摘要
目的 探讨乙氧基血根碱对血管紧张素Ⅱ(AngⅡ)诱导的小鼠心肌纤维化的影响及作用机制.方法 构建AngⅡ诱导的高血压小鼠心肌纤维化模型,根据小鼠的基线血压随机分为空白组、模型组、低剂量组、中剂量组、高剂量组、阳性药组.除空白组之外的其余各组小鼠皮下以500 ng/(kg·min)的速率释放AngⅡ,空白组在泵内注入等量体积生理盐水,持续4周;低、中、高剂量组从术后第1天起分别给予0.1、1、10 mg/(kg·d)乙氧基血根碱灌胃,空白组和模型组给予等体积生理盐水,阳性药组给予10 mg/(kg·d)缬沙坦,连续灌胃给药28 d.采用Masson染色及天狼星红染色法检测6组小鼠心肌纤维化及胶原沉积情况;Western blot检测Ⅰ型胶原、Ⅲ型胶原、增殖细胞核抗原(PCNA)、α-平滑肌肌动蛋白(α-SMA)、纤维连接蛋白(FN)及TGF-β1/smad2/3信号通路相关蛋白TGF-β1、p-smad2/3、smad2/3的蛋白表达量.结果 与空白组比较,模型组小鼠心肌细胞出现明显纤维化及胶原沉积,心脏组织中Ⅰ型胶原、Ⅲ型胶原、PCNA、α-SMA及FN的蛋白表达显著增加(P＜0.05),通路相关蛋白TGF-β1表达、p-smad2/smad2比值及p-smad3/smad3比值显著上调(P＜0.05),而乙氧基血根碱给药后,心肌的纤维化及胶原沉积情况均明显改善,Ⅰ型胶原、Ⅲ型胶原、PCNA、α-SMA、FN、TGF-β1等蛋白表达及p-smad2/smad2和p-smad3/smad3比值均显著下调(P＜0.05),且乙氧基血根碱对小鼠心肌纤维化改善的作用呈剂量依赖性(P＜0.05).结论 乙氧基血根碱可改善AngⅡ诱导的心肌纤维化,通过抑制TGF-β/smad2/3通路的活化可能是其发挥作用的机制之一.
Abstract
Objective:To investigate the effect and mechanism of Ethoxysanguinarine(ETH)on cardiac fibrosis induced by An-giotensin Ⅱ(Ang Ⅱ)in mice.Methods:Cardiac fibrosis model of hypertensive mice induced by AngⅡwas established.The mice were randomly divided into blank group,model group,ETH-low,ETH-medium and ETH-high groups,positive drug group.ALL the mice except the blank group were injected with Ang Ⅱ at a rate of 500 ng/(kg·min)subcutaneously while the blank group was injected with the same volume of normal saline for four weeks.From the first day after surgery,the ETH-low,ETH-medium and ETH-high groups were given 0.1,1,and 10 mg/(kg·d)of ETH by gavage.Respectively,the blank group and the model group were given the same volume of normal saline,and the positive drug group was given 10 mg/(kg·d)of Valsartan.All groups were given drugs by con-tinuous gavage for 28 days.Masson staining and Sirius red staining were used to detect cardiac fibrosis and collagen deposition in mice.The expression of collagen I,collagen Ⅲ,proliferating cell nuclear antigen(PCNA),α-smooth muscle actin(α-SMA),fibro-nectin(FN)and TGF-β1/smad2/3 signaling pathway-related proteins TGF-β1,p-smad2,smad2,p-smad3 and smad3 were detected by Western blot.Results:Compared with the blank group,there was significant cardiac fibrosis and collagen deposition in mice car-diomyocytes in the model group,the protein expression of collagen-I,collagen Ⅲ,PCNA,α-SMA,FN,TGF-β1,p-smad2,smad2,p-smad3 and smad3 ratio in heart tissue were significantly up-regulated(P<0.05),which were significantly reverssed after ETH accom-panied with alleviation of cardiac fibrosis and collagen deposition in a dose-dependent manner(P<0.05).Conclusion:ETH can alleviate AngⅡ-induced cardiac fibrosis,the inhibition of TGF-β1/smad2/3 pathway may be one of the important mechanisms.
基金项目
福建省自然科学基金(2021J01940)
国家自然科学基金(U22A20372)
中华中医药学会青年人才托举工程项目(2021-QNRC2-B19)
福建中医药大学青年科研拔尖人才项目(XQB202202)
维普
万方数据