摘要
目的:观察小陷胸汤对5-氟尿嘧啶(5-FU)介导小鼠心肌纤维化及缺氧诱导因子-1α(HIF-1α)/血管内皮生长因子(VEGF)信号通路的影响,探讨其减轻5-FU心脏损伤的可能机制。方法:将100只SPF级雄性ICR小鼠随机分为正常组、模型组、小陷胸汤低、中、高剂量(8.3、16.6、33.2 g·kg^(-1))组,每组20只。除正常组外,其余各组均采用腹腔注射5-FU(30 mg·kg^(-1))连续7 d建立心肌损伤模型小鼠。小陷胸汤低、中、高组造模同时灌胃相应剂量小陷胸汤,正常组和模型组给予等容积生理盐水,连续处理7 d。采用彩色多普勒超声心动仪检测左心室舒张末期内径(LVIDd)、左室收缩末期内径(LVIDs)、左室射血分数(LVEF)、左室短轴缩短率(LVFS);苏木素-伊红(HE)及马松(Masson)染色观察心肌组织病理形态学改变;免疫组化法测定心肌组织α-平滑肌肌动蛋白(α-SMA)表达;采用酶联免疫吸附测定法(ELISA)检测血清丙二醛(MDA)、超氧化物歧化酶(SOD)水平;实时荧光定量聚合酶链式反应(Real-time PCR)检测心肌组织HIF-1α、VEGF mRNA表达;蛋白免疫印迹法(Western blot)检测心肌组织HIF-1α、VEGF、Ⅰ型胶原蛋白(ColⅠ)及Ⅲ型胶原蛋白(ColⅢ)蛋白的表达。结果:与正常组比较,模型组小鼠LVIDd、LVIDs显著升高(P<0.01),LVEF、LVFS显著降低(P<0.01),心肌组织出现明显排列紊乱、炎性细胞浸润及间质纤维化等病理变化,心肌胶原容积分数%(CVF%)显著升高(P<0.01),α-SMA、ColⅠ、ColⅢ蛋白表达显著升高(P<0.01),MDA水平升高(P<0.01),SOD水平降低(P<0.01),HIF-1α、VEGF mRNA及蛋白表达显著升高(P<0.01)。与模型组比较,小陷胸汤中、高剂量组LVIDd、LVIDs明显降低(P<0.05,P<0.01),LVEF、LVFS明显升高(P<0.05,P<0.01),小陷胸汤各剂量组心肌组织损伤、炎性细胞浸润与心肌纤维化均明显减轻,CVF%明显降低(P<0.05,P<0.01),小陷胸汤中、高剂量组α-SMA、ColⅠ、ColⅢ蛋白表达显著降低(P<0.01),MDA显著
Objective:To observe the impact of Xiao Xianxiongtang on 5-fluorouracil(5-FU)-induced myocardial fibrosis and the hypoxia-inducible factor-1α(HIF-1α)/vascular endothelial growth factor(VEGF)signaling pathway in mice and explore its potential mechanism in suppressing 5-FU-induced cardiotoxicity.Method:One hundred SPF-grade male ICR mice were randomly divided into normal control group,model group,and Xiao Xianxiongtang groups with low-,medium-,and high-doses(8.3,16.6,and 33.2 g·kg^(-1)),with 20 mice in each group.Except for the normal control group,all other groups were injected intraperitoneally with 5-FU(30 mg·kg^(-1))for seven consecutive days to establish a myocardial injury model.During the modeling process,the mice in the low-,medium-,and high-dose Xiao Xianxiongtang groups were administered the corresponding doses of Xiao Xianxiongtang by gavage.The normal control group and the model group were given equal volumes of normal saline for seven consecutive days.Left ventricular internal dimension at enddiastole(LVIDd),left ventricular internal dimension at end-systole(LVIDs),left ventricular ejection fraction(LVEF),and left ventricular fractional shortening(LVFS)were measured by color Doppler echocardiography.Histopathological changes in myocardial tissues were observed through hematoxylin-eosin(HE)and Masson staining.The protein expression of alpha-smooth muscle actin(α-SMA)was detected by immunohistochemistry.The levels of malondialdehyde(MDA)and superoxide dismutase(SOD)were detected by enzyme-linked immunosorbent assay(ELISA).The mRNA expression levels of HIF-1α and VEGF were detected by Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR).The protein expression levels of HIF-1α,VEGF,collagen Ⅰ(Col Ⅰ),and collagen Ⅲ(ColⅢ)were detected by Western blot.Result:Compared with the normal control group,the model group exhibited significantly increased LVIDd and LVIDs(P<0.01),decreased LVEF and LVFS(P<0.01),and pathological changes such as obvious disarray of myocardial tissue,
作者
叶倩倩
秦美洁
丁芮
方海雁
陈建
黄金玲
YE Qianqian;QIN Meijie;DING Rui;FANG Haiyan;CHEN Jian;HUANG Jinling(Anhui University of Chinese Medicine,Hefei 230012,China;Anhui Province Key Laboratory of Chinese Medicinal Formula,Hefei 230012,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2024年第16期1-9,共9页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81973844,81573864)。
