加味麻杏石甘汤对放射性肺损伤TGF-β/Smad信号通路调控的研究  

Validation of the Effect of Modified Maxingshigan Decoction on the TGF-β/Smad Signaling Pathway in Radioactive Lung Injury

作　　者：林胜友[1] 周罗瑜[2] 许远[2] 

机构地区：[1]杭州市肿瘤医院,杭州310002 [2]浙江中医药大学

出　　处：《浙江中医药大学学报》2015年第12期843-848,853,共7页Journal of Zhejiang Chinese Medical University

基　　金：国家自然科学基金资助项目(81173246)~~

摘　　要：[目的]研究加味麻杏石甘汤对TGF-β/Smad信号通路的调控,阐述其抗放射性肺损伤的机制。[方法](1)清洁级SD大鼠分为中药组和正常对照组,中药组加味麻杏石甘汤灌胃3d后制备含药血清,正常对照组生理盐水灌胃制备大鼠血清;(2)小鼠肺泡Ⅱ型细胞培养,射线照射(频率为3.64Gy/min,总剂量为8Gy)造模,含药血清干预;(3)RNA干扰特异性沉默TGF-β1基因后,PCR检测TGF-β1、PAI-1、CTGFm RNA,Western-blot检测Smad蛋白水平,并行统计学比较。[结果](1)与大鼠血清组比较,10%含药血清组TGF-β1m RNA、P-Smad2表达下调(P<0.05);(2)沉默TGF-β1基因后,TGF-β1m RNA表达及其下游靶基因PAI-1m RNA、CTGF m RNA均出现一致性低表达(P<0.05),均明显低于未沉默TGF-β1基因的大鼠血清组(P<0.05);(3)10%含药血清+si RNA组与大鼠血清+si RNA组比较,不论是P-Smad2、P-Smad3,还是下游靶基因PAI-1m RNA、CTGF m RNA表达差异未见统计学意义(P>0.05);(4)10%含药血清+si RNA组Smad6、Smad7蛋白表达较大鼠血清+si RNA组均上升,差异有统计学意义(P<0.05,P<0.01)。[结论]加味麻杏石甘汤抵抗放射性肺损伤机制可能与干预TGF-β/Smad信号通路有关。[Objective] To prove the modified Maxingshigan Decoction（modified MXSGD） intervening radiation induced lung injury（RILI） through the TGF-β/Smad signaling pathway by selectively silencing the TGF-β1 gene. [Methods]（1）Total of 18 clean SD rats were randomly divided into two groups：Chinese medicine group treated with modified MXSGD and normal control group treated with saline. The serum was made after 3 days of taking drugs to prepare medicated serum.（2） The alveolar type Ⅱcells were cultured. The total dose of radiation to cell was 8Gy, frequency 3.64Gy/min. The medicated serum was given to each group respectively.（3） After silencing TGF-β1gene with RNA interference technology, the level of TGF-β1, PAI-1, CTGFm RNA was analyzed by PCR and the protein expression of Smads was measured by Western-blot. [Results]（1） Compared with rat serum group, the expressions of TGF-β1m RNA, P-Smad2 in 10% medicated serum group were down-regulation or deceased（P〈0.05）.（2） Compared with rat serum group, the transcription levels of TGF-β1, and PAI-1 and CTGF were obviously suppressed after silencing TGF-β gene（P〈0.05）.（3） There was no significance effect on the expression of P-Smad2, P-Smad3, PAI-1m RNA and CTGF m RNA in both 10% medicated serum ＋si RNA group and rat serum＋si RNA group（P〈0.05）.（4）Compared with rat serum＋si RNA, the expression of Smad6 and Smad7 was increased in 10% medicated serum＋si RNA group. The differences between the two groups were significant（P〈0.05, P〈0.01）. [Conclusion] Modified MXSGD may intervene the RILI by regulating TGF-β/Smad signaling pathway.

关 键 词：放射性肺损伤 加味麻杏石甘汤 RNA干扰 TGF-Β/SMAD信号通路 

分 类 号：R273[医药卫生—中西医结合]