| Objective This experimental research make TGF-beta 1 gene specificity silence with RNA interference,proving compound MaXingShiGan soup regulate and control radioactive lung injury TGF-beta/smad signaling pathways.Methods 1.Total of 18 clean sd rats were randomly divided into two groups,Chinese medicine group and normal control group,with 12 rats and 6 rats.According to the clinical adult dose of 15 times the preparation of traditional Chinese medicine compound containing medicine serum.giving sd rats gavage twice a day for three days,collect blood after last 1-2h.2.The total dose of radiation to cell is 8Gy,frequency 3.64Gy/min.3.After subculture,using RNA interference specificity silence TGF-beta 1 gene.4.The expressions of cell viability were analyzed by cell counting kit-8,cell apoptosis were analyzed by flow cytometry instrument,the level of TGF-beta land Smad were analyzed by PCR and Western-blot.Results 1.Compared with blank control group and Negative control group,the interference of TGF-beta 1 mRNA were reduced(P<0.05),and TGF-beta 1 1608 was the most efficient.2.After 24h、48h、72h,compared with blank control group,the group of siRNA cell viability were reduced(P<0.05),compared with siRNA group,the group of traditional Chinese medicine cell viability were reduced(P<0.05).3.The group of siRNA cell apoptosis were reduced without statistical significance compared with blank control group(P>0.05),compared with siRNA group,the group of traditional Chinese medicine cell apoptosis were increased(P<0.05).4.Compared with rat serum group,the expressions of Smad2、Smad6 mRNA about siRNA group were reduced without statistical significance(P>0.05),but Smad3、TGF-beta 1 mRNA were reduced,Smad7mRNA increased with statistical significance(P<0.05).Compared with siRNA group,the expressions of Smad2、Smad3、Smad7 mRNA about traditional Chinese medicine group were increased(P<0.05)5.Compared with rat serum group,the expressions of P-Smad2、P-Smad3、Smad6、Smad7 about siRNA group were reduced with statistical significance(P<0.05).Compared with siRNA group,the traditional Chinese medicine group expressions of P-Smad2、P-Smad3 reduced while Smad6、Smad7 increased(P<0.05).Conclusion 1.The RNA interference targeting TGF-beta1 gene can effectively suppress the expression of TGF-betalmRNA.2.The alveolar type Ⅱ cells after radiotherapy were restrained with Gene Silencing TGF-beta1,but traditional Chinese medicine may still restrain proliferation through other pathway and target spot,with due to the transfection is not completely.3.Silence TGF-beta 1 gene had no effect on alveolar type Ⅱ cell apoptosis after radiotherapy,but traditional Chinese medicine may still Promote apoptosis through other pathway or target spot,with due to the transfection is not completely.4.Silence TGF-beta 1 block radioactive lung injury TGF-beta/smad signaling pathways,but traditional Chinese medicine may still through other pathway signal transduction,with due to the transfection is not completely. |
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