复方片仔癀肝宝调控MicroRNA-155及IL-6、IL-10防治肝损伤的分子机制研究
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摘要
目的:基于MicroRNA-155(miR-155)靶点探讨复方片仔癀肝宝(GB)对大鼠急性肝损伤保护作用及下游白细胞介素-6(Interleukin-6,IL-6)/白细胞介素-10(Interleukin-10,IL-10)信号通路调节的分子机制。方法:SD大鼠60只按随机数字表法分为空白组、模型组、西利宾胺组(阳性对照组)、GB低剂量组、GB中剂量组和GB高剂量组,各实验组按照不同剂量灌胃给药0. 5ml/100g,连续7 d,每日1次。除空白组外其他各组末次给药1h后按2ml/kg给予50% CCL4花生油溶液腹腔注射造模,空白组腹腔注射2ml/kg花生油,检测血清中谷丙转氨酶(alanine aminotransferase,ALT)、谷草转氨酶(aspartate aminotransferase,AST)活性和IL-6、IL-10含量,观察肝组织病理学变化,Q-PCR法检测肝组织中IL-6、IL-10、miR-155表达。结果:GB能降低转氨酶的活性,显著抑制IL-6和miR-155的表达,升高IL-10的表达。结论:GB可能通过调节miR-155及其下游靶基因IL-6、IL-10的表达,降低肝损伤的炎症反应以达到抗肝损伤的作用。
Objective: To study the protective effect of Complex Prescription Pianzaihuang Ganbao( GB) on acute liver injury in rats and the molecular mechanism based on miR-155 targeting IL-6/IL-10 signaling pathway.Methods: 60 SD rats were randomly divided into normal group,model group,Silymarin group( positive control),GB low dose group,GB middle dose group,GB high dose group. Each experimental group was administered by gavage at 0. 5 ml/100 g,with respective doses,each day for 7 days. Except for the blank group,the other groups were injected with 50% CCL4 peanut oil solution2 ml/kg intraperitoneally 1 h after the last administration,and the blank group were intraperitoneally injected with 2 ml/kg peanut oil. Serum alanine aminotransferase( ALT),aspartate aminotransferase( AST) activity and IL-6,IL-10 content,Liver histopathology was observed. Q-PCR was used to explore the expression of IL-6,IL-10 and miR-155.Results: GB can reduce the activity of aminotransferases,significantly inhibited the expression of IL-6 and miR-155 and increased the expression of IL-10. Conclusion: GB may reduce the hepatic injury inflammatory response by regulating the expression of miR-155 and its downstream target genes IL-6 and IL-10 to achieve the effect of anti-hepatic injury.
Objective: To study the protective effect of Complex Prescription Pianzaihuang Ganbao( GB) on acute liver injury in rats and the molecular mechanism based on miR-155 targeting IL-6/IL-10 signaling pathway.Methods: 60 SD rats were randomly divided into normal group,model group,Silymarin group( positive control),GB low dose group,GB middle dose group,GB high dose group. Each experimental group was administered by gavage at 0. 5 ml/100 g,with respective doses,each day for 7 days. Except for the blank group,the other groups were injected with 50% CCL4 peanut oil solution2 ml/kg intraperitoneally 1 h after the last administration,and the blank group were intraperitoneally injected with 2 ml/kg peanut oil. Serum alanine aminotransferase( ALT),aspartate aminotransferase( AST) activity and IL-6,IL-10 content,Liver histopathology was observed. Q-PCR was used to explore the expression of IL-6,IL-10 and miR-155.Results: GB can reduce the activity of aminotransferases,significantly inhibited the expression of IL-6 and miR-155 and increased the expression of IL-10. Conclusion: GB may reduce the hepatic injury inflammatory response by regulating the expression of miR-155 and its downstream target genes IL-6 and IL-10 to achieve the effect of anti-hepatic injury.
引文
[1]彭飞,林丽,熊瑞芳.肝损伤机制的研究[J].科技资讯,2012,(33):236-237.
[2]陈志亮,赵锦燕,林珊,等.复方片仔癀肝宝对大鼠慢性酒精性肝损伤肝脏SREBP2和HMGCR表达的影响[J].时珍国医国药,2017,28(4):860-862.
[3]黄进明,赵锦燕,万芸,等.复方片仔癀肝宝对急性肝损伤肝的保护作用研究[J].世界中西医结合杂志,2015,10(9):1216-1218.
[4]陈达鑫,林珊,赵锦燕,等.复方片仔癀肝宝对酒精诱导的慢性肝损伤氧化应激的保护作用研究[J].实用中西医结合临床,2016,16(7):4-6.
[5]洪绯,赵锦燕,张泽修,等.复方片仔癀肝保片对大鼠四氯化碳诱导的慢性肝损伤的治疗作用[J].福建中医药,2014,45(5):53-54.
[6]陈达鑫,林珊,赵锦燕,等.复方片仔癀肝宝对酒精性肝病的抗炎作用[J].福建中医药,2016,47(5):6-8.
[7]胡冬梅,陆杨,房敏峰,等.特女贞苷对四氯化碳致小鼠急性肝损伤的保护作用[J].中国药理学通报,2016,32(9):1260-1263.
[8]张力,万敬员,段红,等.白介素-10表达上调介导脂氧素在急性肝损伤中的保护效应[J].医学分子生物学杂志,2008,5(3):225-228.
[9]刘庆春,张峰,黄川锋,等.丁酸钠对内毒素性肝损伤小鼠炎症因子表达的影响[J].中国临床研究,2016,29(7):954-956.
[10]王越,沈连忠,李波.临床研究中肝损伤的临床病理指标的选择及意义[J].中国药事,2009,23(8):813-825.
