柴胡桂枝干姜汤抗胰腺纤维化作用与抑制ERK1/2通路有关

Anti-fibrosis effect of Chaihu Guizhi Ganjiang decoction related to inhibition of ERK1/2 pathway

目的:从ERK1/2信号通路角度探讨柴胡桂枝干姜汤抗胰腺纤维化的作用机制。方法:将30只健康雄性Wistar大鼠随机分为对照组、模型组、治疗组。采用一次性尾静脉注射二氯二丁基酯(DBTC)7 mg/kg制备大鼠胰腺纤维化模型。自制模后第2天,治疗组给予浓度为1.0 g/mL的柴胡桂枝干姜汤灌胃(10 g/kg体重),1次/d,直到实验结束;模型组给予等体积饮用水灌胃。各组大鼠分别于制模后28 d取胰腺组织行苦味酸-天狼星红染色观察胰腺纤维化程度,采用酶联免疫分析法(ELISA)测定胰腺组织I型和Ⅲ型胶原含量,并分别用荧光定量PCR和Western blotting检测细胞外信号调节激酶1/2(ERK1/2)和双特异性磷酸酶(DUSP)6的mRNA和蛋白表达水平。结果:模型组大鼠胰腺组织可见弥漫性纤维组织增生,Ⅰ、Ⅲ型胶原纤维交织成网络,胰腺小叶结构破坏或消失,腺泡萎缩变形;治疗组部分胰腺小叶纤维组织增生、腺泡结构破坏,尚可见正常胰腺结构。与对照组比较,模型组大鼠胰腺组织I、Ⅲ型胶原含量升高,DUSP6表达降低,p-ERK1/2表达升高,差异均有统计学意义(P<0.05);与模型组比较,治疗组大鼠胰腺组织I、Ⅲ型胶原含量减少,DUSP6表达升高,p-ERK1/2表达降低,差异均有统计学意义(P<0.05)。结论:柴胡桂枝干姜汤减少胶原生成、抗胰腺纤维化的作用机制与上调DUSP6表达、抑制ERK1/2表达和活性有关。

Objective To explore the anti-fibrosis mechanism of Chaihu Guizhi Ganjiang Decoction in the treatment of pancreatic fibrosis based on the ERK1/2 signal pathway.Methods 30 healthy male Wistar rats were randomly divided into control group,model group and treatment group.A rat model of pancreatic fibrosis was established by a single injection of DBTC(7 mg/kg)through the caudal vein.On the second day after the injection,the treatment group was given Chaihu Guizhi Ganjiang Decoction(1.0g/mL,10 g/kg body weight)once a day until the end of the experiment.The model group was given equal volume drinking water.The pathological changes of pancreas were observed by picric-sirius red staining on the 28th day after the injection.The contents of collagen type Ⅰ and type Ⅲ were determined by enzyme-linked immunosorbent assay(ELISA),and the expression levels of mRNA and protein of extracellular signal regulated kinase 1/2(ERK1/2)and dual specificity phosphatase 6(DUSP 6)were detected by fluorescence quantitative PCR and Western blotting respectively.Results Diffuse fibrous tissue hyperplasia was observed in the pancreas of the model group:collagen type Ⅰ and type Ⅲ were interweaved into a network,the structure of pancreatic lobules was destroyed or disappeared,and acini were atrophic or deformed.In the treatment group,some pancreatic lobules were proliferated and the acinar structure was destroyed.In the treatment group,the fibrous tissue hyperplasia was found in some pancreatic lobules,and the acini structure were destroyed,but the normal acini was also found in another lobules.Compared with the control group,the contents of type Ⅰ and type Ⅲ collagen were increased,the expression of DUSP6 was decreased and the expression of ERK1/2 was increased in the model group(P<0.05).Compared with the model group,the contents of type Ⅰ and type Ⅲ collagen were decreased,the expression of DUSP 6 was increased,and the expression of ERK1/2 was decreased in the treatment group(P<0.05).Conclusion The anti-fibrosis mechanism of Chaihu Guizhi Ganjiang Decoction is related to the up-regulation of DUSP 6 expression and inhibition of ERK1/2 expression and activity in chronic pancreatitis.