The Influence Of Formononetin Combined With Cimicifugin On Bleomycin Induced Pulmonary Fibrosis In Mice

The first part: Meta analysis and systematic review for the treatment of idiopathic pulmonary fibrosis with Chinese and Western MedicineObjective: To evaluate the efficacy and safety of traditional Chinese medicine combined with pirafenidone / N-acetylcysteine in the treatment of idiopathic pulmonary fibrosis(IPF).Methods: CNKI,Wan Fang data,CBM,Pub Med,EMBASE and the Cochrane library were searched by computer to find all the literatures about the treatment of idiopathic pulmonary fibrosis with traditional Chinese medicine.The deadline was set to October 31,2019.The two evaluators independently applied endnotes X7 software to screen and extract data according to the inclusion and exclusion criteria,and Revman 5.3 software to conduct quality assessment and meta analysis.Results: 10 literatures,10 in Chinese and 0 in English were included.1.Clinical comprehensive efficacy: four studies were included,and the analysis results [RR = 1.29,95% CI(1.06,1.57),P = 0.01],indicating that the comprehensive efficacy of integrated traditional Chinese and Western medicine in the treatment of idiopathic pulmonary fibrosis is better than that of Western medicine group.2.TCM syndrome score: four studies were included.The results showed that the therapeutic effect of TCM combined with TCM was better than that of Western medicine alone [SMD =-1.43,95% CI(-1.93,-0.58),P < 0.00001],the difference was statistically significant.3.Lung function: FVC%: two studies were included in the meta analysis.The diamond is located on the right side of the invalid line and does not intersect with it [SMD = 1.08,95% CI(0.28,1.89),P = 0.008],suggesting that the efficacy of pifenidone combined with traditional Chinese medicine in improving FVC% is significantly better than that of oral administration alone.FEV1%: two studies reported the effect of pifenidone combined with traditional Chinese medicine and independent medicine on FEV1%,the results were [SMD = 0.32,95% CI(-0.28,0.92),P = 0.30],suggesting that there was no significant difference between the two groups in improving FEV1%.DLCO: six studies were included for analysis.The results showed that the combination of traditional Chinese and Western medicine could improve the efficacy of DLCO [SMD = 0.36,95% CI(0.14,0.58),P = 0.001],which was significantly better than the single drug.TLC: five studies were included,and random effect model analysis was used [SMD = 0.39,95% CI(0.06,0.73),P = 0.02].The results showed that the effect of combined traditional Chinese and Western Medicine on TLC was better than that of Western medicine group.VC: five studies were included.The analysis results showed that compared with western medicine alone,the improvement of VC was not significant in combination with traditional Chinese medicine [SMD = 0.26,95% CI(-0.00,0.52),P = 0.05].The results of pulmonary function analysis showed that the combination of Chinese and Western medicine could effectively delay or improve FVC,DLCO and TLC of IPF patients compared with the single oral administration of Western Medicine(P < 0.05),but there was no significant difference between the two groups in improving FEV1 and VC.4.Arterial oxygen partial pressure(Pa O2): Meta analysis of four studies showed that [SMD = 0.62,95% CI(0.29,0.95),P = 0.005],suggesting that the combination of Chinese and Western medicine can effectively improve PaO2 compared with western medicine group,with a stable outcome.6.Dyspnea score: three studies assessed by MMRC dyspnea scale were included in the analysis,the results were [SMD =-0.41,95% CI(-0.92,-0.01),P = 0.05],suggesting that the effect of combination of traditional Chinese and Western Medicine on reducing the degree of dyspnea was not statistically significant compared with that of Western medicine group.7.Quality of life: two studies were included for meta analysis [SMD =-0.50,95% CI(-0.89,-0.12),P = 0.01],suggesting that the effect of combined medication on maintaining the quality of life of patients is better than that of Western medicine alone.The 8.6-minute walk test(6MWT)included 4 items.The results showed that the curative effect of combined medication was better than that of Western Medicine Group [SMD = 0.48,95% CI(0.14,0.82),P = 0.0002],the difference was statistically significant.8.Safety evaluation: seven studies evaluated the safety of RCT,three reported adverse reactions,and four did not show obvious discomfort 9.Other indicators: FEV1 / FVC%,SpO2,HRCT were included in the literature because of the lack of effective data,and the heterogeneity was too large.Meta analysis