Hh信号通路因子Shh、Ptch1和Gli1在大黄蛰虫丸抗大鼠肝纤维化中的表达及调节作用
|
摘要
目的探讨大黄蛰虫丸(DZP)对CCl4诱导肝纤维化大鼠肝脏Hedgehog(Hh)信号通路因子Shh、Gli1、Ptch1的影响及其机制。方法选SD雄性大鼠30只,体质量180～200 g,随机分为对照组(N组)、模型组(M组)和灌药组(D组)。后2组行CCl_4诱导肝纤维化造模,8周后D组、C组大鼠分别给予DZP药液灌胃,NG组大鼠给予等量生理盐水。连续灌胃4周后,断头处死各组大鼠,摘取肝脏,应用HE和Masson染色法检测各组别大鼠肝组织纤维化程度;应用Western blot和RT-PCR法分别检测各组大鼠肝组织Hh信号通路因子Shh、Ptch1、Gli1的蛋白及基因表达水平,用Quantity One图像分析软件进行条带光密度分析,计算光密度值(A值)。结果 M组大鼠肝纤维化明显,而D组大鼠肝的纤维化程度显著改善。Hh信号通路因子Shh、Ptch1、Gli1的基因和蛋白表达水平检测结果显示:与N组相比,M组大鼠肝组织中三者的A值均显著上调(P<0.05);而与M组相比,D组大鼠肝组织中三者的A出现显著下调(P<0.05),数据表明大鼠肝纤维化造模成功后,给予DZP干扰,后者可能通过抑制Shh、Gli1、Ptch1的基因和蛋白表达,从而抑制了肝纤维化过程中异常激活的Hh信号通路。结论 DZP可能通过抑制异常激活的Hh信号通路来逆转大鼠肝纤维化。
Objective To investigate the effect and its mechanism of Dahuang zhechong pill(DZP)on Shh,Gli1,Ptch1 of Hedgehog(Hh)signal pathway factors in the hepatic tissues ofliver fibrosisSD rats of weight 150-200 g were randomly divided into the normal control group(N group),model group(M group)and drug group(D group),the ones of latter two groups were used to establish theliver fibrosis model.After 8 weeks,the rats in DG were given DZP liquid medicine by gavage,while the ones of other two groups only the normal saline.After feeding for 4 weeks,the rats in each group were sacrificed by breaking head,and their livers were cut out partly for detecting the fibrosis degree of the rat liver tissues in each group by HE and Masson staining methods;the other liver tissues were used to measure the expression of Shh,Ptch1 and Gli1 proteins and genes of Hh signaling factors separately by Western blot and RT-PCR methods,calculating the values of A with image analysis software Quantityliver fibrosis of rats in M group was obvious,while the degree of hepatic fibrosis in D group rats was significantly improved.The gene and protein tests of Shh,Gli1 and Ptch1 of Hh signaling pathway factors showed that:Avalues of the three factors in liver tissues of rats in M group were significantly increased than in N group(P<0.05),and reduced in D group than in M group significantly(P<0.05).These data show that DZP can probably inhibit the abnormal activation of Hh signaling pathway in hepatic fibrogenesis by controlling the gene and protein expression of Shh,Ptch1 andmay reverse the liver fibrosis in rats by inhibiting the abnormal activation of Hh signaling pathway.
Objective To investigate the effect and its mechanism of Dahuang zhechong pill(DZP)on Shh,Gli1,Ptch1 of Hedgehog(Hh)signal pathway factors in the hepatic tissues ofliver fibrosisSD rats of weight 150-200 g were randomly divided into the normal control group(N group),model group(M group)and drug group(D group),the ones of latter two groups were used to establish theliver fibrosis model.After 8 weeks,the rats in DG were given DZP liquid medicine by gavage,while the ones of other two groups only the normal saline.After feeding for 4 weeks,the rats in each group were sacrificed by breaking head,and their livers were cut out partly for detecting the fibrosis degree of the rat liver tissues in each group by HE and Masson staining methods;the other liver tissues were used to measure the expression of Shh,Ptch1 and Gli1 proteins and genes of Hh signaling factors separately by Western blot and RT-PCR methods,calculating the values of A with image analysis software Quantityliver fibrosis of rats in M group was obvious,while the degree of hepatic fibrosis in D group rats was significantly improved.The gene and protein tests of Shh,Gli1 and Ptch1 of Hh signaling pathway factors showed that:Avalues of the three factors in liver tissues of rats in M group were significantly increased than in N group(P<0.05),and reduced in D group than in M group significantly(P<0.05).These data show that DZP can probably inhibit the abnormal activation of Hh signaling pathway in hepatic fibrogenesis by controlling the gene and protein expression of Shh,Ptch1 andmay reverse the liver fibrosis in rats by inhibiting the abnormal activation of Hh signaling pathway.
引文
[1]Jung YK,Yim HJ.Reversal of liver cirrhosis:current evidence and expectations[J].Korean J Intern Med,2017,32(2):213-228.
