| Objective:To establish model rats with pulmonary fibrosis and treating this disease with prednisone acetate and different doses of Ophiopogon Decoction to observe the mechanism of Ophiopogon Decoction against pulmonary fibrosis.Methods:?Grouping:120 healthy SD rats were randomly divided into normal group,model group,prednisone acetate group,low dose of Ophiopogon Decoction group,middle dose group of Ophiopogon Decoction group,and high dose group of Ophiopogon Decoction group,with 20 rats in each group.?Modeling:Mo del rats with pulmonary fibrosis were established with tracheal infusion.Model group and all medical groups were treated with tracheal infusion of bleomycin 0.lml/100g(5mg/kg),and the normal group was perfused with tracheal saline 0.1 ml/100g.?Medication:After modeling,all medical groups were treating with Ophiopogon Decoction by gavage in the second day.The normal group and the model group were treated with deionized water.The prednisone acetate group was treated with prednisone acetate saline solution and the dosage was 3.15 mg/kg.The rats in the low dose group were treated with low dose of 7.29 g crude drug/kg and the dose of 21 mg/For 36.45g crude drug/kg.Each group of rats were given 2ml/100g,once a day.After 14 days of continuous administration,10 animals in each group were randomly sacrificed,and the remaining animals continued with gavage until the 28th day.?After the last administration,the rats were fasting for 24 hours without water,and they were narcotized with 4%chloral hydrate solution(3ml/kg)by intraperitoneal injection anesthesia.Taking blood by puncture abdominal aorta and taking complete lung tissue in detection.The left lung was fixed with 4%POM solution,the right lung was frozen with liquid nitrogen.The levels of IFN-? and IL-4 in lung tissue of experimental animals were determined by enzyme-linked immunosorbent assay(ELISA).The expressions of TGF-? 1 and IL-4 in lung tissue were observed by immunohistochemistry.The expression of TGF-? 1 mRNA was detected by RT-PCR.? Statistic:SPSS21.0 statistical analysis software was used for data processing.Results:? HE staining:The lung tissue structure of normal group rats in the 14th days and 28th days was clear,no alveolar inflammation or fibrosis,no obvious oozing in alveolar cavity.In the model group,the alveolar septum was widened and the infiltration of inflammatory cells such as monocytes,lymphocytes and macrophages was observed and the proliferation of fibroblasts was observed on the 14th day.On the 28th day,the alveolar disease disappeared and the alveolar wall increased thick,alveolar structure was destroyed,alveolar cavity has narrowed,part of the alveolar collapse or fusion,interstitial large number of collagen fibers deposition,pulmonary fibrosis model has been formed.?IFN-?level detection results:Compared with the normal group,IFN-? levels in the model group,prednisone acetate group,low-,middle-and high-dose Ophiopogon Decoction groups were all lower at the 14th day.The differences were statistically significant(P<0.01).At the 28th day,IFN-? level in the model group,low-and middle-dose Ophiopogon Decoction groups was lower than that in the normal group with significant difference(P<0.01),while IFN-? level in the prednisone acetate group and high-dose Ophiopogon Decoction group was lower than the normal group,but the difference wasn't statistically significant(P>0.05).Compared with the model group,IFN-? levels in the prednisone acetate group,low-,middle-and high-dose Ophiopogon Decoction groups were significantly higher(P<0.01)at the 14th,28th days,while low-and middle-dose Ophiopogon Decoction groups had lower IFN-? levels in comparison with the prednisone acetate group.The differences were statistically different(P<0.01).At the 14th and 28th days,IFN-? levels in the low-and middle-dose Ophiopogon Decoction groups were lower than those in the prednisone acetate group.The differences were statistically significant(P<0.01),while the high-dose Ophiopogon Decoction group had lower IFN-?level than the prednisone group,but the difference was not statistically different(P>0.05).Compared with the low-dose Ophiopogon Decoction group,the high-dose group had significantly higher IFN-? levels at the 14th and 28th days(P<0.01),while the middle-dose group had higher IFN-? levels with no statistical difference(P>0.05).Compared with the middle-dose group,the high-dose one had significantly higher IFN-? level(P<0.01)at the 14th and 28th days.?IL-4 level detection results:Compared with the normal group,IFN-? levels in the model group,prednisone acetate group,low-,middle-and high-dose Ophiopogon Decoction groups were all higher at the 14th day.The differences were statistically significant(P<0.01).At the 28th day,IL-4 level in the model group,low-and middle?dose Ophiopogon Decoction groups was higher than that in the normal group with significant difference(P<0.01),while IL-4 level in the prednisone acetate group and high-dose Ophiopogon Decoction group was higher than the normal group,but the difference wasn't statistically significant(P>0.05).Compared with the model group,IL-4 levels in the prednisone acetate group,low-,middle-and high-dose Ophiopogon Decoction groups were significantly lower(P<0.01)at the 14th,28th days.At the 14th and 28th days,IL-4 levels in the low-and middle-dose Ophiopogon Decoction groups were higher than those in the prednisone acetate group.The differences were statistically significant(P<0.01),while the high-dose Ophiopogon Decoction group had lower IL-4 level than the prednisone group,but the difference was not statistically different(P>0.05).Compared with the low-dose Ophiopogon Decoction group,the middle-and high-dose groups had significantly lower IL-4 levels at the 14th and 28th days(P<0.01),while the middle-dose group had higher IFN-?levels with no statistical difference(P>0.05).Compared with the middle-dose group,the high-dose one had significantly lower IL-4 level(P<0.01)at the 14th and 28th days.?TGF-?1 level detection results:Compared with the normal group,TGF-? 