铁皮石斛多糖基于Notch信号通路抗大鼠肝纤维化作用及其超声评价  

Effect of Dendrobium officinale Polysaccharides in Reversing Hepatic Fibrosis in Rats via Notch Signaling Pathway and Ultrasound Evaluation

作　　者：许颖 罗继娜 杨柳青 田纤纤 成家茂 陈海燕 XU Ying;LUO Jina;YANG Liuqing;TIAN Qianqian;CHENG Jiamao;CHEN Haiyan(School of Clinical Medicine,Dali University,Dali 671003,China;Dali Bai Autonomous Prefecture People's Hospital,Dali 671000,China;School of Basic Medical Sciences,Dali University,Dali 671003,China;The First Affiliated Hospital of Dali University,Dali 671000,China)

机构地区：[1]大理大学临床医学院,云南大理671003 [2]大理白族自治州人民医院,云南大理671000 [3]大理大学基础医学院,云南大理671003 [4]大理大学第一附属医院,云南大理671000

出　　处：《中国实验方剂学杂志》2024年第21期70-77,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：云南省地方本科高校基础研究联合专项资金面上项目(202101BA070001~101);大理市科技项目(2022KBG007,2022KBG019);云南省教育厅科学研究基金项目(2023Y0962)。

摘　　要：目的:探讨铁皮石斛多糖(DOP)基于Notch信号通路逆转四氯化碳(CCl4)诱导大鼠肝纤维化(HF)的作用,并采用超声弹性成像技术评价药物疗效。方法:将56只雄性SD大鼠随机分成正常组、模型组、秋水仙碱组、扶正化瘀组、DOP低、中、高剂量组,每组8只。模型组及各给药组均皮下注射CCl4-橄榄油(2∶3)混合液,每3 d注射1次,共10周。造模6周后,各给药组分别给予秋水仙碱(1×10^(-4) g·kg^(-1))、扶正化瘀胶囊粉(0.45 g·kg^(-1))和低、中、高剂量DOP(0.05、0.1、0.2 g·kg^(-1))连续灌胃4周,每日1次。采用苏木素-伊红(HE)和马松(Masson)染色观察大鼠肝组织的病理形态学改变。采用全自动生化分析仪分析血清肝功能指标丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、总胆汁酸(TBA)和碱性磷酸酶(ALP)水平;采用酶联免疫吸附测定法(ELISA)检测血清肝纤四项指标透明质酸(HA)、层黏连蛋白(LN)、Ⅲ型前胶原(PC-Ⅲ)、Ⅳ型胶原(Col-Ⅳ)含量。分别采用实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白免疫印迹法(Western blot)检测大鼠肝组织中的α-平滑肌肌动蛋白(α-SMA)、Notch1、Jagged1、Hes1的mRNA和蛋白表达。采用声辐射力脉冲(ARFI)弹性成像技术获取大鼠肝脏的杨氏模量(YM),观察DOP用药前后YM的变化,并分析YM与肝纤四项、肝组织中α-SMA和Notch1信号通路相关因子蛋白的相关性。结果:与正常组比较,模型组大鼠的肝组织病理检查结果显示,肝组织肝细胞索排列紊乱,炎性细胞浸润明显,出现大量的脂肪空泡和纤维组织增生;大鼠的血清肝功能和肝纤四项表达均显著升高(P<0.01);大鼠肝组织中的α-SMA、Notch1、Jagged1、Hes1 mRNA和蛋白相对表达量均显著升高(P<0.01)。与模型组比较,各给药组大鼠肝组织的假小叶及汇管区胶原纤维沉积均有不同程度改善;除DOP低剂量组大鼠的血清ALT水平变化不明显外,各给药组均能显著降Objective:To investigate the effect of Dendrobium officinale polysaccharides(DOP)in reversing carbon tetrachloride(CCl4)-induced hepatic fibrosis(HF)in rats via the Notch signaling pathway and evaluate the therapeutic effect of DOP by ultrasound elastography.Method:Fifty-six male SD rats were randomized into normal,model,colchicine(1×10^(-4) g·kg^(-1)),Fuzheng Huayu powder(0.45 g·kg^(-1)),and low-,medium-,and high-dose(0.05,0.1,0.2 g·kg^(-1))DOP groups(n=8).The rats in the model group and each treatment group were injected subcutaneously with a mixture of CCl4-olive oil(2∶3)once every 3 days for 10 weeks.After 6 weeks of modelling,the rats were administrated with corresponding drugs once a day for 4 weeks.Hematoxylin-eosin(HE)staining and Masson staining were employed to observe the pathomorphological changes of the liver tissue.An automatic biochemical analyzer was used to measure the serum levels of alanine aminotransferase(ALT),aspartate transaminase(AST),alkaline phosphatase(ALP),and total bile acids(TBA).Enzyme-linked immunosorbent assay(ELISA)was employed to measure the serum levels of hyaluronic acid(HA),laminin(LN),typeⅢprecollagen(PC-Ⅲ),and typeⅣcollagen(Col-Ⅳ).The mRNA and protein levels ofα-smooth muscle actin(α-SMA),Notch1,Jagged1,and Hes1 in the liver tissue were determined by Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)and Western blot,respectively.The Young's modulus(YM)of the rat liver was measured by acoustic radiation force impulse(ARFI)elastography before and after treatment.Then,the correlations of YM with the serum levels of HA,LN,PC-Ⅲ,and Col-Ⅳand the protein levels ofα-SMA and Notch1 signaling pathway-related factors in the liver tissue were analyzed.Result:Compared with the normal group,the model group showed disordered arrangement of liver cell cords,obvious infiltration of inflammatory cells,appearance of a large number of fat vacuoles,and fibrous proliferation,elevated levels of ALT,AST,TBA,ALP,HA,LN,PC-Ⅲ,and Col-Ⅳin the serum,and

关 键 词：铁皮石斛多糖 肝纤维化 NOTCH信号通路 超声评价 杨氏模量 

分 类 号：R2-0[医药卫生—中医学] R22R285.5R289R33