should be carried out after the accumulation of high-quality literature in the future.Conclusion: IPF with integrated traditional Chinese and Western medicine has a good effect in improving the comprehensive clinical effect and lung function,especially in improving the quality of life,with less adverse reactions reported.The randomized clinical control study with high quality and large sample is not enough in this study,and the results are limited.The follow-up study is still needed to provide evidence for the conclusion.The second part: The influence of formononetin combined with cimicifugin on bleomycin-induced pulmonary fibrosis in miceObjective: Yupingfeng powder is a classical prescription for the treatment of respiratory diseases.In clinical application,it is often added and reduced based on the compatibility of Astragalus and Fangfeng.The experimental study shows that the main active components of Astragalus membranaceus and Fangfeng are miscanthin and cimicin.Based on this,this study established bleomycin mice model of pulmonary fibrosis,gave miscanthin combined with cimicin intragastric intervention,observed its animal efficacy,preliminarily explored the relevant mechanisms,and provided theoretical basis for later experiment and clinical application.Methods: 42 male C57 BL / 6 mice were randomly divided into 6 groups: control group(Na Cl group),bleomycin group(BLM group),bleomycin + Menthol group(Menthol group),bleomycin + cimicin group(cimicin group),bleomycin + combination group(combination group),bleomycin + pirfenidone group(PFD group).In the Na Cl group,0.9% normal saline was infused into the trachea,and the other groups were administrated with bleomycin solution of the same dose through the trachea to establish the pulmonary fibrosis model.Seven days later,at the same time every day,the Na Cl group and BLM group were administrated with normal saline by gavage every day,and each group was administrated with 50 mg / kg of menthol,100 mg / kg of cimicin and 200 mg / kg of pirafenone respectively,and the combination group was administrated with 50 mg / kg of menthol+ Cimicin 100 mg / kg was administrated by gavage to record the change of body weight.On the 14 th day of modeling,the lung function of mice was measured to evaluate the progress of the disease and the materials were killed.The content of hydroxyproline(Hyp)in lung tissue was measured to evaluate the degree of fibrosis.He staining was used to evaluate the pathological changes of lung tissue,immunohistochemistry was used to detect the expression of COL-1 and α-SMA in lung tissue,and the molecular mechanism of anti fibrosis was explored.Results: 1.Body weight: there was no significant difference in the average body weight of mice in each group before modeling.After 7 days of modeling,except the average body weight of mice in NaCl group(g),the average body weight of mice in each group continued to drop,indicating that the modeling was successful.After 7 days of administration,it can be observed that compared with BLM group,the weight drop trend of mice in Mansi group and cimicin group was relieved,and the average body weight growth trend of PFD group and combined group was more and more obvious The potential is close to NaCl group.2.Lung function: FVC: after 7 days of administration,the FVC value of BLM group was significantly lower than that of NaCl group(P < 0.01).(2)Compared with BLM group,the FVC value of mice in awn handle group(P < 0.05)and PFD group(P < 0.01)increased,suggesting that awn handle anthocyanin and pirfenidone can effectively improve the FVC value of mice.(3)There was no significant difference between the two groups.(4)There was no significant difference between the three groups in PFD group.FEV1:(1)FEV1 in BLM group was significantly lower than that in NaCl group(P < 0.01).(2)Compared with BLM group,FEV1 of mice in awn handle group(P < 0.01),combined group(P < 0.05)and PFD group(P < 0.01)was significantly higher.(3)There was no significant difference between the two groups(P > 0.05).Cdyn:(1)the mean value of cydn in BLM group was 0.008,which was significantly lower than that in NaCl group(P < 0.01);(2)compared with BLM group,PFD group,combination group and awn handle group,they were improved by 106.1%,124.8% and 41.2% respectively,with statistical significance(P < 0.05).(3)Compared with the combined group,the cydn value of the awn handle group was significantly higher(P < 0.05),but there was no significant difference in the cimicin group.