大黄丹参水煎剂对胰腺纤维化大鼠转化生长因子β_1、血小板衍生生长因子BB及其mRNA表达的影响

Effects of Dahuang Danshen Decoction on the Expressions of TGF-β_1,PDGF-BB,TGF-β_1 mRNA and PDGF-BB mRNA in Rats With Pancreatic Fibrosis

目的观察大黄丹参水煎剂抗胰腺纤维化的可能作用机制。方法 50只雄性Wistar大鼠随机分为正常组、模型组和大黄丹参水煎剂低、中、高剂量组,每组10只。腹腔注射二乙基二硫代氨基甲酸钠(DDC)6周建立胰腺纤维化模型。大黄丹参水煎剂低、中、高剂量组在造模同时分别给予大黄丹参水煎剂0.137、0.274、0.548 g/(kg·d)灌胃,正常组和模型组给予等量生理盐水灌胃,连续给药6周。免疫组化检测胰腺α-平滑肌肌动蛋白(α-SMA)表达,RT-PCR检测胰腺大鼠转化生长因子β1(TGF-β1)mRNA、血小板衍生生长因子BB(PDGF-BB)mRNA表达,免疫印迹法检测TGF-β1蛋白、PDGF-BB蛋白表达。结果正常组α-SMA低表达,模型组α-SMA高表达,大黄丹参水煎液各剂量组α-SMA均有不同程度的减弱。模型组TGF-β1、PDGF-BB蛋白B mRNA表达较正常组上调(P<0.05或P<0.01);与模型组比较,大黄丹参中、高剂量组TGF-β1蛋白及mRNA表达下调,大黄丹参高剂量组PDGF-BB蛋白及mRNA下调(P<0.05或P<0.01)。与大黄丹参中剂量组比较,大黄丹参高剂量组TGF-β1蛋白下降显著(P<0.05)。结论"下调TGF-β1及PDGF-BB及其mRNA表达—抑制α-SMA表达—抑制PSC活化—减少ECM合成—改善ECM沉积"可能是大黄丹参抗胰腺纤维化的作用机制之一。

Objective To observe the effect and mechanism of Decoction of Dahuang （Rhei Radix et Rhizoma） - Danshen （Salviae Miltiorrhizae Radix） on rats with pancreatic fibrosis. Methods Fifty male Wistar rats were ran- domized into the normal group, model group and high-dose, middle-dose and low-dose Dahuang-Danshen groups, with 10 in each. The pancreatic fibrosis models were established by intraperitoneal injection of diethyl dithiocarbamate （DDC） for 6 weeks. The low-dose, middle-dose and high-dose Dahuang-Danshen groups were garaged with 0. 137 g/ （kg d）, 0. 274 g/ （kg d） and 0. 548 g/ （kg d） Dahuang-Danshen respectively. The normal group and model group were garaged with same volume of normal saline. The administration lasted for 6 weeks. The expressions of α- SMA, transforming growth factor-β1 （TGF-β1） TGF-β1 mRNA, platelet-derived growth factor-BB （PDGF-BB） and PDGF-BB mRNA in pancreas were detected with immunohistochemical detection, RT-PCR and Western Blotting re- spectively. Results The expression of α-SMA was low in the normal group but high in the model group. The expression of α-SMA was decreased in the Dahuang-Danshen groups. Comparing with the normal group, the expressions of TGF-β1 mRNA and PDGF-BB mRNA were significantly increased in the model group （ P 〈 0.05 ）. Comparing with the model group, the expression of TGF-β1 mRNA was significantly decreased in middle-dose and high-dose Dahuang-Danshen groups and the expression of PDGF-BB mRNA was significantly decreased in high-dose Dahuang- Danshen group （P 〈0.05）. Comparing with middle-dose Dahuang-Danshen group, the expression of TGF-β1 was significantly decreased in high-dose Dahuang-Danshen group （P 〈 0.05）. Conclusion The anti-pancreatic fibrosis mainline of Dahuang-Danshen may be down-regulating the expressions of TGF-β1, TGF-β1 mRNA, PDGF and PDGF mRNA, inhibiting the expression of α-SMA, inhibiting the activation of PSC, reducing the synthesis of ECM and im- proving the deposition of ECM.