| Objective and background:Pulmonary fibrosis (PF) is one of the most serious diseases of respiratory system.lt main show is clinically wheeze and shortness of breath, cough, cough up phlegm to restrictive ventilatory dysfunction, hypoxemia, eventually leading to respiratory failure. Epithelial mesenchymal transition(EMT) theory is one of the main achievements in recent years about international research organ fibrosis. So this topic proposed on the basis of previous research results, the clear lung collaterals treatment, selection (lung yu, kao huang yu) moxibustion, observed by bleomycin copy classical pulmonary fibrosis in rats model of epithelial cells in moxibustion therapy treatment protein. E-cadherin (E-cad) α-smooth muscle actin(α-SMA), vimentin to express the dynamic change of preliminary clarify clear lung collaterals method EMT key sites and regulation law regulation of pulmonary fibrosis.Method:8weeks health SPF rats 81, weight 180 ± 20 g, male and female half. Rats were randomly grouped, ck (C) group and 9; Model (group B):7 days (B7) group and 9,11 days (B11) group and 9,23 days(B23) group and 9,35days (B35) group and 9; Moxibustion group (M):moxibustion group 11 days (Mn) 9, moxibustion 23 days (M23) group 9, moxibustion 35 days (M35) group and 9; Positive control group (prednisone, P) 9 only. Building endotracheal one-time injection by adopting the method of Thrall to duplicate the pulmonary fibrosis model induced by bleomycin, blank control group by the method of building same endotracheal one-time injection volume of physiological saline, etc. Each building began treatment and intervention in the next day, alternating moxibustion group bilateral lung yu, paste; Prednisone prednisone group to fill the stomach; All the blank group and model group and moxibustion group to adopt the same grab is fixed, not given any treatment. Model group 7,11,23,35 days, choose the corresponding time point group rats respectively; Moxibustion group 11,23,35 days (one day) after each procedure, choose corresponding time points respectively in rats; Prednisone group, blank group selection in 35 days after all. Groups according to body weight, adopts the femoral artery bleeding to death in the rat, open the chest, observe lung in color, size, presence of bleeder, nodules, etc. Take the left lung group of formaldehyde fixed in 10% neutral, then select the left upper lobe fixation, embedding, sectioning lines after HE and Masson staining, in accordance with the classification score Szapie method alveolitis and pulmonary fibrosis grading scores. Take the organization of the upper lobe for real-time fluorescence quantitative polymerase chain reaction (QPCR) to detect E-cadherin mRNA, alpha SMA mRNA, vimentin mRNA expression. The data from the experimental process for the statistical analysis, adopt SPSS 19.0 all measurement data with standard deviation, mean soil using multi-factor analysis of variance to statistical analysis, P<0.05 showed statistically significant difference.Result:1.Each group generally observed in ratsBlank control group rats is steady breathing, hair moist bright and responsive, stool forming, normal weight growth; Model group rats shortness of breath, chest and increase faster, the nose and green purple was at onyx spiritual malaise, curled up, outlining the hair sparse lacklustre, defecate syrup, anus department or fecal sewage, weight decreased obviously; After treatment, moxibustion group compared with model group generally has a greater improvement; No difference between the positive control group with moxibustion group.2.Lung coefficientModel group, the moxibustion group and prednisone group each lung coefficient were significantly greater than the blank control group (P<0.01); Model group and moxibustion group 11 d no statistical difference (P>0.05) in model group 23 d lung coefficient of 23 d in the moxibustion group, model group,35 d lung coefficient is 35 d moxibustion group were higher, there are significant differences (P<0.01); Model group 35 d lung coefficient is high, in the positive control group was statistically difference (P<0.05); Moxibustion 35 d group and positive control group lung coefficient of no statistical significance (P>0.05).3.The pathology observation:3.1 model group:the model group 7,11 d alveolitis degree compared with blank control group were significantly increased (P<0.01),7 d degree of pulmonary fibrosis model group and blank control group, no significant difference between statistical difference (P>0.05), 11d pulmonary fibrosis model group was obviously serious in the blank control group (P<0.01); Model group,35 d alveolitis can alleviate 23 but still serious in the blank control group (P<0.05),23,35 d degree of pulmonary fibrosis model group compared with blank control group, there are significant differences (P<0.01).3.2Moxibustion group:moxibustion group 11 d alveolitis degree seriously in the blank control group (P<0.01),23 with moxibustion group,35 d reduced alveolitis degree, compared with blank control group there are still differences (P<0.05); Moxibustion group compared with model group, 11d