The Study Of Resveratrol On Obese Asthmatic Rat Model Of Oxidative Stress Inhibition And Its Mechanism

Backgrounds:Bronchial asthma is a chronic disease with pathological changes in airway allergic inflammation,abnormal changes in airway wall structure,and increased tracheal-bronchial responsiveness.Epidemiology shows that obesity increases the prevalence and morbidity of asthma.When obese individuals are accompanied by asthma,their response to choline drugs and glucocorticoids is often poor.At the same time,obese people are more likely to have wheezing symptoms than non-obese people.Quality reduction can improve its symptoms.Oxidative stress means that the body’s highly active molecules are produced too much,and the degree of oxidation exceeds the removal of oxides.Degree,oxidation system and anti-oxidation system imbalance,resulting in tissue damage.Many systems and substances are activated during the course of bronchial asthma,including complement system,platelet-activating factor（PAF）,arachidonic acid metabolism,ROS and NO,and consequently inflammatory disorders,coagulation disorders,local Ischemia and other causes of tissue cell damage,the ultimate occurrence of organ dysfunction or even death.The Keap1-Nrf2 pathway is active in a variety of respiratory diseases,including asthma.In the absence of stress,Nrf2 interacts with its negative regulatory factor Keap1 and forms an Nrf2 / Keap1 complex.Nrf2 is degraded by the degradation of the complex protein.However,in the stress state,the P-Nrf2 / Keap1 complex is transferred to the nucleus and is combined with the antioxidant response element（ARE）to activate a variety of antioxidant genes.Keap1-Nrf2 pathway plays an important role in the biological processes such as anti-inflammatory and anti-oxidative stress.Purpose1.To establish a rat model of high fat diet obesity and asthma,to identify the common pathogenesis of obesity and asthma-oxidative stress,to provide a basis for the subsequent exploration of resveratrol by inhibiting Keap1 / Nrf2 pathway which was associated with oxidative stress.2.To further investigate the inhibitory effect of resveratrol on oxidative stress in rats by further investigating the important role of Keap1-Nrf2 pathway in the pathogenesis of obese asthmatic rat model,and further Learn the important role Keap1-Nrf2 pathway in which to play.Result The first part: the establishment of obesity,asthma animal model,grouping and related indicators of detection1.Compared with obesity,asthma,obesity + asthma group,the blood glucose level of0.03% RSV asthma group,0.03% RSV obese group,0.03% RSV obesity + asthma group was significantly lower than that of obesity,asthma and obesity group.2.The levels of high density lipoprotein cholesterol,triglyceride,total cholesterol and very low density lipoprotein were measured and compared: 0.03% RSV asthma group,0.03% RSV obese group,0.03% RSV obesity + asthma group high density lipoprotein cholesterol Significantly increased,triglyceride,total cholesterol,very low density lipoprotein was significantly lower,with a significant difference compared with obesity,asthma,obesity + asthma group.3.HE staining results showed that the adipocytes of heart,lung and kidney were clear and the structure was regular.Obesity,asthma,obesity + asthma group hypertrophy,cell number decreased,cell spacing increased,the cells arranged between the close,during which scattered in the distribution of inflammatory cells,0.03% RSV asthma group,0.03% RSV obese group,0.03% RSV Obesity + asthma group after the role of cell hypertrophy,the number increased significantly.Part 2: Inhibitory effect of resveratrol on oxidative stress in obese and asthmatic rats and its mechanism1.The levels of reactive oxygen species in the heart,lung and kidney tissues of obese,asthmatic,obese and asthmatic groups were significantly higher than those in the normal control group,0.03% RSV asthma group,0.03% RSV obese group,0.03% RSV obesity + asthma group Significantly decreased,compared with obesity,asthma,obesity+ asthma group was significantly different.2.The activities of GSH,GSH-Px,H₂O₂ and SOD in the hearts,lungs and kidneys of rats were measured.Further clarify the oxidative stress levels of obese and asthmatic animal models and the inhibitory effect of resveratrol.The results showed that the activity of oxidative stress-related enzymes in the heart,lung and kidney tissues of obese,asthmatic,obese and asthmatic groups was significantly lower than that of the normal control group,0.03% RSV asthma group,0.03% RSV obese group,0.03% RSV obesity + asthma The activity of oxidative stress-related enzymes in the group was significantly higher than that in obese,asthma,obese and asthma groups.3.Real-time RT-PCR showed that there were significant differences in the levels of Keap1 and Nrf2 at m RNA level among different groups.4.The expression of Keap1 and Nrf2 in the heart,lung and kidney cells were significantly different among differentgroups.Western blot analysis showed that the antioxidant genes Keap-1 and Nrf2 were significantly different among differentgroups.5.3T3-L1 adipocyte culture and induced differentiation: oil red staining results in the cytoplasm contains a lot of light red lipid droplets,suggesting that 3T3-L1 adipocytes induced differentiation succeeded.6.Compared with the blank control group,the levels of ROS in 3T3-L1 adipocytes group were significantly increased,the level of ROS was significantly decreased in10μmol/L RSV + 3T3-L1 adipocyte group and 50μmol/L RSV + 3T3-L1 adipocyte group,Indicating that resveratrol for 3T3-L1 adipocytes ROS levels have a significant inhibitory effect.7.The activity of oxidative stress-related enzymes in the 50μmol/L RSV + 3T3-L1 adipocyte group was significantly higher than that in the control group（P<0.05）.This indicates the inhibitory effect of resveratrol on oxidative stress in 3T3-L1 adipocytes.8.The expression level of Keap-1 m RNA in the 50μmol/L RSV + 3T3-L1 adipocyte group was significantly lower than that in the control group,10μmol/L RSV + 3T3-L1 adipocyte group,50μmol/L RSV + 3T3-L1 adipose tissue group Nrf2 m RNA expression levels were significantly increased compared with the control group was significantly different.Conclusion1.This study successfully established animal models of asthma rats with a high-fat diet obesity.2.It clear the oxidative stress playing an important role in the development of obesity and the occurrence of asthma.3.It is confirmed that Keap-1 / Nrf2 signaling pathway in asthma obesity play an important role in the oxidative stress reaction.4.It is found that the resveratrol can activate the Keap1 / Nrf2 signal regulating circuit,increase antioxidant molecules GSH release,increase the activity of antioxidant enzymes,such as gsh-px,H₂O₂ enzyme and enzyme SOD.Redusing the risk of obesity and asthma in mice caused by inflammation of reactive oxygen species level.The results provide a new train of thought for asthma and asthma diagnosis and treatment of obesity.