| Chapter 1 Effects of vitamin D supplementation on pulmonary function and quality of life in patients with chronic obstructive pulmonary diseaseObjective To investigate the effect of vitamin D supplementation on pulmonary function and quality of life in patients with chronic obstructive pulmonary disease(COPD)with severe smoking and vitamin D deficiency.Methods 60 men with severe smoking and vitamin D deficiency(serum 25 hydroxyvitamin D ≤ 20 ng/ml)were randomly divided into 2 groups.The control group(30 cases)was given routine treatment,the experimental group(30 cases)was given vitamin d3 400mg on the basis of routine treatment,oral,bid × 6 months,all patients were followed up for 6 months.Results There were no significant differences in the scores of CAT questionnaire,FEV1/FVC(%)and FEV1(%)between the 2 groups before treatment(P>0.05).After 6 months of treatment,compared with the control group,the FEV1(%)and FEV1/FVC(%)in the experimental group were significantly improved(all P<0.05),the score of CAT questionnaire was significantly reduced(P<0.05),and the number of acute exacerbations in the experimental group was significantly smaller than that in the control group after 6 months of follow-up(P<0.05).Conclusion The results showed that vitamin D supplementation can improve the lung function and quality of life in male COPD patients with severe smoking and vitamin D deficiency,and reduce the number of acute exacerbations.Chapter 2 Effects of cigarette smoke extract on macrophage autophagy and intervention effects of vitamin DSection 1 Effects of cigarette smoke extract on autophagy of RAW264.7 macrophagesObjective To investigate the effect of cigarette smoke extract(CSE)on macrophage autophagy.Methods RAW264.7 macrophages were exposed to CSE in the laboratory.Western blot was used to detect the effects of different concentrations(0.5%,1%,2%)and different time(6 h,12 h,24 h)of CSE on the microtubule associated protein 1 light chain 3(LC3)-Ⅰ and LC3-Ⅱ of RAW264.7 macrophages.Results Western blot showed that CSE could promote the expression of LC3-II and the transformation of LC3-Ⅰ to LC3-Ⅱ,and the higher the concentration,the more significant the effect was,and it was concentration dependent.Compared with the control group,CSE can promote the expression of autophagy related protein LC3-II,promote the transformation of LC3-Ⅰ to LC3-Ⅱ,and the longer the treatment time,the more significant the effect was,with time-dependent.Conclusion CSE can activate macrophage autophagy in a concentration and time dependent manner.Section 2 Intervention effect of vitamin D on autophagy of macrophages exposed to cigarette smoke extractObjective To investigate the effect of vitamin D on autophagy of macrophages exposed to CSE.Methods Different concentrations of vitamin D(0.1 nM,InM,10nM)were added to RAW264.7 macrophages exposed to 2%CSE for 24 h.Western blot was used to detect the expression of LC3-Ⅰ,LC3-Ⅱ and p62.Immunofluorescence cell assay was used to detect autophagosome and autolysosome.Results Western blot showed that compared with the control group,2%CSE could promote the expression of LC3-Ⅱ,promote the transformation of LC3-I to LC3-II,vitamin D could inhibit the expression of LC3-Ⅱ,and the higher the concentration,the more obvious the inhibition effect.Compared with the control group,2%CSE can inhibit the expression of p62,vitaminD can significantly promote the expression of p62,and the higher the concentration,the more obvious the promotion effect.Observation of LC3 spots:LC3 spots in the control group are significantly fewer,and are mainly yellow spots after colocalization,representing autophagosomes;LC3 spots are significantly increased in the 2%CSE group,and red spots are increased after colocalization,representing Autophagolysosome.The spots in the 2%CSE+10 nM VD group were significantly fewer than those in the 2%CSE group,and the spots were mainly red.Conclusion Vitamin D can inhibit CSE induced autophagy of RAW264.7 macrophages. |
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