| Objective: Acute mountain sickness(AMS)is one of common diseases in plateau area,affecting the normal activities of people entering into plateau and impacting the combat effectiveness of soldiers.Heart,lung and brain are more affected in AMS.The pathogenesis of myocardial injury in AMS remain to be studied.At the same time,it is important to develop a new drug against AMS,with a serious threat to the health of people exposing to high altitude.In our study,we explore the pathogenesis of myocardial injury in AMS,the mechanism of Compound Danshen Dropping pills(CDDP)to AMS and the activity of “GY-1”anainst AMS.Methods:(1)Establish a rats model of myocardial injury in AMS.The hypoxia chamber was used to established rats model of myocardial injury in AMS,simulating hypoxic environment of 7,000 m altitude.The concentration of biochemical index in rats serum were measured by colorimetry and spectrophotometry analytical methods.The changes of myocardium in sub-cell level were detected by transmission electron microscope(TEM).Hematoxylin-Eosin(HE)staining was used to observe the histopathological changes of the myocardial.(2)The pathogenesis of myocardial injury in AMSThe changes of genes in transcription level were observed based on RNA-Seq method;The cell hypoxia model was established by low-oxygen incubator.The ROS level of mitochondrion was observed by staining with Mito Tracker Red CXMRos and laser confocal scanning microscopy.The concentration of biochemical index in culture medium supernatant were measured by colorimetry and spectrophotometry analytical methods.The ATP content of cardiomyocyte was measured by ATP Determination Kit.(3)The mechanism of CDDP against AMSThe blood oxygen saturation,the number of red blood cells and hemoglobin was were detected by oximeter and Automatic Blood Cell Analyzer.The concentration of SOD,MDA were detected using Total superoxide Dismutase Assay Kit and Lipid Peroxidation MDA Assay Kit.HE staining was used to observe the histopathological changes of the myocardial.Network pharmacology and molecular docking methods were applied to investigate the effective active ingredients and molecular mechanisms of CDDP against AMS and verify the combination of active components and corresponding targets.(4)The activity of “GY-1” anainst AMSThe blood oxygen saturation was detected by oximeter and the concentration of biochemical index in rats serum were measured by colorimetry and spectrophotometry analytical methods.Results:(1)The level of c Tn-I,CKMB,AST,LDH in HH-3d group was significantly increased.On the sub-cellular level,the mitochondrion were seen swollen with ruptured membrane,and the lipid peroxidation products of mitochondrion were seen.The Z line disappears.There was inflammatory cell infiltration in HH-3d group.(2)Compared with control group,524 significant DEGs were found,including 138 downregulated and 386 upregulated genes in HH-3d group.Our work revealed that ML351,a ALOX15 inhibitor protected myocardial cells from reducing the level of myocardial enzymes and mitochondrial ROS,increasing ATP content,inhibiting the protein expression of p-AMPK / AMPK and increasing the expression of HO-1.The therapeutic effects can further enhanced by the combined administration of ML351 and Compound C.(3)The level of lipid peroxide in myocardial tissue was decreased and the protein expression of DJ-1was increased by CDDP.The results of net pharmacology showed that compounds including quercetin,tanshinone a,ginsenoside Rh2 and genes including VEGF、 SCN5A、PTGS2 were related in AMS.(4)“GY-1” can increase blood oxygen saturation and the number of erythrocyte,hemoglobin and inhibit the release of inflammatory factors in brain.Concludes:(1)ALOX15-AMPK signal axis play important role in the pathogenesis of myocardial injury in AMS.(2)Quercetin,tanshinone a,ginsenoside Rh2 and VEGF,SCN5 A,PTGS2 were related in CDDP against AMS.(3)“GY-1”can be a candidate against AMS for further study. |
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