Study On The Effect And Mechanism Of Kaempferol Against High Altitude Pulmonary Edema Based On Network Pharmacology

Object:Based on network pharmacology to study the targets,molecular mechanisms and pathways of kaempferol against high altitude pulmonary edema,and on the basis of the above research results,establish a rat model of acute altitude hypoxia response and give kaempferol pretreatment to experimentally study whether kaempferol can improve high altitude pulmonary edema in rats rapidly entering the high-altitude hypoxia environment,and explore its possible mechanism.Method:1.Exploring the targets and pathways of kaempferol against high altitude pulmonary edema based on network pharmacology:Targets of kaempferol and ADME data were obtained through TCMSP database,CTD database and other public databases.We obtained the main targets of high-altitude pulmonary edema through Gen Cards and Dis Ge NET databases.Venny 21.0 was used to get the intersection target.Use the String platform to analyze protein interactions and construct a PPI network.The Metascape platform was used to analyze the"ingredient-target"network and its participating biological processes and pathways,and then Cytoscape 3.8.0 software was used to construct the"kaempferol-targets-pathways"network.2.The preventive effect and its mechanism of kaempferol on high altitude pulmonary edema:SD rats were modeled for high altitude pulmonary edema,divided into control group(normoxia+saline,group N),model group(hypoxia+saline,group M),acetazolamide control group(hypoxia+acetazolamide,group ACZ),experimental group(hypoxia+kaempferol,group KA).After 7 days of pre-administration,the acute hypoxia model was established through a low-pressure hypoxia chamber(simulating an environment of 6000 m at altitude).After modeling,the rats in each group are taken out in turn.After observing the general conditions,weighing and recording the weight,the rats were anesthetized,dissected under aseptic conditions,and plasma and lung tissue samples were collected.The pathological changes of lung tissue in each group were observed,wet/dry ratio was calculated.Superoxide dismutase activity,malondialdehyde content,nitric oxide synthase activity,the activity of Na~+-K~+-ATPase and the levels of inflammatory cytokines include tumor necrosis factor-αand interleukin-6 in plasma and lung tissue were detected.The m RNA and protein expressions of phosphatidylinositol-3-kinase and protein-serine-threonine kinase in lung.Result:1.The oral availability of kaempferol was 41.88%and the medicinal properties were 0.24.2.The main targets involved in the anti-high altitude pulmonary edema effect of kaempferol are AKT1,TNF,and IL6,and others such as BCL2,VEGFA,IL1B,MPO,and so on are also relatively important targets.The main KEGG pathways involved are the HIF-1signaling pathway and the PI3K-Akt signaling pathway.3.Animal experiment results:(1)Compared with the control group,the wet/dry ratio of the model group was significantly increased(P<0.01),the pathological manifestations of pulmonary edema were significant,the levels of TNF-αand IL-6 in lung tissue were significantly increased(P<0.05),and SOD activity was significantly decreased(P<0.05),the activity of Na~+-K~+-ATPase decreased(P<0.05).(2)Compared with the model group,the wet/dry ratio of both the acetazolamide group and the kaempferol group were reduced(P<0.01),the pathological manifestations of pulmonary edema were alleviated,the levels of TNF-αand IL-6 were reduced(P<0.05),and SOD activity significantly increased(P<0.01),NOS activity increased(P<0.05),MDA content decreased(P<0.01),the activity of Na~+-K~+-ATPase increased(P<0.05).(3)Compared with the control group,the SOD activity in plasma of model group was decreased(P<0.05),and the MDA content was significantly increased(P<0.01).Compared with model group,SOD activity of kaempferol group and acetazolamide group was slightly increased(P<0.05),MDA content was decreased compared with model group(P<0.05),and there was no significant difference between acetazolamide group and kaempferol group(P>0.05).(4)Compared with the control group,the expression of PI3K protein in the model group increased significantly(P<0.05),and the expression of AKT1 protein increased(P<0.05).The expression of PI3K in the kaempferol group was reduced comparing with the model group(P<0.01),while the expression of AKT1 protein was significantly reduced(P<0.05).(5)Compared with the control group,the PI3K m RNA content and the AKT1 m RNA content of the model group was significantly increased(P<0.01).PI3K m RNA in the kaempferol group was decreased(P<0.01)compared to the model group,and the expression of AKT1 m RNA was decreased(P<0.05).Conclusion:1.Kaempferol has good oral availability and drug-like properties,and is an active pharmaceutical ingredient with development potential.2.The targets involved in the anti-high-altitude pulmonary edema effect of kaempferol are mainly concentrated in AKT1,TNF,IL6,and the others such as BCL2,VEGFA,IL1B,MPO,etc.are also relatively important targets.Kaempferol plays a role in the prevention and treatment of high-altitude pulmonary edema through HIF-1 signaling pathway and PI3K-Akt signaling pathway.3.Kaempferol can prevent the occurrence of high-altitude pulmonary edema induced by low pressure and hypoxia by reducing oxidative damage and inhibiting inflammation.4.Kaempferol down-regulates the PI3K-Akt signaling pathway,inhibits AKT phosphory-lation,reduces oxidative stress and inflammation,thereby reducing the occurrence of high-altitude pulmonary edema.