| Idiopathic pulmonary fibrosis (Idiopathic Pulmonary, Fibrosis, IPF) in recent years has become a hot topic in respiratory system disease research. IPF belongs to diffuse pulmonary inflammatory diseases, mainly involving the lung interstitial disease, the pathology is characterized by alveolar epithelial injury, formation of fibroblast foci and excessive deposition of extracellular matrix, eventually led to the abnormal remodeling of the lung tissue. About the mechanism of IPF is still not completely clear, but also the lack of specific effective drugs and methods. Traditional immunosuppressants have certain effects on the IPF, can be used to explain the formation and development of pulmonary fibrosis immune micro environment has some effect. In recent years the literature reported effective prescription to treat pulmonary fibrosis through data mining, the results showed, supplementing qi and activating blood circulation method is the consensus of many doctors. The results of correlation analysis showed that the drug, Astragalus and Angelica on medicine (that is composed of Danggui Buxue Decoction) support and confidence in the highest [1]. Clinical often use ratio of 5:1, namely the Danggui Buxue Decoction, but ancient literature also has 5:6,2:1,4:1,10:3 and other records [2]. But the combined effect of Astragalus and Angelica on microenvironment in lung tissue of IPF mice and the role of local immune restoration and the best proportion of compatibility between the two there is no data report. The formulation of the small dose of Astragalus and Angelica group 5:1, group 1:1, group 1:5. Different ratios and doses of Astragalus and Angelica on drug effect on pulmonary fibrosis induced by bleomycin in male ICR mice model, with the death rate of the mice for pharmacodynamics index, to find the effect of Astragalus and Angelica optimal compatibility proportion of drugs and dosage. This experiment is aimed to illuminate the medicine intervention effect of Astragalus and Angelica on lung tissue of local immune balance and on the molecular mechanisms of lung tissue repair, in order to find the best treatment of Astragalus and Angelica IPF compatibility mode. It not only has the certain instruction function to the clinical use of the drug, but also provide new clues to the pathogenesis of pulmonary fibrosis and treatment.The theoretical research"Attending", "lung paralysis", "pulmonary fibrosis" as key words, search Chinese Technology Journal Full-text Database (CNKI) 2000-2014 years of literature. Selected indications or function is explicitly "attending", "lung paralysis", "pulmonary fibrosis" various clinical research, experimental study on prescription 92. Experimental study of 50 articles,42 articles on clinical research, involving 132 kinds of traditional Chinese medicine, the use frequency is 862 times the total. The use frequency in 1Otimes more than 24 taste. The results of cluster analysis divided into Qi medicine, drugs of activating blood, nourishing Yin, clearing heat and relieving cough, phlegm medicine five. Association rules analysis results obtained drugs on Association Rule 13, medicine 11 pairs. Astragalus and Angelica medicine to support the highest degree, which provides a theoretical basis for experimental study of late.The experiment research1 Effect of different proportion and dosage of Astragalus and Angelica on drug on survival status of mouse IPF80 ICR mice, male, SPF grade, weighing 18-22g, were randomly divided into treatment group and normal control group, model group, and 6 different ratios and doses of Astragalus and Angelica composition,10 rats in each group. Except the normal control group, other groups by intratracheal injection of bleomycin (5mg/kg) replicated the model of pulmonary fibrosis in mice. Second day of modeling, normal control group and model group were administrated with saline,6 Chinese medicine treatment group was given different ratios and doses of Astragalus membranaceus Angelica Decoction gavage treatment. The survival of the state and statistics the death situation of mice were observed. The results showed, the survival of the state of Chinese medicine treatment group mice by different dose and ratio of Astragalus and Angelica vary. Traditional Chinese medicine first,5,6 groups of 7 day mortality was lower among the inflammatory phase, low mortality rate 1,6 group continued to twenty-eighth days.2 Effect of different proportion and dosage of Astragalus and Angelica on drug on pulmonary morphology in IPF miceThe mouse model the same way as the experimental one, in twenty-eighth days after administration, using the method of abdominal aorta depletion of the mice were sacrificed, and quickly open the chest, cut off the lower lobe of the right lung on the 4% poly Formaldehyde Solution fixed 72 hours, and then after dehydration, wax impregnated, embedding after the steps made into paraffin sections, for HE staining and Wu Zhaofa simplified Mallory’s collagen staining, to observe the changes of lung tissue morphological changes and collagen content. HE staining and Mallory’s collagen staining showed that, the model group mice alveolar structure disappeared, around the bronchus and alveolar septum can be seen a large number of inflammatory cells infiltration, hyperemia and there was obvious exudation, fibrosis.6 Chinese medicine treatment group compared to the model group were improved significantly, in which traditional Chinese medicine in the treatment of first,6 group improved, the alveolar structure has been basically restored.3 Effect of different proportion and dosage of Astragalus and Angelica on medicine on HYP content in lung tissue of IPF miceThe existence of a large number of collagen fibers in the lung tissue of IPF mice, constitute the main component of collagen fibers of collagen hydroxyproline in collagen is, in most content, use this feature, the determination of the content of hydroxyproline in lung tissue, lung collagen fibers can be mastered catabolism. This study showed that, HYP content in lung tissue of model group mice were significantly higher than the normal control (P< 0.01), indicating that the collagen content in lung tissue of rats in model group increased significantly, the abnormal collagen metabolism. Compared with the model group, the Chinese medicine group