Study On The Effect Of Ⅱ On Tobacco Smoke Exposure - Induced Lung Injury In Mice And Its Mechanism

Objective:To observe the effect of Xiwang II on blood biochemical index and pulmonary histopathology of tobacco smoke exposure induced lung injury in mice. To study pulmonary protein expression changes of tobacco smoke exposed mice with application of proteomic technology, and investigate possible mechanism of action of Xiwang II on tobacco smoke exposure induced pulmonary injury in mice from the proteome analysis.Methods:156 male Kunming mice according to the random number table method, were randomly divided into three parts:pure tobacco smoke exposure, stop tobacco smoke exposure (ie, smoking cessation) with treatment and continued exposure to tobacco smoke with treatment, a total of 13 groups,12 mice in each group. Pure tobacco smoke exposure part is divided into ① control group:mice were exposed to the natural air for 15 days;②simple exposure model group:mice were exposed to tobacco smoke 72min daily in consistent 15 days. Exposure to tobacco smoke with treatment section is divided into ①control group:mice were exposed to the natural air for 45 days and intragastrically administrated double distilled water 5.49ml·kg-1 once daily from the 16th day; ②smoking cessation model group:mice were exposed to tobacco smoke 72min daily in consistent 15 days, then exposed to the natural air environment from the 16th day and other things were same as the control group;③ soking cessation treatment group of the procyanidin:mice were intragastrically administrated procyanidin 52mg·kg-1 once daily from the 16th day and other things were same as the smoking cessation model group;④ smoking cessation treatment groups of Xiwang II low, medium and high dosage (XwIIL, XwIIM, XwIIH):mice were intragastrically administrated Xiwang Ⅱ 1.83ml·kg-1，5.49ml·kg-1,16.47ml·kg-1 respectively once daily from the 16th day and other things were same as the smoking cessation model group;⑤ continuous exposure model group:mice were exposed to tobacco smoke 72min daily in consistent 45 days and intragastrically administrated double distilled water 5.49ml·kg-1 once daily from the 16th day;⑥ continuous exposure treatment group of procyanidin:mice were intragastrically administrated procyanidin 52mg·kg-1 once daily from the 16th day and other things were same as the continuous exposure model group; ⑦ continuous exposure treatment groups of Xiwang Ⅱ low, medium and high dosage (XwIIL, XwⅡM, XwⅡH):mice were intragastrically administrated Xiwang Ⅱ 1.83ml·kg-1,5.49ml·kg-1,16.47ml·kg-1 respectively once daily from the 16th day and other things were same as the continuous exposure model group. The levels of serum aspartatetransaminase(AST), alaninetransaminase(ALT),catalase(CAT),lactatedehydrogenase(LDH),succinate dehydrogenase(SDH),gluthathione peroxidase (GSH-PX), myeloperoxidase(MPO), malondialdehyde(MDA), total superoxide dismutase(T-SOD), Cu/Zn superoxide dismutase (Cu/Zn-SOD) were detected. T-SOD, Cu/Zn-SOD and MDA levels were detected in the upper right lobe of the liver, lung and kidney.The content of hydroxyproline(Hyp) in the lower right lobe of lung and skin were detected. The content of MPO in the upper part of the heart was detected. The left lobe of the liver, upper left lobe of lung, the heart central, between two scapula to two iliac back of skin and left kidney were fixed with 10% neutral buffered formalin for 48h and observed the morphology by HE staining, Masson staining, elastic fiber staining. Morphometry of volume ratio of hepatocytes,alveolar volume difference.ratio of alveolar cavity and the total area, mean alveolar number,collagen fiber volume ratio of lung, heart, liver and kidney, epidermal thickness, elastic fiber, expression of transforming growth factor beta 1 in mice were compared among the different groups. The mice lung protein were extracted and the expression differences of protein between the model and the treatment group and the control group were separated and identified by proteomics technology.Results:①Compared with the control group, the level of Cu/Zn-SOD, T-SOD of liver and lung tissue, CAT, SDH, GSH-PX of serum in model group decreased significantly. The level of of MDA of liver and lung tissue, hydroxyproline of lung, AST, ALT, MPO of serum in model group increased significantly.②Compared with the smoking cessation model group, the activity of T-SOD, Cu/Zn-SOD of liver tissue, GSH-PX and SDH of serum reduced significantly and AST, ALT, MPO of serum rised significantly in Xiwang Ⅱ low, medium and high dosage A groups. The activity of CAT of serum in Xiwang Ⅱ medium dosage A group rised significantly. The activity of MDA of liver and lung tissue in Xiwang Ⅱ high dosage A group reduced significantly. The activity of LDH of serum in Xiwang Ⅱ high dosage medium and high dosage A groups rised significantly. The activity of Cu/Zn-SOD of lung tissue rised significantly and MDA of lung tissue reduced significantly in Xiwang Ⅱ low, medium and high dosage A groups. The level of T-SOD of lung tissue rised significantly and hydroxyproline of lung tissue reduced significantly in Xiwang II medium and high dosage A groups.③Compared with continuous exposure model group, the level of T-SOD, Cu/Zn-SOD of liver tissue and CAT, GSH-Px, SDH of serum rised significantly; the activity of MDA of liver and lung tissue and AST, ALT, MPO of serum reduced significantly in Xiwang II low, medium and high dosage B groups. The level of hydroxyproline of lung tissue and LDH of serum reduced significantly in Xiwang II medium and high dosage B groups.④Xiwang II increased the expression of transforming growth factor beta 1in mice skin. Proteomics results show that Xiwang Ⅱ regulated the differential expression proteins were observed between the moking cessation model group and control group, continuous exposure model group and control group respectively. Compared with control group, Most of the identified differential proteins are redox enzymes, fatty acids, and involved in fatty acids, cardiac muscle contraction and other metabolic processes.Conclusion:Xiwang II applied protective effect on lung injury induced by tobacco smoke exposure and associated skin, heart, liver and kidney damage. Xiwang II may be through improving the body antioxidant content, maintenance the balance of oxidation and antioxidation, regulating lung enzyme activity, regulation of lung protein and fatty acid metabolism, so as to protect the lungs and other organs function, against oxidative damage of tobacco smoke exposure.