藏红花酸抗小鼠肝纤维化的实验研究
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摘要
目的:通过观察藏红花酸对肝纤维化小鼠肝组织病理学改变及p38MAPK蛋白表达的影响,探讨其抗肝纤维化作用及可能的机制。方法:36只6周龄雄性昆明小鼠随机分为3组:藏红花酸组、模型组、正常组。除正常对照组外,其余两组小鼠给予30%四氯化碳橄榄油溶液腹腔注射,首次注射5 m L/Kg,以后每次3 m L/Kg,每3日1次,连续12周。同时藏红花酸组给予藏红花酸5mg/Kg灌胃,每日1次,连续12周。实验过程中,观察各组小鼠的一般状态。12周末处死小鼠,肝组织切片行HE、Masson染色,显微镜下观察病理改变,免疫组化检测小鼠肝组织p38MAPK蛋白的表达。结果:实验过程中,模型组小鼠一般状态较差,体重增长缓慢;藏红花酸组小鼠一般状态尚可,体重较模型组增长明显(P<0.05)。肝组织HE、Masson染色结果显示:模型组小鼠肝纤维化程度较重(造模成功),与其相比,藏红花酸组小鼠肝纤维化程度有所改善,差异有统计学意义。与模型组相比,藏红花酸组肝组织p38MAPK表达显著下降(P<0.05)。结论:藏红花酸能有效减轻小鼠肝损伤及纤肝维化程度,其抗肝纤维化的机制可能与下调p38MAPK蛋白的表达有关。
Objective: To explore the effect and mechanism of crocetin on hepatic fibrosis by detecting the expression of p38 MAPK protetin and liver tissue pathological changes in mice with liver fibrosis. Methods: 36 six-week old male kunming mice were randomly divided into 3 groups: the crocetin group, model group and normal group. Except the mice of normal group, the others were given 30 % carbon tetrachloride olive oil solution by intraperitoneal injection to induce the mice models of hepatic fibrosis once every three days. And crocetin group mice were treated with crocetin by gavage everyday. During the experiment, the general condition of mice was recorded, such as the weight. After 12 weeks, the mice were sacrificed. Then their liver tissue were collected for study. The liver pathological changes of mice were observed by HE and Masson staining. The expression of p38 MAPK protein of mice liver tissue were detected by Immunohistochemistry(IHC). Results: During the experiment the general state of model group was gradually getting worse.Mice in crocetin group got better general state. Compared with the model group, the weight of crocetin group mice was increased(P <0.05). The difference was statistically significant(P<0.05). Liver tissue pathology results showed the degree of liver inflammation and fibrosis in model group was the heaviest and treatment with crocetin have a obvious improvement in the pathological changes. Compared with the model group, the expression of p38 MAPK protein of mice liver tissue in Crocetin group was reduced significantly(P <0.05).Conclusions: Crocetin could reduce the degree of liver injury and liver fibrosis of mice significantly, which might be related to the reduce the p38 MAPK protein expression.
Objective: To explore the effect and mechanism of crocetin on hepatic fibrosis by detecting the expression of p38 MAPK protetin and liver tissue pathological changes in mice with liver fibrosis. Methods: 36 six-week old male kunming mice were randomly divided into 3 groups: the crocetin group, model group and normal group. Except the mice of normal group, the others were given 30 % carbon tetrachloride olive oil solution by intraperitoneal injection to induce the mice models of hepatic fibrosis once every three days. And crocetin group mice were treated with crocetin by gavage everyday. During the experiment, the general condition of mice was recorded, such as the weight. After 12 weeks, the mice were sacrificed. Then their liver tissue were collected for study. The liver pathological changes of mice were observed by HE and Masson staining. The expression of p38 MAPK protein of mice liver tissue were detected by Immunohistochemistry(IHC). Results: During the experiment the general state of model group was gradually getting worse.Mice in crocetin group got better general state. Compared with the model group, the weight of crocetin group mice was increased(P <0.05). The difference was statistically significant(P<0.05). Liver tissue pathology results showed the degree of liver inflammation and fibrosis in model group was the heaviest and treatment with crocetin have a obvious improvement in the pathological changes. Compared with the model group, the expression of p38 MAPK protein of mice liver tissue in Crocetin group was reduced significantly(P <0.05).Conclusions: Crocetin could reduce the degree of liver injury and liver fibrosis of mice significantly, which might be related to the reduce the p38 MAPK protein expression.