alveolar inflammation mitigated, statistically significant (P<0.05), model group and moxibustion group 23 d has no obvious difference (P>0.05), moxibustion 35 d a model significantly reduce 35 d alveolitis have significant difference (P<0.01); Moxibustion group 11,23,35 d pulmonary fibrosis degree seriously in the blank control group(P<0.01),11 moxibustion group,23 d respectively with model group,11,23 d has no obvious difference (P>0.05), moxibustion group 35 d a model 35 d reduce pulmonary fibrosis degree, there are significant differences (P<0.01).3.3positive control group:positive control alveolitis and pulmonary fibrosis group compared with blank control group, there are significant differences (P<0.01), positive control group degree of alveolar inflammation with the model group 35 d,35 d moxibustion group were no statistical difference, (P>0.05); Positive control group compared with model group degree of fibrosis reduced 35 d, statistically significant (P<0.05), positive control group and moxibustion group 35 d fibrosis degree has no obvious difference, no statistical significance (P>0.05).4.Each group of pulmonary fibrosis protein mRNA expression4. 1E-cad mRNA expression in the pulmonary fibrosisCompared with the blank control group, model group, the moxibustion group, positive control group E-amount of mRNA expression of cad all have varying degrees of decline, in model group 35 d the lowest; Moxibustion therapy intervention after moxibustion group E-cad mRNA expression of the decline in model group is slow obviously than the same period, especially compared to the model group and 35 d, moxibustion group 35 d E-cad mRNA expression quantity increased obviously, there were significant differences (P<0.01); And positive control group compared with model group 35 d, positive control group E-cad mRNA expression increased, with statistical difference (P<0.05);4.2 α-SAM mRNA expression in the pulmonary fibrosisCompared with the blank control group, model group, the moxibustion group, positive control group a-SMA m RNA amount of expression to have the varying degree rise,35 d highest in model group; Moxibustion moxibustion group after treatment intervention on a-SMA mRNA expression of its upward trend model group is slow obviously than the same period,11th day compared with model group, moxibustion group 11 d alpha SAM mRNA expression quantity decreased significantly, there were significant differences (P<0.01); 35 days compared with model group, moxibustion group 35 d alpha SAM mRNA expression quantity has decreased, there are differences (P< 0.05). Positive control group and model group 35 d,35 d moxibustion group were no significant difference, statistically insignificant (P>0.05).4.3vimentin mRNA expression in the pulmonary fibrosisCompared with the blank control group, model group, the moxibustion group, positive control group of vimentin mRNA expression quantity rising to some extent, 35 d highest in model group; Moxibustion moxibustion group after treatment intervention on vimentin mRNA expression of its upward trend model group is slow obviously than the same period,23 days compared with model group, moxibustion group 23 d vimentin mRNA expression quantity decreased significantly, there were significant differences (P<0.01); 35 days compared with model group, moxibustion group 35 d vimentin mRNA expression quantity has decreased, there are differences (P<0.01); Positive control group and model group 35 d,35 d moxibustion group were no significant difference, statistically insignificant (P> 0.05).Conclusion:(1)the experiment by tracheal perfusion lai once obtained pulmonary fibrosis in rats model and its pathological morphological changes each time point with the domestic and foreign literatures, this experiment by bleomycin copy of classical pulmonary fibrosis in rats model was a success.(2)moxibustion on pulmonary fibrosis development stages lung tissue pathology morphology could improve the (alveolitis and pulmonary fibrosis), shows that moxibustion for prevention and control of pulmonary fibrosis have certain effect, its curative effect is related to the length of the moxibustion treatment, the longer the course of the treatment effect is better.(3)moxibustion for the treatment of pulmonary fibrosis in rats model of intervention that may be related to inhibition of EMT stromal cells among the key factors of alpha SMA, the expression of vimentin protein, increased the epithelial cells key factor E-cad protein expression level, therefore the EMT development process.(4)Vibration shenshu point of the lung, lung qi, regulating lung function; Kao huang yu, disease as virtual viscera is deficient, tonifying qi, regulating channels of moxibustion and the caves, achieved through in repair, not imaginary, no evil, peace, Yin and Yang chang qi and blood, the theory of viscera function recovery. The experiments demonstrated that using "fill" and "tong and supplement treatment to dredge lung collaterals, moxibustion for prevention and treatment of pulmonary fibrosis provides experimental theory basis. |
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