摘要
目的探讨丹皮酚联合三七总皂苷应用对血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)诱导新生大鼠的心脏成纤维细胞(cardiac fibroblasts,CFs)增殖及TGF-β/Smads信号通路的影响。方法采用胰酶消化法分离培养的CFs随机分为6组:正常对照组,模型组,三七总皂苷组,丹皮酚组,丹皮酚与三七总皂苷联合用药大、小剂量组。采用MTT比色法测定各组培养液中CFs增殖情况,免疫细胞化学法检测α-平滑肌肌动蛋白(α-SMA)的蛋白表达水平,RTPCR法检测转化生长因子-β1(transforming growth factor-β1,Tgfb1)的mRNA表达水平,Western blot法检测TGF-β1、磷酸化的Smad2/3(p-Smad2/3)的蛋白表达水平。结果与正常对照组比较,模型组大鼠CFs增殖明显增高,Tgfb1的mRNA表达显著增加,α-SMA及TGF-β1、p-Smad2/3的蛋白表达显著增加;与模型组比较,丹皮酚组、三七总皂苷组、联合用药的大剂量和小剂量组均可抑制由AngⅡ诱导的CFs的增殖,Tgfb1的mRNA表达显著减少,α-SMA及TGF-β1、p-Smad2/3的蛋白表达显著减少;与丹皮酚组、三七总皂苷组比较,联合用药大、小剂量组在抑制CFs增殖的作用上显著增强,抑制Tgfb1的mRNA表达效果显著增强,抑制α-SMA及TGF-β1、p-Smad2/3的蛋白表达效果显著增强;与联合用药小剂量组比较,联合用药大剂量组对以上指标的抑制效果更佳,显示一定的剂量依赖性。结论丹皮酚与三七总皂苷联合应用对抑制CFs增殖,呈现明显的协同作用。其作用机制与抑制TGF-β/Smads信号通路的传导有关。
AIM To investigate the effects of combining paeonol( Pae) with Panax notoginseng saponins( PNS) on the proliferation of cardiac fibroblasts( CFs) induced by angiotensinⅡ( AngⅡ) and TGF-β/Smads signal pathway. METHODS The CFs of neonate rat were isolated and cultured by trypsin digestion and randomly divided into six groups: normal control group,model group,PNS group,Pae group,the combination of Pae and PNS high-dose and low-dose groups. The cell proliferations were measured with MTT. The expression of α-smooth muscle actin( α-SMA) protein was detected by immunocytochemistry method. The mRNA expression of transforming growth factor-β1( Tgfb1) was detected by RT-PCR. The protein levels of TGF-β1 and p-Smad2/3 were determined by Western blot. RESULTS Compared with the control group,the rat CFs proliferation,Tgfb1 mRNA,α-SMA protein,TGF-β1 and p-Smad2/3 protein expression increased obviously in the model group. Compared with the model group,the CFs proliferation,Tgfb1 mRNA,α-SMA protein,TGF-β1 and p-Smad2/3 protein expression decreased significantly in PNS group,Pae group,the combination of Pae and PNS high-dose and small-dose groups. Compared with PNS group and Pae group,the CFs proliferation,Tgfb1 mRNA,α-SMA protein,TGF-β1 and p-Smad2/3 protein expression decreased significantly in combination of Pae and PNS high-dose and low-dose groups. With regard to the inhibition of abovementioned indicators,the effect of Pae combined with PNS high-dose group were better than that of Pae combined with PNS low-dose group in a dose-dependent manner.CONCLUSION The combination of Pae and PNS can inhibit CFs proliferation and show significant synergistic effects related with the suppression of TGF-β/Smads signal transduction pathway.
出处
《中成药》
CAS
CSCD
北大核心
2016年第4期729-735,共7页
Chinese Traditional Patent Medicine
基金
河北省自然科学基金项目(C2011406009)
河北省教育厅重点项目(ZD20131068)