引文
[1]Lai L,Chen Y,Tian X,et al.Artesunate alleviates hepatic fibrosis induced by multiple pathogenic factors and inflammation through the inhibition of LPS/TLR4/NF-κB signaling pathway in rats[J].European Journal of Pharmacology,2015,76(5):234-241
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[8]梅夏齐,汪云,王风秀,等.藏红花对肝纤维化大鼠肝脏组织中Caspase-3,Bcl-2表达的影响[J].现代生物医学进展,2016,16(23):4427-4430Mei Xia-qi,Wang Yun,Wang Feng-xiu,et al.The Influence of Saffron in Expression of Apoptosis Related Protein Caspase-3,Bcl-2in Rats with Liver Fibrosis[J].Progress in Modern Biomedicine,2016,16(23):4427-4430
[9]朱艳虹,陈真,钱之玉,等.西红花酸对乙醛诱导的肝星状细胞增殖和胶原合成的影响[J].中国临床药理学与治疗学,2013,18(8):841-847Zhu Yan-hong,Chen Zhen,Qian Zhi-yu,et al.The influence of crotein on hepatic stellate cells proliferation and collagen synthesis induced by acetaldehyde[J].Chinese Clinical Pharmacology and Therapeutics,2013,18(8):841-847
[10]中华医学会传染病与寄生虫病学分会.肝脏病学分会.病毒性肝炎防治方案[J].中华肝脏病杂志,2000,8(6):324-329The Infectious Diseases and Parasites Epidemiology branch,Hepatology branch of the Chinese Medical Association.The National Program for Prevention and Treatment of Viral Hepatitis[J].Chinese Journal of Hepatology,2000,8(6):324-329
[11]宁娜,韩建军.西红花酸药理作用的研究进展[J].海峡药学,2014,26(1):45-48Ning Na,Han Jian-jun.Research progress on pharmacological effects of Crocus sativus[J].Strait Pharmaceutical Journal,2014,26(1):45-48
[12]Rahaiee S,Moini S,Hashemi M,et al.Evaluation of antioxi dant activities of bioactive compounds and various extracts obtained from saffron(Crocus sativus L.):a review[J].Journal of food science and technology,2015,52(4):1-8
[13]He S Y,Qian Z Y,Wen N,et al.Influence of Crocetin on experimental atherosclerosis in hyperlipidamic-diet quails[J].European Journal of Pharmacology,2007,554(2-3):191-195
[14]胡慧,钱之玉,成文媛,等.西红花酸对心肌成纤维细胞增殖和胶原合成的影响[J].中国临床药理学与治疗学,2012,17(3):282-288Hu Hui,Qian Zhi-yu,Cheng Wen-yuan,et al.The influence of crotein on Cardiac fibroblasts proliferation and collagen synthesis[J].Chinese Clinical Pharmacology and Therapeutics,2012,17(3):282-288
[15]成文媛,钱之玉,杨丽娜,等.西红花酸在AngⅡ诱导的大鼠肺成纤维细胞中的作用[J].药学与临床研究,2012,20(2):103-108Cheng Wen-yuan,Qian Wen-yu,Yang Li-na,et al.Saffron acid role in rat lung fibroblasts induced by AngⅡ[J].Pharmaceutical and Clinical Research,2012,20(2):103-108
[16]Brichkina A,Nguyen N T,Baskar R,et al.Proline isomerisation as a novel regulatory mechanism for p38MAPK activation and functions[J].Cell Death&Differentiation,2016,1-10
[17]Zheng Na-na,Yue Ya-lun,Zheng Yong,et al.Hydrogen sulfide promotes proliferation of hepatocytes from hepatic fibrosis rats via p38MAPK signal pathway[J].World Chinese Journal of Digestology,2015(6):901-906
[18]吴文娟,杨妙芳,许小兵,等.p38MAPK在大鼠实验性肝纤维化发生中的表达及其意义[J].世界华人消化杂志,2008,16(34):3822-3827Wu Wen-juan,Yang Miao-fang,Xu Xiao-bing,et al.Expression and its locationof p38MAPK in CCl4-induced hepatic fibrosis in rats[J].China Journal of Traditional Chinese Medicine&Pharmacy,2008,16(34):3822-3827
