| BackgroundThe acute lung damage (acute lung injury, ALI) is the clinical common seriously injured disease, the case fatality rate is high. In the cause of death one major part is hard-to-control the lung fibrosis. The pharmacology study indicated that ginger flavin (Curcumin) is the traditional Chinese medicine turmeric yellow display pharmacological action essential component, is one kind of relative molecular weight quite low natural polyphenols chemical compound. Not only along with many experimental studies, discovers the ginger flavin has anti-tumor, oxidation resistance, anti-SARS, anti-fibrosis and other functions. Moreover the poisonous side effect is quite low. Recent years’s research indicated that the ginger flavin can raise TGF-β1 and acceptor’s expression when ALI, and can improve the lung fibrosis, plays the protective function to ALI. The research indicated that the Smad family protein is in TGF-β1 acceptor’s most important butcher the kinase substrate, the widespread participation and have led the regulation of TGF-β1 pair of gene. But the ginger flavin prevents and controls the ALI mechanism, intervenes the progress of ALI is whether related with TGF-β1/smads signal passage, at present was still not clear.ObjectivesStudies at this experiment, within a one-time infusion of bleomycin administration trachea model, observe and detect indicators curcumin early after acute lung injury in rats. The discussion ginger flavin in the early application that rat acute lung damages the protective functions and TGF-β1/smads signal passage the effects on ALI.Methods1. Preparation of the grouping and model:Healthy male SD 36 were divided willfully became 3 groups (n=12):normal control group, model group and curcumin group. Intratracheal bleomycin using disposable infusion administration model.2.Administration and drawn:modeling the first two days of curcumin group were given 100mg·kg·-d-1 orally, model group with the same bulk of NS. Medication 3 days,7 days the rats were killed in batches. The left lobe of the lung take appropriate fixed paraffin-embedded tissue dehydration. Right lung and separated into small pieces of leaf tissue stored in EP tube, transferred to a liquid nitrogen tank spare frozen at -80℃.3.Histopathological examination:slices,fixed,conventional HE, Masson staining, microscopy.4.Detection of related proteins:The use of streptavidin-biotin-peroxidase method SABC lung smad2/3, smad7and TGF-β1immunohistochemical staining. Semi-quantitative image analysis on immunohistochemical analysis of management system of experimental results for image analysis using Image-Pro Plus6.0 image Media Cybernetics company, the lung tissue sections at each time point randomly stained area five high power field (x400), surveys and records the IOD value that each field of vision masculine gender dyes. Western blotting detection of TGF-β1, smad2/3 and smad7.5. Statistics processing:According to obtaining the data information (x±s) carries on the statistical analysis. Uses SPSS 19.0 to analyze, many sample mean value comparisons use the One-way ANOVA examination, takes P< 0.05 has the statistical significance as the difference.Results1.The general state of rats:In the experimental period, no significant changes in the control group of rats. Model rats were gradually emerge diet, decreased water intake, poor activity, occasional loose stools, lack luster fur, dry messy, lethargy, slow response, shortness of breath. Curcumin treatment of rats with the extension of time, eating and water intake were increased, the activity gradually restored, shiny fur, but a little messy, mental state also will be gradually improved, gradually smooth breathing. General state of curcumin group were better than the control group of rats, but significantly better than the model group.2. HE result:HE staining observed under the microscope:normal rats in 3d and 7d no abnormal lung tissue structure.3d model group rats acute inflammation of the lung tissue alveolar based, capillary congestion expansion, alveolar inflammatory exudate visible; when 7d some alveolar septal thickening,wall thickening, alveolar septa and alveolar inflammatory cell infiltration increased compared 3d serious.Curcumin group in 3d and 7d in the lung tissue, part bronchioles and scattered inflammatory cells, as compared with the model group relieve inflammation and injury are the alveoli.3. Masson result:Blank control group rat’s lung tissue slice may discover that only separates extremely few blue collagenous fiber in the bronchiole surroundings and pulmonary alveolus, but in the 7d model group rat dyes the blue collagenous fiber obviously in the pulmonary alveolus separates as well as around the bronchiole and minute vessel.7d ginger flavin group and 7d model group compares, all levels of bronchial tube surrounding dyeing becomes the blue collagenous fiber obvious reduction but to be higher than the blank control group.4. Immunohistochemical results:The results of the analysis show the mode group, TGF-β1,smad2/3 expression were higher than the normal group (P<0.01,P<0.01). Curcumin group TGF-β1, smad2/3 expression was decreased compared with the model group (P<0.01,P<0.01). Smad7 expression in model group was lower than that group (P<0.01,P<0.01). Smad7 curcumin increased expression compared with model group (P<0.01).5. Western blotting:the lung tissue TGF-β1 protein levels higher than the control group in the model group and curcumin group(P<0.01,P<0.01), curcumin group than the control group(P<0.01); the model group smad3 protein expression in lung tissue levels greater than the control group and curcumin group(P<0.01,P<0.05), curcumin group than the control group(P<0.01); smad7 protein expression in rat lung tissue within the model group were lower than the control group and curcumin, curcumin group than in the control group(P<0.01,P<0.01).Conclusions1.Early application of curcumin may interfere ALI evolutionary development process, and to play a protective role in acute lung injury in rats.2.Of curcumin on acute lung injury in rats of early intervention and treatment mechanism may be raised smad2/3 cut smad7 protein and protein, and thus affect the TGF-β1/smads signal pathways. |
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