide(ANP)and B-type natriuretic peptide(BNP)were detected by reverse transcription polymerase chain reaction(RTq-PCR).Results:1.Ang ? induced cardiomyocyte hypertrophy in a concentration-dependent manner with cardiomyocyte surface area and the transcription level of ANP and BNP increased.2.Cell immunofluorescence showed GPER1 expression on cardiomyocytes by under optical microscope.3.Bioinformatics analysis showed that RAP1 protein is a key protein that regulates cardiac hypertrophy.4.Western-blot results showed that the expression of p-Akt,p-70S6K1 and p-mTOR was increased in Ang?+G1 group compared with Ang? group,and the protein expression of p-4EBP1 was decreased(P<0.05).In the case of the combination antagonist G15 and the Akt blocker MK2206,this phosphorylation tendency was reversed(P<0.05).5.RTq-PCR results showed that GPER1-specific receptor activator G1 attenuated the transcription of ANP and BNP in cardiomyocytes in a concentration-dependent manner(P<0.05).This effect could be reversed by G15 and MK2206(P<0.05).6.Autophagy analysis showed that the ratio of LC3?/LC3? was significantly higher in the Ang? group than in the control group,while the ratio of LC3?/LC3? in the Ang?+G1 group was significantly lower(P<0.05).The ratio of LC3?/LC3? in Ang?+G 1+G15 group,Ang?+G1+Rapamycin group and Ang?+G1+MK2206 group was higher than that in Ang?+G1 group(P<0.05).Conclusion:The activation of GPER1 alleviated cardiac hypertrophy.RAPlgap protein is one of the key proteins involved in the regulation of cardiac hypertrophy by GPER1.The protective effect of GPER1 may be achieved through the PI3K-Akt-mTOR signaling pathway.GPER1 mediated autophagy may play a role in the regulation of cardiac hypertrophy.