裂果薯总皂苷抗大鼠肝纤维化作用及其机制
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摘要
目的研究裂果薯总皂苷(SFSP)对四氯化碳(CCl_4)诱导的肝纤维化大鼠的作用及其机制。方法将♂SD大鼠随机分为5组:正常对照组、模型组、秋水仙碱组(Col)、SFSP低剂量组(SFSP-L)、SFSP高剂量组(SFSP-H)。连续给药4周后,检测各组大鼠ALT、AST和羟脯氨酸的含量;ELISA法检测肝纤维化标志物;HE染色观察肝组织病理学改变;Masson染色观察肝纤维化程度;透射电镜观察肝超微结构变化;qPCR法检测α-SMA、TGF-β1、Smad4、Smad7 mRNA的表达情况。结果肝纤维化模型组AST、ALT及CⅣ、PCⅢ、LN、HA、Hyp反映肝损伤和纤维化进程的指标,均明显升高(P<0.05),而SFSP-H组和Col组明显降低上述指标含量,HE、Masson染色结果与之一致;SFSP明显抑制TGF-β1、Smad4、α-SMA mRNA的表达(P<0.01),升高Smad7的表达(P<0.01)。结论 SFSP通过抑制TGF-β1/Smad信号通路,明显抑制大鼠肝纤维化程度。
Aim To study the effect of total saponins of Schizocapsa plantaginea Hance(SFSP) on hepatic fibrosis in rats induced by CCl_4 and to investigate the molecular mechanisms of these effects. Methods ♂SD rats were randomly divided into five groups: normal control group, model group, colchicine(Col) as positive control group, low-dose SFSP group(SFSP-L), and high-dose SFSP group(SFSP-H). After four weeks of continuous administration, serum AST, ALT and hydroxyproline(Hyp) were measured by biochemical detection method. The four indicators of liver fibrosis(CIV, PCⅢ, LN, HA) in serum were measured by enzyme linked immunosorbent assay. The location of liver fibrous tissues was observed by Masson staining. Liver sections were stained with HE, and observed for pathologic changes. Liver ultrastructural changes were observed using a transmission electron microscope. The expressions of mRNA of α-smooth muscle actin(α-SMA), transforming growth factor β1(TGF-β1), Smad4 and Smad7 in liver were detected by qPCR. Results In the liver fibrosis model group, the contents of AST, ALT and CIV, PCⅢ, LN, HA, Hyp reflecting liver injury and fibrosis significantly increased(P<0.05), while in SFSP-H group and Col group the contents of the above indicators were significantly reduced, and the pathological results of liver tissues were consistent with it. SFSP significantly inhibited the expression of TGF-β1, Smad4 and α-SMA mRNA(P<0.01), and increased the expression of Smad7 as well(P<0.01). Conclusion SFSP has an antifibrogenic effect through down-regulating the TGF-β1/Smad signaling pathway in rats.
Aim To study the effect of total saponins of Schizocapsa plantaginea Hance(SFSP) on hepatic fibrosis in rats induced by CCl_4 and to investigate the molecular mechanisms of these effects. Methods ♂SD rats were randomly divided into five groups: normal control group, model group, colchicine(Col) as positive control group, low-dose SFSP group(SFSP-L), and high-dose SFSP group(SFSP-H). After four weeks of continuous administration, serum AST, ALT and hydroxyproline(Hyp) were measured by biochemical detection method. The four indicators of liver fibrosis(CIV, PCⅢ, LN, HA) in serum were measured by enzyme linked immunosorbent assay. The location of liver fibrous tissues was observed by Masson staining. Liver sections were stained with HE, and observed for pathologic changes. Liver ultrastructural changes were observed using a transmission electron microscope. The expressions of mRNA of α-smooth muscle actin(α-SMA), transforming growth factor β1(TGF-β1), Smad4 and Smad7 in liver were detected by qPCR. Results In the liver fibrosis model group, the contents of AST, ALT and CIV, PCⅢ, LN, HA, Hyp reflecting liver injury and fibrosis significantly increased(P<0.05), while in SFSP-H group and Col group the contents of the above indicators were significantly reduced, and the pathological results of liver tissues were consistent with it. SFSP significantly inhibited the expression of TGF-β1, Smad4 and α-SMA mRNA(P<0.01), and increased the expression of Smad7 as well(P<0.01). Conclusion SFSP has an antifibrogenic effect through down-regulating the TGF-β1/Smad signaling pathway in rats.