1 levels in the model group,prednisone acetate group,low-,middle-and high-dose Ophiopogon Decoction groups were all higher at the 14th day.The differences were statistically significant(P<0.01 or P<0.05).At the 28th day,TGF-? 1 level in the model group,low-,middle-and high-dose Ophiopogon Decoction groups was higher than that in the normal group.The difference was not statistically significant(P>0.05),and TGF-? 1 in the prednisone acetate group was lower than that in the normal group,with no significant difference(P>0.05).Compared with the model group,TGF-?1 levels in the prednisone acetate group,and high-dose Ophiopogon Decoction group were lower with no statistical difference(P>0.05)at the 14th day.The middle-and high-dose groups had higher TGF-? 1 level than the model group,with no significant difference(P>0.05).At the 28th day,TGF-? 1 level in the prednisone acetate group was significantly lower than that in the model group(P<0.05),while in the low-,middle-and high-dose Ophiopogon Decoction groups,TGF-? 1 level was lower than that in the model group,with no statistical difference(P>0.05).?I type collagen level detection results:Compared with the normal group,I type collagen levels in the model group,low-,middle-and high-dose Ophiopogon Decoction groups were all higher at the 14th day.The differences were statistically significant(P<0.01 or P<0.05).The prednisone acetate group and middle-dose Ophiopogon Decoction group had higher level of I type collagen than the normal group,but the difference was not statistically significant(P>0.05).At the 28th day,I type collagen level in the model group,low-and middle-dose Ophiopogon Decoction groups was higher than that in the normal group.The difference was statistically significant(P<0.01),and the level of I type collagen in the prednisone acetate group and high-dose Ophiopogon Decoction group was higher than that in the normal group,with no significant difference(P>0.05).Compared with the model group,I type collagen level in the prednisone acetate group was lower,with statistical difference(P<0.05)at the 14th day.The low-,middle-and high-dose groups had higher I type collagen level than the model group,with no significant difference(P>0.05).At the 28th day,I type collagen level in the prednisone acetate group and high-dose Ophiopogon Decoction group was significantly lower than that in the model group(P<0.01),while in the low-and middle-dose Ophiopogon Decoction groups,I type collagen level was lower than that in the model group,with no statistical difference(P>0.05).Compared with the prednisone acetate group,I type collagen level in the high-dose Ophiopogon Decoction group was lower,with no statistical difference(P>0.05)at the 28th day.?TGF-?1 mRNA expression detection results:Compared with the normal group,TGF-?1 mRNA expression in the model group,middle-and high-dose Ophiopogon Decoction groups were all higher at the 14th day.The differences were statistically significant(P<0.01 or P<0.05).The prednisone acetate group and low-dose Ophiopogon Decoction group had higher level of TGF-? 1 mRNA expression than the normal group,but the difference was not statistically significant(P>0.05).At the 28th day,TGF-? 1 mRNA expression in the model group,prednisone acetate group and middle-dose Ophiopogon Decoction groups was higher than that in the normal group.The difference was statistically significant(P<0.01),and the TGF-? 1 mRNA expression in the low-and high-dose Ophiopogon Decoction group was higher than that in the normal group,with no significant difference(P>0.05).Compared with the model group,TGF-? 1 mRNA expression in the prednisone acetate group and high-dose Ophiopogon Decoction group was lower,with statistical difference(P<0.01 or P<0.05)at the 14th day.The low-and middle-dose groups had lower TGF-? 1 mRNA expression than the model group,with no significant difference(P>0.05).At the 28th day,TGF-? 1 mRNA expression in the prednisone acetate group,middle-and high-dose Ophiopogon Decoction group was significantly lower than that in the model group(P<0.01),while in the low-dose Ophiopogon Decoction group,TGF-? 1 mRNA expression was lower than that in the model group,with no statistical difference(P>0.05).Compared with the prednisone acetate group,TGF-? 1 mRNA expression in the high-dose Ophiopogon Decoction group was lower,with no statistical difference(P>0.05)at the 14th day.The middle-and high-dose Ophiopogon Decoction groups had lower TGF-? 1 mRNA expression than the prednisone acetate group,with significant difference(P<0.05)at the 28th day.Compared with the middle-dose Ophiopogon Decoction group,the high-dose group had significantly lower TGF-? 1 mRNA expression(P<0.01).Conclusion:1 The Ophiopogon Decoction have effects on the bleomycin induced pulmonary fibrosis in rats.2 The decoction can enhance the IFN-? expression and decrease the expression of IL-4,indicating that the decoction possibly intervene in the development process of bleomycin induced pulmonary fibrosis and 9 inflammation of pulmonary alveoli through rectifying the balance of Th1/Th2 type cytokines.3 The decoction can enhance the IFN-? expression and decrease the expression of IL-4,indicating that the decoction possibly intervene in the development process of bleomycin induced pulmonary fibrosis and inflammation of pulmonary alveoli through rectifying the balance of Th1/Th2 type cytokines.4 The high-dose Ophiopogon decoction has the best effect. |
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