[2]Shen X,Peng Y,Li H.The injury-related activation of hedgehog signaling pathway modulates the repairassociated inflammation in liver fibrosis[J].Fron Immunol,2017,8:1450.
[3]Liu HL,Gu DS,Xie JW.Clinical implications of hedgehog signaling pathway inhibitors[J].Chin JCancer,2011,30(1):13-26.
[4]Robbins DJ,Fei DL,Riobo NA.The Hedgehog signal transduction network[J].Sci Signal,2012,5(246):re6.
[5]成家茂,潘志恒,谢瑶,等.大黄蛰虫丸经旁分泌途径对早期肝纤维化的干预作用[J].时珍国医国药,2010,21(2):296-299.
[6]潘志恒,成家茂,李永伟,等.大黄蛰虫丸对大鼠肝星状细胞活化增殖的影响[J].中山大学学报(医学科学版),2009,30(3):250-254.
[7]王德莉.大黄蛰虫丸治疗乙肝纤维化疗效的系统评价[D].成都中医药大学,2009.
[8]Li R,Cai L,Hu CM,et al.Expression of hedgehog signal pathway in articular cartilage is associated with the severity of cartilage damage in rats with adjuvant-induced arthritis[J].J Inflamm(Lond),2015,12(1):24.
[9]Skoda AM,Simovic D,Karin V,et al.The role of the Hedgehog signaling pathway in cancer:A comprehensive review[J].Bosn J Basic Med Sci,2018,18(1):8-20.
[10]Hu L,Lin X,Lu H,et al.An overview of hedgehog signaling in fibrosis[J].Mol Pharmacol,2015,87(2):174-82.
[11]罗秀芳,陈菊屏.Hedgehog信号通路与纤维化疾病的研究进展[J].现代临床医学,2016,42(2):99-101.
[12]孙艳.2,5-己二酮致人卵巢颗粒细胞凋亡及Sonichedgehog信号通路调控机制的研究[D].福建医科大学,2012:95-96.
[13]Hyun J,Jung Y.MicroRNAs in liver fibrosis:Focusing on the interaction with hedgehog signaling[J].World J Gastroenterol,2016,22(29):6652-6662.
[14]於琳,刘靳波.肿瘤微小RNA与hedgehog信号通路的研究进展[J].临床检验杂志,2014,32(8):597-600.
[15]Kalderon D.Transducing the hedgehog signal[J].Cell,2000,103(3):371-374.
[2]Shen X,Peng Y,Li H.The injury-related activation of hedgehog signaling pathway modulates the repairassociated inflammation in liver fibrosis[J].Fron Immunol,2017,8:1450.
[3]Liu HL,Gu DS,Xie JW.Clinical implications of hedgehog signaling pathway inhibitors[J].Chin JCancer,2011,30(1):13-26.
[4]Robbins DJ,Fei DL,Riobo NA.The Hedgehog signal transduction network[J].Sci Signal,2012,5(246):re6.
[5]成家茂,潘志恒,谢瑶,等.大黄蛰虫丸经旁分泌途径对早期肝纤维化的干预作用[J].时珍国医国药,2010,21(2):296-299.
[6]潘志恒,成家茂,李永伟,等.大黄蛰虫丸对大鼠肝星状细胞活化增殖的影响[J].中山大学学报(医学科学版),2009,30(3):250-254.
[7]王德莉.大黄蛰虫丸治疗乙肝纤维化疗效的系统评价[D].成都中医药大学,2009.
[8]Li R,Cai L,Hu CM,et al.Expression of hedgehog signal pathway in articular cartilage is associated with the severity of cartilage damage in rats with adjuvant-induced arthritis[J].J Inflamm(Lond),2015,12(1):24.
[9]Skoda AM,Simovic D,Karin V,et al.The role of the Hedgehog signaling pathway in cancer:A comprehensive review[J].Bosn J Basic Med Sci,2018,18(1):8-20.
[10]Hu L,Lin X,Lu H,et al.An overview of hedgehog signaling in fibrosis[J].Mol Pharmacol,2015,87(2):174-82.
[11]罗秀芳,陈菊屏.Hedgehog信号通路与纤维化疾病的研究进展[J].现代临床医学,2016,42(2):99-101.
[12]孙艳.2,5-己二酮致人卵巢颗粒细胞凋亡及Sonichedgehog信号通路调控机制的研究[D].福建医科大学,2012:95-96.
[13]Hyun J,Jung Y.MicroRNAs in liver fibrosis:Focusing on the interaction with hedgehog signaling[J].World J Gastroenterol,2016,22(29):6652-6662.
[14]於琳,刘靳波.肿瘤微小RNA与hedgehog信号通路的研究进展[J].临床检验杂志,2014,32(8):597-600.
[15]Kalderon D.Transducing the hedgehog signal[J].Cell,2000,103(3):371-374.