Effects Of Different Concentrations Of Ozone On The Expression Of NRF2 And HIF-1α In Cases With Metabolic Syndrome

ObjectiveCardio-cerebrovascular disease(CVDS)is considered to be the leading cause of death worldwide,including atherosclerosis,heart failure,dyslipidemia,hypertension.Metabolic syndrome(MS)is a group of diseases that cover at least three cardiovascular risk factors(obesity,dyslipidemia,hyperglycemia or type 2 diabetes)and are closely related to the progression of cardiovascular and cerebrovascular diseases.Insulin resistance,oxidative stress and inflammation are considered to be the main pathological mechanisms of MS.The prevalence of cardiovascular events and the risk of death in MS patients are about 2-3 times higher than those in non-MS patients,in which insulin resistance is the central link of MS.Previous studies have shown that nuclear factor E2 related factor 2(NRF2)plays a protective role in hyperinsulinemia through antioxidant stress pathway,and then plays a role in the prevention of cardio-cerebrovascular diseases by reducing the incidence of cardiocerebrovascular risk factors.The importance of NRF2 in obesity and insulin resistance is obvious.The potential use of NRF2 activator as a therapeutic method is an area worthy of attention and exploration.Appropriate concentration of ozone can quickly dissolve in plasma,which can cause mild to moderate oxidative stress and increase the activation of NRF2 transcription factors.The domain of NRF2 is responsible for activating the transcription of antioxidant response element(ARE).After inducing ARE transcription,a class of antioxidant enzymes obtained higher concentrations in response to transient oxidative stress of O3.The antioxidants produced include,but are not limited to,superoxide dismutase(SOD),glutathione peroxidase(GPX),glutathione S-transferase(GST),catalase(CAT),heme oxygenase-1(HO-1),NADPH quinone oxidoreductase(NQO-1),heat shock protein(HSP)and phase II enzymes of drug metabolism,which are related to the pathological mechanism of MS.Appropriate concentration of ozone can quickly dissolve in plasma,which can cause mild to moderate oxidative stress and increase the activation of NRF2 transcription factors.In the metabolism of animal cells,the perception and response to oxygen is a very important process,and the steady-state imbalance of this process plays an extremely important role in the pathophysiology of many diseases,including metabolic diseases,cancer,cardio-cerebrovascular diseases and so on.This process is mainly regulated by HIF-1 α(hypoxia inducible factor-1 α)and its regulator p VHL(Von Hippel-Lindautumorsuppressorprotein).The molecular structure of HIF1 transcription factor is helix-loop-helix structure,which can activate genes that encode proteins involved in hypoxia homeostasis.In addition,it can also induce the expression of proteins that control glucose metabolism,cell proliferation and angiogenesis.Hypoxia directly or indirectly regulates several genes involved in cell differentiation,including Epo(erythropoietin),LDHA(lactate dehydrogenase-A),ET1(endothelin-1),transferrin(transferrin),transferrinreceptor(transferrin receptor),VEGF(vascular endothelial growth factor),PDGF-Beta(platelet-derived growth factor-β)and other genes that affect glycolysis.In recent years,more and more studies have found that hypoxia plays a negative key role in the occurrence and development of diabetes and obesity,especially in the environment of hyperglycemia.In this study,it was found that hyperglycemia could seriously interfere with the expression of HIF-1 α protein,which was negatively correlated with the concentration of blood glucose.Obesity can lead to hypoxia in adipose tissue and small intestine,leading to adverse metabolic effects,including insulin resistance and non-alcoholic fatty liver disease,and insulin resistance can further promote the pathological changes of obesity.The purpose of this study was to observe the effects of different concentrations of medical ozone on the m RNA expression levels of NRF2 and HIF-1α in cases of metabolic syndrome.Methods90 cases of metabolic syndrome were selected from the Department of Neurology / Ozone Therapy Department of the Fifth affiliated Hospital of Zhengzhou University from September 2018 to September 2020 and randomly divided into three groups.The three groups were treated with ozone mixture of 20ug/ml(group 1),40ug/ml(group 2)and 56 ug/ml(group 3)(ug as ozone mass unit and ml as mixed gas volume unit).The cases were treated continuously for one week,and the specific treatment methods were as follows: 200 ml venous blood was extracted with a special aseptic blood bag,fully mixed with the same volume of medical ozone in vitro and then infused into the body.The expression of NRF2 and HIF-1 α in whole blood was measured before treatment,7 days after treatment and 14 days after treatment.At the same time,the indexes of blood routine,renal function and liver function were measured.ResultIntra-group comparison1 There was no significant difference in the m RNA expression of NRF2 and HIF-1 α in whole blood among the three groups before treatment.2 Compared with that before treatment,the contents of NRF2 and HIF-1 α in whole blood of the three groups increased significantly on the 7th day(P < 0 05).3 Compared with that before treatment,the contents of NRF2 and HIF-1 α in the three groups increased significantly on the 14 th day(P < 0 05).4 Compared with the 7th day of treatment,there was no significant change in NRF2 and HIF-1 α in the three groups on the 14 th day after treatment(P >0 05).5 Through the comparison of three time points,it was found that compared with the change degree from the seventh day to the seventh day of treatment,the change trend of the two indexes from the seventh day to the 14 th day of treatment was weaker.Comparison between groupsThe contents of NRF2 and HIF-1 α in group 2 and 3 were higher than those in group 1(P < 0 05),but there was no significant difference in the contents of NRF2 and HIF-1 α between group 2 and group 3(P > 0 05).Conclusions1 Ozone therapy can up-regulate the m RNA expression of NRF2 and HIF-1 α in the whole blood of patients with metabolic syndrome in a dose-dependent manner.2 When the therapeutic dose is above 40ug/ml,the effect of dose dependence is no longer obvious.