| Objective Based on the key factor of imbalance of T helper cell 1(Th1)and T helper cell 2(Th2)causing radiation pulmonary fibrosis(RPF),this study aims to explore the effective mechanism of Sang Xing Lung-protection particles(SX)in preventing and treating radiation pulmonary fibrosis(RPF).Method This experiment randomly divided 384 male SD rats into 6 groups,including blank group(given normal saline 10ml/d),model group(given normal saline 10ml/d),hormone group(given prednisone tablets aqueous solution with the concentration of 0.2mg/ml,15.6ml/Kg,10ml/d),Sang Xing lung-protection Particles groups(given SX concentrate with the concentration of 2g/ml,5.6ml/Kg,11.25ml/Kg,22.5ml/Kg,10ml/d),excep for the blank group,the other groups were given X ray chest single irradiation 14 Gy,all the animals were given with gavage once a day.At designated time(4th,8th,12 nd,16th,20 th,24th,28 th,32nd week)8 rats from each group were randomly executed,acquired lung tissue and 5 ml blood from the heart.The pathological changes of lung tissue was observed by HE and Masson staining,the content of plasma IL-6,TNF-α,SOD,MDA,IFN-γ,IL-4,IL-5,IL-12 and IL-13 changes were detected by ELISA assay,immunohistochemical staining was used to detect the changes of lung tissue TGF-β1 and α-SMA content.The Real-time PCR and assay was used to detected the m RNA of IL-12,IFN-γ,IL-4,IL-5 and IL-13 in lung tissue,and the flow cytometry(FCM)assay was used to detected the ratio of Th1 and Th2 cells.Results 1.The development of the RILI: at 16 th week after irradiation,the model group rats began to present the pulmonary fibrosis;at 24 th week after irradiation,the model group rats mainly presented the pulmonary fibrosis pathology;at 32 nd week after irradiation,the pulmonary fibrosis pathology of the model group rats exacerbated.It was suggested that 14 Gy irradiation agent could successfully establish the rats model of radiation induced lung injury(RILI)and radiation pulmonary fibrosis(RPF),and the model was similar to the clinic.2.Pathological: compared with the blank group,other groups rats with HE and Masson staining were observed destroyed lung tissue alveolar structure,inflammatory cell infiltration and collagen fibers with varying degrees.With the development of the time,compared with the model group,there was obvious amelioration in the inflammatory cell infiltration and pulmonary fibrosis with varying degrees in other medicine groups rats(p<0.05);compared with the hormone group,there was obvious difference before 16 th week and no difference at 20 th,24th,28 th and 32 nd weeks;there was no difference between the SX groups.3.Compared with the blank group,the content of IL-6,TNF-α,MDA in serum and TGF-β1,α-SMA in lung tissue of the model group increased(p<0.05),the activity of SOD decreased(p<0.05);compared with the model group,the content of IL-6,TNF-α,MDA,TGF-β1 and α-SMA of medicine groups decreased(p<0.05),the activity of SOD increased(p<0.05);compared with the hormone group,before 16 th week,the content of IL-6,TNF-α,MDA,TGF-β1 and α-SMA of the SX groups increased(p<0.05),the activity of SOD decreased(p<0.05),and there was no difference at 20 th,24th,28 th and 32 nd weeks;there was no difference between the SX groups.4.Compared with the blank group,the content of IL-12 and IFN-γ in serum of the model group decreased(p<0.05),the content of IL-4,IL-5 and IL-13 increased(p<0.05);compared with the model group,the content of IL-12 and IFN-γ of medicine groups increased(p<0.05),the content of IL-4,IL-5 and IL-13 decreased(p<0.05);compared with the hormone group,at 12 nd,16th weeks,the content of IL-12 and IFN-γ of the SX groups decreased(p<0.05),the content of IL-4,IL-5 and IL-13 increased(p<0.05),and there was no difference at 20 th,24th,28 th and 32 nd weeks;there was no difference between the SX groups.5.Compared with the blank group,the content of IL-12 m RNA and IFN-γm RNA expression in lung tissue of the model group decreased(p<0.05),the the content of IL-4m RNA,IL-5m RNA and IL-13 m RNA expression increased(p <0.05);compared with the model group,the content of IL-12 m RNA and IFN-γm RNA expression of medicine groups increased(p<0.05),the IL-4 m RNA,IL-5 m RNA and IL-13 m RNA decreased expression(p<0.05);compared with the hormone group,at 4th,16 th weeks,the content of IL-12 m RNA and IFN-γm RNA expression of the SX groups decreased(p<0.05),the content of IL-4 m RNA,IL-5 m RNA and IL-13 m RNA expression increased(p<0.05),and there was no difference at 20 th,24th and 32 nd weeks;there was no difference between the SX groups.6.Compared with the blank group,the ratio of the Th1 and Th2 cells of the model group decreased(p<0.05);compared with the model group,the ratio of the Th1 and Th2 cells of medicine groups increased(p<0.05);compared with the hormone group,at 12 nd,16th weeks,the ratio decreased(p<0.05),and there was no difference at 20 th,24th,28 th and 32 nd weeks;there was no difference between the SX groups.Conclusion 1.Sang Xing Lung-Protection Particles(SX)could reduce inflammatory reaction and degree of Pulmonary fibrosis of the radiation induced lung injury(RPF)rats.2.SX could reduce the content of IL-6,TNF-α,MDA,TGF-β1,α-SMA and increase the activity of SOD,this might be one of the effective mechanism of SX in preventing and treating RPF.3.The decreased expression of Th1 and increased expression of Th2 were closely related to RPF,the unbalance of Th1 and Th2 expression might be the key mechanism of RPF.4.SX could increase Th1 expression and decreased Th2 overexpression,it was proved that SX could regulate the balance of Th1 and Th2,this might be the key mechanism of SX preventing and treating RPF. |
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