[19]刘绍能,杨清高,潘澎,等.芪术颗粒对肝纤维化形成过程中p38MAPK信号传导通路的影响[J].中华中医药杂志,2014,29(7):2137-2140Liu Shao-neng,Yang Qing-gao,Pan Peng,et al.Study on effect of Qizhu Granule on p38MAPK signaling transduction pathways in the process of hepatic fi brosis[J].China Journal of Traditional Chinese Medicine&Pharmacy,2014,29(7):2137-2140
[20]郑人源,蒋明德,梅浙川,等.肝星状细胞增殖与p38丝裂原活化蛋白激酶信号传导通路的关系[J].中国组织工程研究,2011,15(20):3711-3714Zheng Ren-yuan,Jiang Ming-de,Mei Zhe-chuan,et al.Correlation between acetaldehyde-induced proliferation of hepatic stellate cells and p38mitogen-activated protein kinase signal transduction pathway[J].Journal of Clinical Rehabilitative Tissue Engineering Research,2011,15(20):3711-3714
[21]杨新疆,齐翠花,郑勇,等.SB203580对肝纤维化大鼠肝脏Ⅰ,Ⅲ型胶原蛋白表达的影响[J].世界华人消化杂志,2014,22(3):310-318Yang Xin-jiang,Qi Cui-hua,Zheng Yong,et al.SB203580 decreases collagenⅠand collagenⅢexpression in the liver of rats with experimental hepatic fibrosis[J].World Chinese Journal of Digestology,2014,22(3):310-318
[2]Shuiping Yu,Xueling Zhou,Bingzong Hou,et al.Protective effect of rosuvastatin treatment by regulating oxidized low-density lipoprotein expression in a rat model of liver fibrosis[J].Biomedical Reports,2016,5(3):311-316
[3]Liqian Zhu,Chen Yuan,Liyuan Huang,et al.The activation of p38MAPK and JNK pathways in bovine herpesvirus 1 infected MDBK cells[J].Veterinary Research,2016,47(1):1-8
[4]Jesús G C,Olga R,Cimas F J,et al.p38MAPK and Chemotherapy:We Always Need to Hear Both Sides of the Story[J].Frontiers in Cell&Developmental Biology,2016,4(69):1-8
[5]胡江宁,姚德中,章江生,等.西红花抗肿瘤作用的研究进展[J].安徽农业科学,2014,42(3):699-701,703Hu Jiang-ning,Yao De-zhong,Zhang Jiang-sheng,et al.Research Progress of Crocus sativus L.Antitumor Role[J].Journal of Anhui Agricultural Sciences,2014,42(3):699-701,703
[6]Omidi A,Riahinia N,Torbati M B M,et al.Hepatoprotective effect of Crocus sativus(saffron)petals extract against acetaminophen toxicity in male Wistar rats[J].Avicenna Journal of Phytomedicine,2014,4(5):330-336
[7]王强,钟丽华,于雷,等.藏红花对肝纤维化大鼠肝组织TGF-β表达的影响[J].现代生物医学进展,2012,12(17):3228-3231Wang Qiang,Zhong Li-hua,Yu Lei,et al.Effects of Zanghonghua on Expression of Transforming Growth Factors-β1 in Rats with Liver Fibrosis[J].Progress in Modern Biomedicine,2012,12(17):3228-3231
[8]梅夏齐,汪云,王风秀,等.藏红花对肝纤维化大鼠肝脏组织中Caspase-3,Bcl-2表达的影响[J].现代生物医学进展,2016,16(23):4427-4430Mei Xia-qi,Wang Yun,Wang Feng-xiu,et al.The Influence of Saffron in Expression of Apoptosis Related Protein Caspase-3,Bcl-2in Rats with Liver Fibrosis[J].Progress in Modern Biomedicine,2016,16(23):4427-4430
[9]朱艳虹,陈真,钱之玉,等.西红花酸对乙醛诱导的肝星状细胞增殖和胶原合成的影响[J].中国临床药理学与治疗学,2013,18(8):841-847Zhu Yan-hong,Chen Zhen,Qian Zhi-yu,et al.The influence of crotein on hepatic stellate cells proliferation and collagen synthesis induced by acetaldehyde[J].Chinese Clinical Pharmacology and Therapeutics,2013,18(8):841-847