(4)The cydn of PFD group was higher than that of cimicin group(P < 0.05),and there was no significant difference between PFD group and awn handle group or combination group.Respiratory resistance(RE):(1)the mean value of re in BLM group was 1.16,significantly higher than that in Na Cl group(P < 0.01);(2)re in cimicin group was 0.84,27.4% higher than that in BLM group;re in awn handle group,PFD group and combination group were 0.76,0.72 and 0.69,respectively,significantly lower than that in BLM group(P < 0.01).(3)There was no significant difference between PFD group,awn handle group and cimicin group(P > 0.05)Inspiratory resistance(RL):(1)FVC value of BLM group was significantly lower than that of NaCl group(P < 0.01).(2)Compared with BLM group,the RL value of each group was significantly lower(P < 0.01).(3)there was no significant difference between the two groups.(4)RL value of PFD group was significantly lower than that of cimicin group(P < 0.05).To sum up,compared with BLM group,awnashi group,combination group and PFD group have some improvement on pulmonary function of bleomycin induced fibrosis mice,PFD group and awnashi group have no difference in efficacy,but cimicin group and combination group have no effect on FVC.At the same time,compared with the awn handle group and cimicin group,the combined group showed no significant advantage in improving the pulmonary function of mice with pulmonary fibrosis.3.HE staining: in the NaCl group,the alveolar structure was intact,the alveoli and pulmonary septum were basically normal,no obvious bleeding and inflammatory cell infiltration were found;in the BLM group,extensive thickening of the alveolar septum was observed,the alveolar structure was damaged,and a large number of inflammatory cells gathered and infiltrated.In PFD group and cimicin group,the infiltration area of inflammatory cells in lung tissue decreased,the degree of alveolitis and pulmonary fibrosis decreased,but there were still more fibrous tissue hyperplasia,and the degree of alveolus narrowing improved.Compared with the combined group,the inflammatory cell infiltration and alveolar structure damage in the awn handle group and BLM group were significantly relieved.4.Measurement of fibrotic area and hydroxyproline content in lung tissue: Statistics of he fibrotic area and determination of Hyp content in lung tissue(1)Hyp in BLM group was significantly higher than that in NaCl group(P < 0.05).(2)Compared with BLM group,the content of Hyp and fibrotic area of each group were significantly lower,the difference was statistically significant(P < 0.01).(3)The average Hyp and fibrosis area of cimicin group were 274.05 and 13.93%,which were significantly higher than those of combination group(P < 0.05).There was no significant difference between the two groups(P > 0.05).5.Immunohistochemical staining: the proliferation of myofibroblasts and the content of extracellular matrix and other markers were detected by the method of accumulated optical density(IOD) of immunohistochemistry.Using the software of Image J for statistics,the effects of cantharidin,cimicin and the combination of drugs on the above links were observed.Bleomycin can promote the production of col1,the main component of ECM,and α-SMA,the marker of myofibroblasts,to form fibrosis.(1)Compared with BLM group,the expression of col1 and α-SMA in lung tissue was decreased in awn handle group,cimicin group,combination group and PFD group.The difference was statistically significant.(P < 0.01)(2)the inhibitory effect of the combined group on the expression of col1 and α-SMA was significantly better than that of the cimicin group(P < 0.05),but there was no significant difference between the combined group and the awn handle group.(3)The inhibition of collagen deposition and extracellular matrix formation in PFD group was significantly better than that in cimicin group(P < 0.05),but the inhibition of col1 expression in PFD group was weaker than that in awn handle group(P < 0.05)Conclusion: 1.Emodin,the effective component of Astragalus membranaceus,can inhibit the inflammation of lung tissue,slow down the damage of lung tissue,improve the lung function of mice,alleviate the bleomycin induced pulmonary fibrosis of mice,and is expected to become a new drug for the treatment of pulmonary fibrosis.2.2.There was no significant difference in the pharmacodynamic effect of the combination of cantharidin and cimicin,but it was better than that of cimicin alone,suggesting that cantharidin was the main anti fibrosis drug in the combination.3.Emodin and cimicin can inhibit the proliferation of fibroblasts and the formation of extracellular matrix by reducing the deposition of collagen in lung tissue 4.The efficacy of the combination of monomers can not be overlapped and can not replace the compatibility of traditional Chinese medicine.