引文
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[2] Dong C,Gongora R,Sosulski M L,et al.Regulation of transforming growth factor-beta1 (TGF-β1)-induced pro-fibrotic activities by circadian clock gene Bmal1[J].Respir Res,2016,17:4.
[3] Fan X,Zhang Q,Li S,et al.Attenuation of CCl4-induced hepatic fibrosis in mice by vaccinating against TGF-β1[J].PLoS One,2013,8(12):e82190.
[4] 何伟,宋莎莎,袁平凡,等.雷公藤红素对二乙基亚硝胺诱导的大鼠肝纤维化的治疗作用及机制[J].中国药理学通报,2013,29(4):519-24.[4] He W,Song S S,Yuan P F,et al.Therapeutic effects of celastrol on DEN induced liver fibrosis rats and its mechanisms[J].Chin Pharmacol Bull,2013,29(4):519-24.
[5] Wang Y,Wang R,Wang Y,et al.Ginkgo Biloba extract mitigates liver fibrosis and apoptosis by regulating p38 MAPK,NF-κB/ⅠκBα,and Bcl-2/Bax signaling[J].Drug Des Devel Ther,2015,9:6303-17.
[6] 孙悦文.广西特色中草药靶向抗肿瘤筛选及裂果薯皂苷的分离与抗肿瘤作用研究[D].南宁:广西医科大学,2013.[6] Sun Y W.Targeted screening anti-cancer from herbal medicine in Guangxi and the isolation and anti cancer research of steroidal saponins from Schizocapsa Plantaginea Hance[D].Nanning:Guangxi Medical University,2013.
[7] 王金妮.利用抗肿瘤药靶—酪氨酸激酶从广西特色中草药中筛选和研究选择性抗肿瘤药[D].南宁:广西医科大学,2012.[7] Wang J N .Screenning and research selective anti-hepatoma drugs from the traditional Chuang and Yao medicine in Guangxi by tyrosine kinase,angiogenesis as antitumor drug targets [D].Nanning:Guangxi Medical University,2012.
[8] 欧明春,孙悦文,刘布鸣,等.裂果薯醇提物对人肝癌裸鼠移植瘤生长与血管生成的影响[J].中国实验方剂学杂志,2015,21(8):106-10.[8] Ou M C,Sun Y W,Liu B M,et al.Effects of ethanol extract from Schizocapsa plantaginea on growth and angiogenesis of human hepatocellular carcinoma xenografts in nude mice [J].Chin J Exp Tradit Med Form,2015,21(8):106-10.
[9] Yu J,Wu C W,Chu E S,et al.Elucidation of the role of COX-2 in liver fibrogenesis using transgenic mice[J].Mol Cell Biol Res Commun,2008,372(4):571-7.
[10] 洪燕,韩燕全,罗欢,等.重楼皂苷对肝纤维化大鼠纤维化标志物的影响及其相关性分析[J].山西中医学院学报,2014,15(6):20-2,67.[10] Hong Y,Han Y Q,Luo H,et al.Effect of Chonglou saponin on markers of liver fibrosis of hepatic fibrosis rats and correlation analysis[J].J Shanxi Coll Tradit Chin Med,2014,15(6):20-2,67.
[11] 樊威.重楼皂苷及其复方抗肝癌和安全性评价研究[D].天津:天津科技大学,2015.[11] Fan W.Study on anti-hepatocarcinoma and safety evaluation of Rhizoma Paridis saponins and its compound [D].Tianjin:Tianjin University of Science & Technology,2015.
[12] 禤传凤,罗伟生,陈国忠,等.中药活性成分干预肝纤维化分子信号通路的研究进展[J].中国药理学通报,2017,33(12):1638-41.[12] Xuan C F ,Luo W S,Chen G Z,et al.Study of active ingredients of Chinese medicine interfering with molecular signaling pathway of liver fibrosis[J].Chin Pharmacol Bull,2017,33(12):1638-41.
[13] Bian E,Huang C,Wang H,et al.Repression of Smad7 mediated by Dnmt1 determines hepatic stellate cell activation and liver fibrosis in rats[J].Toxicol Lett,2014,224(2):175-85.
[14] Wu M S,Liao C W,Du W Y,et al.Enhanced expression of transforming growth factor-beta 1 in inflammatory cells,alpha-smooth muscle actin in stellate cells,and collagen accumulation in experimental granulomatous hepatitis caused by Toxocara canis in mice[J].Acta Trop,2008,105(3):260-8.