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[目的]观测补中益气汤灌胃大鼠血清对TGF-β1介导A549细胞MRP1表达影响。[方法]血清药理学法,制备灌胃大鼠血清:使用随机平行对照方法,将20只SPF级SD大鼠,按随机数字表法分为2组,10只/组,补中益气汤组以2m L生药量5.67g/kg灌胃,空白血清组等量生理盐水,1次/d,连续3d,末次灌胃1h后,腹主动脉取血,提取上清。血清干预:取人肺腺癌A549细胞,接种于培养瓶中,37℃,5%CO2培养箱中培养;细胞密度达70%,饥饿12h,空白组(A组)加空白血清;TGF-β1组(B组)加空白血清及TGF-β1;补中益气汤+TGF-β1组(C组)加补中益气汤血清及TGF-β1,培养48h。观测MRP1蛋白表达(免疫细胞化学法和Western blot法)、MRP1基因表达(Realtime PCR法)。[结果]MRP1主要在细胞膜表达,各组细胞中均可见棕黄色阳性产物表达,TGF-β1组(B组)和补中益气汤+TGF-β1组(C组)棕黄色颗粒阳性表达量均高于空白组(P<0.05);补中益气汤+TGF-β1组(C组)低于TGF-β1组(B组)(P<0.05);Western blot结果与组化结果一致。Realtime PCR结果为TGF-β1组(B组)和补中益气汤+TGF-β1组(C组)基因表达水平均高于空白组(P<0.05);补中益气汤+TGF-β1组(C组)低于TGF-β1组(B组)(P<0.05)。[结论]补中益气汤血清干预TGF-β1诱导A549细胞EMT过程,细胞中MRP1在蛋白和基因水平均降低。
[Objective] To observe the effect of Buzhong Yiqi decoction fed rat serum on the expression of MRP1 in A549 cells induced by TGF-β1. [Method] Serum pharmacology method was used to prepare the serum of intragastric administration rats: 20 SPF SD rats were randomly divided into 2 groups according to random number table method, 10 rats in each group. Buzhong Yiqi Decoction The rats were gavaged with 2 ml of crude drug 5.67 g / kg, and the blank serum was injected with normal saline once a day for 3 days. After the last gavage for 1 hour, the abdominal aorta was taken and the supernatant was extracted. Serum intervention: Human lung adenocarcinoma A549 cells were inoculated into culture flasks and cultured in 37 ° C, 5% CO2 incubator; cell density was 70%, starvation 12h, blank group (group A) plus blank serum; TGF- Group B (B) plus serum and TGF-β1; Buzhong Yiqi Decoction + TGF-β1 group (C group) plus Bu Zhong Yi Tang serum and TGF-β1, cultured for 48 hours. MRP1 protein expression (immunocytochemistry and Western blot method), MRP1 gene expression (Realtime PCR method). [Results] MRP1 was mainly expressed in the cell membrane. The expression of brown-yellow positive product was observed in all the cells. The positive expression of brown granules in TGF-β1 group (B group) and Buzhong Yiqi decoction + TGF-β1 group (P <0.05). The Buzhong Yiqi Decoction + TGF-β1 group (C group) was lower than the TGF-β1 group (B group) (P <0.05). Western blot results were consistent with the histochemical results. Realtime PCR results showed that the gene expression levels of TGF-β1 group (B group) and Buzhong Yiqi decoction group + TGF-β1 group (C group) were higher than that of blank group (P <0.05) Group C (group C) was lower than TGF-β1 group (group B) (P <0.05). [Conclusion] The serum of Buzhong Yiqi decoction intervened the EMT process of A549 cells induced by TGF-β1, and the protein and gene level of MRP1 decreased in the cells.