[11] POGRIBNY IP,STARLARD-DAVERPORT A,TRYNDYAK VP,et al. Difference in expression of hepatic microRNAs miR-29c,miR-34a,miR-155,and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice[J].Lab Invest,2010,90(10):1437-1446.
[12] YUAN K,ZHANG X,LV L,et al.Fine-tuning the expression of microRNA-155 controls acetaminophen-induced liver inflammation[J]. International Immunopharmacology,2016,40(11):339-346.
[13] WAN C,HAN R,LIU L,et al. Role of miR-155 in fluorooctane sulfonate-induced oxidative hepaticdamage via the Nrf2-dependent pathway[J]. Toxicology and Applied Pharmcology,2016,295(3):85-93.
[14] ARRANZ A,DOXAKI C,VERGADI E,er al. Aktl and Akt2protein kinases differentially contribute to macrophage polarization[J].Proceedings of the National Academy of Sciences,2012,109(24):9517-9522.
[15] CARDOSO A L,GUEDES J R,PEREIRA D A,er al. miR-155modulates microglia mediated inmmune response by downregulating SOCSI and promoting cytokine and nitric oxide production[J].Immunology,2012,135(1):73-88.
[16]王铖芸,张凡,侯敏,等.microRNA在小鼠血吸虫病及吡喹酮治疗中的表达特征[J].中国血吸虫病防治杂志,2014,26(2):165-168.
[17] YANG LL,LIU JQ,BAI XZ,et al. Acute down regulation of miR155 at wound sites leads to a reduced fibrosis through at tenuating inflammatory response[J]. Bionchem biophys Res Commun,2014,453(1):153-159.
[18]张力,万敬员,段红,等.白介素-10表达上调介导脂氧素在急性肝损伤中的保护效应[J].医学分子生物学杂志,2008,5(3):225-228.
[19]刘庆春,张峰,黄川锋,等.丁酸钠对内毒素性肝损伤小鼠炎症因子表达的影响[J].中国临床研究,2016,29(7):954-956.
[2]陈志亮,赵锦燕,林珊,等.复方片仔癀肝宝对大鼠慢性酒精性肝损伤肝脏SREBP2和HMGCR表达的影响[J].时珍国医国药,2017,28(4):860-862.
[3]黄进明,赵锦燕,万芸,等.复方片仔癀肝宝对急性肝损伤肝的保护作用研究[J].世界中西医结合杂志,2015,10(9):1216-1218.
[4]陈达鑫,林珊,赵锦燕,等.复方片仔癀肝宝对酒精诱导的慢性肝损伤氧化应激的保护作用研究[J].实用中西医结合临床,2016,16(7):4-6.
[5]洪绯,赵锦燕,张泽修,等.复方片仔癀肝保片对大鼠四氯化碳诱导的慢性肝损伤的治疗作用[J].福建中医药,2014,45(5):53-54.
[6]陈达鑫,林珊,赵锦燕,等.复方片仔癀肝宝对酒精性肝病的抗炎作用[J].福建中医药,2016,47(5):6-8.
[7]胡冬梅,陆杨,房敏峰,等.特女贞苷对四氯化碳致小鼠急性肝损伤的保护作用[J].中国药理学通报,2016,32(9):1260-1263.
[8]张力,万敬员,段红,等.白介素-10表达上调介导脂氧素在急性肝损伤中的保护效应[J].医学分子生物学杂志,2008,5(3):225-228.
[9]刘庆春,张峰,黄川锋,等.丁酸钠对内毒素性肝损伤小鼠炎症因子表达的影响[J].中国临床研究,2016,29(7):954-956.
[10]王越,沈连忠,李波.临床研究中肝损伤的临床病理指标的选择及意义[J].中国药事,2009,23(8):813-825.
[11] POGRIBNY IP,STARLARD-DAVERPORT A,TRYNDYAK VP,et al. Difference in expression of hepatic microRNAs miR-29c,miR-34a,miR-155,and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice[J].Lab Invest,2010,90(10):1437-1446.
[12] YUAN K,ZHANG X,LV L,et al.Fine-tuning the expression of microRNA-155 controls acetaminophen-induced liver inflammation[J]. International Immunopharmacology,2016,40(11):339-346.
[13] WAN C,HAN R,LIU L,et al. Role of miR-155 in fluorooctane sulfonate-induced oxidative hepaticdamage via the Nrf2-dependent pathway[J]. Toxicology and Applied Pharmcology,2016,295(3):85-93.
[14] ARRANZ A,DOXAKI C,VERGADI E,er al. Aktl and Akt2protein kinases differentially contribute to macrophage polarization[J].Proceedings of the National Academy of Sciences,2012,109(24):9517-9522.
[15] CARDOSO A L,GUEDES J R,PEREIRA D A,er al. miR-155modulates microglia mediated inmmune response by downregulating SOCSI and promoting cytokine and nitric oxide production[J].Immunology,2012,135(1):73-88.
[16]王铖芸,张凡,侯敏,等.microRNA在小鼠血吸虫病及吡喹酮治疗中的表达特征[J].中国血吸虫病防治杂志,2014,26(2):165-168.
[17] YANG LL,LIU JQ,BAI XZ,et al. Acute down regulation of miR155 at wound sites leads to a reduced fibrosis through at tenuating inflammatory response[J]. Bionchem biophys Res Commun,2014,453(1):153-159.
[18]张力,万敬员,段红,等.白介素-10表达上调介导脂氧素在急性肝损伤中的保护效应[J].医学分子生物学杂志,2008,5(3):225-228.
[19]刘庆春,张峰,黄川锋,等.丁酸钠对内毒素性肝损伤小鼠炎症因子表达的影响[J].中国临床研究,2016,29(7):954-956.