could reduce the pulmonary HYP content, HYP content in first,6 groups of mice lung tissue decreased significantly.4 different ratios and doses of Astragalus and Angelica on the level of the effect of drugs on the expression of IPF in mouse TGF-beta, IL-6, Foxp3, ROR gamma T geneThrough on the size of the dose of different proportions of Astragalus and Angelica on drug in IPF lung tissue of the mice of TGF-, IL-6, Foxp3, ror gamma tmRNA content comparison can be seen group of the traditional Chinese medicine regulating factor of Th17/ Treg cells have a role. The expression of TGF-, IL-6, Foxp3 and tmRNA in the model group was higher than that in the normal control group. The expression of Foxp3 in the lung tissues of the mice with large and low dose of Radix 5:1 was stronger and the imbalance of Th17/Treg was restored.. High and low dosage of Astragalus and Angelica 1:5 groups to reduce the effects of expression of the TGF-significantly, Danggui amount greater inhibitory effect on TGF beta is more obvious. The expression of ROR T and IL-6 in 1:1 group was significantly decreased by the large and small dose of Radix Angelica. In front of the experimental results, large dose of Astragalus and Angelica 5:1 group of each gene expression level close to the normal group, to alleviate the significant pulmonary fibrosis, its mechanism may be related to the inhibition of the expression of Th17 differentiation key gene TGF beta and IL-6, ror gamma tmRNA and Treg differentiation gene Foxp3 mRNA expression of recovery.5 Effect of different proportion and dosage of Astragalus and Angelica on drug on tissue in mice with IPF repair related gene expression levelWhen the lung tissue injury, damage to the alveolar capillary barrier, resulting in bleeding in the lung tissue, platelets are activated and release cytokines. These cytokines can be induced endothelial cells and muscle fiber cells and epithelial cells. Abnormal activation of alveolar epithelial cells to produce medium, can be induced to differentiate into fibroblast foci formation, resulting in destruction of scar and lung structure. FGF2 can promote the formation of new blood vessels, but hasten and raise mesenchymal cells, fiber cells until the wound to start process of wound healing.VEGF effect on vascular endothelial cell growth factor, with promote vascular endothelial cell division, proliferation and promoting angiogenesis and increasing vascular permeability, role in maintaining vascular integrity. The angiogenesis is the essential condition for the repair damage and the important pathological changes of pulmonary fibrosis.. Previous studies found that bronchoalveolar stem (Bascs) cells in bleomycin leads to pulmonary fibrosis in mice model number decreased significantly, showed that the occurrence and development of Bascs quantity and may function and pulmonary fibrosis related. SCF/c-kit signal is precisely the necessary condition for the survival of BASCs. This experiment through the detection of traditional Chinese medicine group of IPF model mice lung tissue VEGF, FGF2 and c-kit, Cx43, mouse collagenl, mouse collagen3mRNA content and explore its mechanism.Compared to the control group, the mRNA level of model group, the lung tissue c-kit, FGF2 and VEGF and Cx43, mouse collagenl, mouse collagen3 increased (P<0.05 or P<0.01). Expression level of Chinese medicine groups of c-kit, Cx43, mouse collagenl, mouse collagen3mRNA were significantly lower in compared with normal control group, which traditional Chinese medicine group 1 relative expression quantity and normal group, the most close to. FGF2mRNA expression, Chinese medicine first,5 group than the normal control group, the other groups were lower than the control group. VEGFmRNA expression, Chinese medicine first,3 group were higher than the normal control group, the other groups were lower than the control group, but there was no difference. Compared with model group, expression level of Chinese medicine group c-kit, FGF2 and VEGF and Cx43, mouse collagenl, mouse collagen3mRNA decreased. Based on the above results, large dose of Astragalus and Angelica 5:1 group of each gene expression levels and the normal control group closer, weaving the may regulate injury by inhibiting the expression of c-kit, FGF2 and VEGF and Cx43, mouse collagenl, mouse collagen3mRNA. Lung local blood vessel formation and group of abnormal repair analysis, and thus play a role in anti fibrosis.6 Effect of different proportion and dosage of Astragalus and Angelica on IPF concentration in mouse Th17/Treg cellsThe study of IPF has confirmed the existence of the imbalance of Th17/Treg cells. Previous studies have found that the incidence of IPF in the process of CD4+IL-17A+Thl7 cell mediated reaction is dominant, and the percentage of CD4+CD25+Foxp3+Treg cells decreased, inhibiting inflammatory response ability is low, unable to contend with the Thl7 cells, the Th17/Treg is in a state of imbalance. The study found that in a mouse model of pulmonary fibrosis in the percentage of CD4+IL-17A+Th17 cells had no significant difference compared with the normal group, while the percentage of CD4+CD25+Foxp3+Treg cells increased slightly, and the related literature research results. In addition to the traditional third groups, the treatment group the percentage. of CD4+IL-17A+Th17 cells were higher than those in the model group, but the difference was not statistically significant. The percentage of CD4+CD25+Foxp3+Treg cells in each treatment group were lower than the model group, the difference was not statistically significant. Thl7 cell and Treg cell ratio, in addition to the traditional third group was lower than that of model group, the treatment group two patients were higher than those in the model group, but the difference was not statistically significant. CD4+IL-17A+Thl7 cell mediated responses are dominant accelerates the course of IPF development is still need further study. In this experiment, a small dose of Astragalus and Angelica 1:5 group were the highest survival rate, the number of CD4+IL-17A+Th17 cells was higher than that in model group, while the percentage of CD4+CD25+Foxp3+Treg cells decreased, which may be one of the mechanisms of its anti pulmonary fibrosis and improves survival in mice, its mechanism remains to be further research. |