[10]中华医学会传染病与寄生虫病学分会.肝脏病学分会.病毒性肝炎防治方案[J].中华肝脏病杂志,2000,8(6):324-329The Infectious Diseases and Parasites Epidemiology branch,Hepatology branch of the Chinese Medical Association.The National Program for Prevention and Treatment of Viral Hepatitis[J].Chinese Journal of Hepatology,2000,8(6):324-329
[11]宁娜,韩建军.西红花酸药理作用的研究进展[J].海峡药学,2014,26(1):45-48Ning Na,Han Jian-jun.Research progress on pharmacological effects of Crocus sativus[J].Strait Pharmaceutical Journal,2014,26(1):45-48
[12]Rahaiee S,Moini S,Hashemi M,et al.Evaluation of antioxi dant activities of bioactive compounds and various extracts obtained from saffron(Crocus sativus L.):a review[J].Journal of food science and technology,2015,52(4):1-8
[13]He S Y,Qian Z Y,Wen N,et al.Influence of Crocetin on experimental atherosclerosis in hyperlipidamic-diet quails[J].European Journal of Pharmacology,2007,554(2-3):191-195
[14]胡慧,钱之玉,成文媛,等.西红花酸对心肌成纤维细胞增殖和胶原合成的影响[J].中国临床药理学与治疗学,2012,17(3):282-288Hu Hui,Qian Zhi-yu,Cheng Wen-yuan,et al.The influence of crotein on Cardiac fibroblasts proliferation and collagen synthesis[J].Chinese Clinical Pharmacology and Therapeutics,2012,17(3):282-288
[15]成文媛,钱之玉,杨丽娜,等.西红花酸在AngⅡ诱导的大鼠肺成纤维细胞中的作用[J].药学与临床研究,2012,20(2):103-108Cheng Wen-yuan,Qian Wen-yu,Yang Li-na,et al.Saffron acid role in rat lung fibroblasts induced by AngⅡ[J].Pharmaceutical and Clinical Research,2012,20(2):103-108
[16]Brichkina A,Nguyen N T,Baskar R,et al.Proline isomerisation as a novel regulatory mechanism for p38MAPK activation and functions[J].Cell Death&Differentiation,2016,1-10
[17]Zheng Na-na,Yue Ya-lun,Zheng Yong,et al.Hydrogen sulfide promotes proliferation of hepatocytes from hepatic fibrosis rats via p38MAPK signal pathway[J].World Chinese Journal of Digestology,2015(6):901-906
[18]吴文娟,杨妙芳,许小兵,等.p38MAPK在大鼠实验性肝纤维化发生中的表达及其意义[J].世界华人消化杂志,2008,16(34):3822-3827Wu Wen-juan,Yang Miao-fang,Xu Xiao-bing,et al.Expression and its locationof p38MAPK in CCl4-induced hepatic fibrosis in rats[J].China Journal of Traditional Chinese Medicine&Pharmacy,2008,16(34):3822-3827
[19]刘绍能,杨清高,潘澎,等.芪术颗粒对肝纤维化形成过程中p38MAPK信号传导通路的影响[J].中华中医药杂志,2014,29(7):2137-2140Liu Shao-neng,Yang Qing-gao,Pan Peng,et al.Study on effect of Qizhu Granule on p38MAPK signaling transduction pathways in the process of hepatic fi brosis[J].China Journal of Traditional Chinese Medicine&Pharmacy,2014,29(7):2137-2140
[20]郑人源,蒋明德,梅浙川,等.肝星状细胞增殖与p38丝裂原活化蛋白激酶信号传导通路的关系[J].中国组织工程研究,2011,15(20):3711-3714Zheng Ren-yuan,Jiang Ming-de,Mei Zhe-chuan,et al.Correlation between acetaldehyde-induced proliferation of hepatic stellate cells and p38mitogen-activated protein kinase signal transduction pathway[J].Journal of Clinical Rehabilitative Tissue Engineering Research,2011,15(20):3711-3714
[21]杨新疆,齐翠花,郑勇,等.SB203580对肝纤维化大鼠肝脏Ⅰ,Ⅲ型胶原蛋白表达的影响[J].世界华人消化杂志,2014,22(3):310-318Yang Xin-jiang,Qi Cui-hua,Zheng Yong,et al.SB203580 decreases collagenⅠand collagenⅢexpression in the liver of rats with experimental hepatic fibrosis[J].World Chinese Journal of Digestology,2014,22(3):310-318