| Objective:The model of radiation-induced lung injury(RILI)was established by selecting appropriate radiation dose,The effects of TanshinoneⅡA combined with umbilical cord mesenchymal stem cells on body weight,appearance,expression of Smad3p-smad3 Smad7 and TGF-β1 protein,serum cytokines and lung histopathology of RILI rats were observed.Methods:(1)In the first part of the experiment,the rats in the same group were irradiated with different doses of 60 Coγ rays of 15,18 and 20 Gy respectively,and the rat RILI model was established by selecting appropriate doses.(2)In the second part of the experiment,60 adult female SD rats of SPF grade were treated with whole chest 18 Gy single irradiation except 12 sham operated rats;After modeling,the rats were given sham operation group(12 rats in group A,intraperitoneal and caudal vein injection of normal saline),model control group(12 rats in group B,intraperitoneal and caudal vein injection of normal saline),Tanshinone IIA group(12 rats in group C,Tanshinone IIA injection in abdominal cavity,physiological saline injection in tail vein),HUMSCs group(12 rats in Group D,intraperitoneal injection of normal saline,tail vein injection of HUMSCs),Tanshinone IIA combined with HUMSCs group(12 rats in Group E,Tanshinone IIA injection and HUMSCs injection in caudal vein),and each group was injected with 15 ml /kg /d for 30 days after modeling,and 1ml/d for a single injection after modeling.Six rats in each group were randomly selected on the 30 th and 60 th day to be killed and their body weight and appearance scores were measured;the expression of TGF-β1 and IL-6 in serum of rats were detected by enzyme-linked immunosorbent assay(ELISA);the lung tissues were stained by He and Masson;Western blot The expression levels of Smad3,p-smad3,Smad7 and TGF-β1 protein in the lung tissue of rats were detected by WB.The effect of TanshinoneⅡA combined with HUMSCs on RILI in rats was studied.Results:(1)The appropriate dose of 18 Gy was selected to establish the rat model of radiation-induced lung injury.(2)Comparison of weight evaluation of rats in each group: before the experiment,there was no statistical difference in the weight of rats in each group(P > 0.05).Compared with sham operation group,the body weight of model group,Tanshinone IIA group,HUMSCs group,Tanshinone IIA combined with HUMSCs group was significantly lower than that of sham operation group(P < 0.05).Compared with the model group,on the 30 th and 60 th day,the body weight of sham operation group,Tanshinone IIA group,HUMSCs group,Tanshinone IIA combined with HUMSCs group was significantly higher than that of the model group(P < 0.05).There was no significant difference in body weight between Tanshinone IIA group,HUMSCs group,Tanshinone IIA and HUMSCs group on the 30 th day(P > 0.05).On the 60 th day,the body weight of Tanshinone IIA combined with HUMSCs group was significantly higher than that of Tanshinone IIA group and HUMSCs group(P < 0.05),and there was no significant difference between Tanshinone IIA group and HUMSCs group on the 60 th day(P > 0.05).(3)Comparison of appearance scores of rats in each group: at the 30 th day,the scores of rats in the sham operation group were significantly higher than those in the model group,Tanshinone IIA group,HUMSCs group and Tanshinone IIA combined with HUMSCs group(P < 0.05).The score of model group was significantly lower than that of sham operation group,Tanshinone IIA group,HUMSCs group and Tanshinone IIA combined with HUMSCs group(P < 0.05).There was no significant difference in scores among Tanshinone IIA group,HUMSCs group and Tanshinone IIA combined with HUMSCs group(P > 0.05).At the 60 th day,the scores of each group were significantly different,from high to low: sham operation group,Tanshinone IIA combined with HUMSCs group,Tanshinone IIA group,HUMSCs group,model group,the differences were statistically significant(P < 0.05).(4)Comparison of pathological evaluation of lung tissue in each group: at the 30 th day,the inflammatory score of lung tissue in each group was from high to low,which were model group,Tanshinone IIA group,HUMSCs group,Tanshinone IIA combined with HUMSCs group.There was no statistical difference between Tanshinone IIA group and HUMSCs group(P > 0.05),and the difference between the other groups was statistically significant(P <0.05).On the 60 th day,the inflammatory scores of model group,Tanshinone IIA group and HUMSCs group were significantly higher than thoseof Tanshinone IIA combined with HUMSCs group(P < 0.05).There was no significant difference among model group,Tanshinone IIA group and HUMSCs group(P > 0.05).In terms of pulmonary fibrosis,at the 30 th and 60 th day,the fibrosis scores were from high to low in the model group,Tanshinone IIA group,HUMSCs group,Tanshinone IIA combined with HUMSCs group,and the difference was statistically significant(P <0.05).(5)Comparison of serum and IL-6 levels of rats in each group: on the 30 th day of this experiment,the levels of TGF-β1 and IL-6 in each group were from high to low,which were respectively model group,Tanshinone IIA group,HUMSCs group,Tanshinone IIA combined with HUMSCs group and sham operation group.On the60 th day of the experiment,the levels of TGF-β1 in each group were from high to low,which were model group,Tanshinone IIA group,HUMSCs group,Tanshinone IIA combined with HUMSCs group,sham operation group There was no significant difference in the level of IL-6 between the two groups.(6)Comparison of the expression levels of Smad3,p-smad3,Smad7 and TGF-β1protein in the lung tissues of each group: on the 30 th and 60 th day of the experiment,the expression levels of TGF-β1,Smad3 and P-smad3 protein in each group were from high to low,which were model group,Tanshinone IIA group,HUMSCs group,Tanshinone IIA combined with HUMSCs group and sham operation group.The expression level of Smad7 in sham operation group,Tanshinone IIA combined with HUMSCs group,HUMSCs group,Tanshinone IIA group and model group respectively.Conclusion:(1)The radiation-induced lung injury model of rats can be established by 18 Gy radiation.(2)Tanshinone IIA,HUMSCs and the combination of the two can reduce the alveolar inflammatory response,delay the process of pulmonary fibrosis,and play a protective role in lung tissue.(3)The protective mechanism of Tanshinone IIA and HUMSCs on lung tissue after radiation exposure may be realized through several ways:(1)Down regulating TGF-β1protein expression,down regulating Smad3 and p-smad3 protein expression,upregulating Smad7 protein expression.(2)It can inhibit the release of IL-6 and TGF-β1 in serum.(4)The combination of them can further down regulate the expression of TGF-β1,Smad3 and p-smad3,and up regulate the expression of Smad7. |
PDF Full Text Request