前列腺素E2对急性肝衰竭小鼠肝脏的保护作用及促肝再生作用的研究

  本文关键词：前列腺素E_2对急性肝衰竭小鼠肝脏的保护作用及促肝再生作用的研究，由笔耕文化传播整理发布。

        第一部分前列腺素E2对急性肝衰竭小鼠肝脏的保护作用研究目的：本研究通过建立对乙酰氨基酚(acetaminophen, APAP)诱导的小鼠急性肝衰竭模型,研究前列腺素E2(prostaglandin E2, PGE2)对APAP导致的急性肝衰竭小鼠肝脏的保护作用。方法：健康8周龄ICR小鼠,随机分为模型对照组、PGE2治疗组和正常对照组。正常对照组小鼠给予生理盐水腹腔注射和无菌PBS皮下注射；模型对照组小鼠APAP腹腔注射造模后给予无菌PBS皮下注射治疗；PGE2治疗组小鼠APAP腹腔注射造模后给予dmPGE2皮下注射治疗。各组小鼠造模后第12小时、24小时、48h处死采血检测血清谷丙转氨酶(alanine aminotransferase, ALT)和总胆红素(total bilirubin, TBIL)浓度,取肝脏组织进行病理检查并对肝组织损伤程度进行分级。结果：模型对照组小鼠血清ALT在造模后12h、24h、48h均显著高于正常对照组(P<0.05)；造模后12h、24h PGE2治疗组小鼠血清ALT低于同时间点模型对照组(P<0.05),48h两组差别未达到统计学显著性(P>0.05)。造模后12h模型对照组小鼠血清TBIL较正常对照组升高(P<0.05), PGE2治疗组较模型对照组降低(P<0.05)；24h、48h各处理组间无明显差异(P<0.05)。肝组织病理学检查显示模型对照组小鼠在造模后12h已经出现了明显的肝脏损害表现,12h、24h、48h肝损伤分级分别为4.0±0.7级、3.8±0.8级、2.4±1.1级。PGE2治疗组造模后12h、24h、48h肝损伤平均分级分别为1.4±0.5级、2.2±0.8级、1.2±0.4级,与模型对照组相比明显减轻(P<0.05)。结论：PGE2不但能减轻APAP所造成的肝损伤,还可以加快肝损伤的恢复,具有开发为治疗急性肝衰竭的有效药物的潜力。第二部分前列腺素E2对急性肝衰竭小鼠肝再生促进作用及其与HGF、Wnt2及β-catenin关系的研究目的：通过观察APAP诱导的肝衰竭小鼠肝脏再生情况及检测HGF、Wnt2及β-catenin在肝脏的表达水平,研究PGE2对肝衰竭小鼠肝再生的影响,及HGF、 Wnt2及β-catenin在PGE2调节肝再生过程中所发挥的作用。方法：取各处理组小鼠造模后12h、24h、48h肝组织石蜡切片进行增殖细胞核抗原(proliferating cell nuclear antigen, PCNA)免疫组化染色,以检测肝细胞分裂增殖情况；用RT-PCR及Western Blot检测肝脏组织中HGF、Wnt、β-catenin mRNA及蛋白水平的改变。结果：在造模后12h、24h,模型对照组小鼠肝脏PCNA阳性率低于正常对照组,但差别未达到统计学显著性(P>0.05),48h模型对照组小鼠肝脏PCNA阳性率高于正常对照组(15.4±4.8%vs10.0±1.9%,P<0.05)；同时间点相比,PGE2治疗组小鼠肝脏PCNA阳性率均高于正常对照组和模型对照组,其中24h、48h差别达到统计学显著性(P<0.05)。各时间点模型对照组小鼠与正常对照组小鼠相比,肝脏HGF和β-catenin mRNA表达无显著差异(P>0.05); PGE2治疗组小鼠肝脏HGF和β-catenin mRNA表达较模型对照组升高(P<0.05)。造模后12h PGE2治疗组小鼠肝脏Wnt2mRNA表达高于模型对照组(P<0.05),但在24h、48h两组相比无明显差别(P>0.05)。12h、24h PGE2治疗组小鼠HGF蛋白表达水平高于模型对照组,各组小鼠肝脏β-catenin、Wnt2蛋白水平表达改变不明显。结论：PGE2具有促进APAP导致的急性肝衰竭小鼠肝脏再生的作用,其促肝再生效应可能与HGF和β-catenin作用相关。

    Part ⅠThe Role of Prostaglandin E2in Alleviating Liver Indury in Acute Liver Failure in MiceAims:To investigate the role of prostaglandin E2(PGE2) in alleviating acute liver injury induced by acetaminophen (APAP) in mice.Methods:Eight-week ICR mice were randomly divided into three groups:(1) the normal control group:normal mice treated with saline;(2) the liver failure group: APAP-induced liver failure mice treated by phosphate buffer saline; and (3) PGE2-treated group:APAP-induced liver failure mice treated by dmPGE2; The mice were killed and samples were obtained at12hours,24hours,48hours after APAP administration, respectively. Serum alanine aminotransferase (ALT) and total bilirubin (TBIL) were measured and sections of liver were prepared for histological examination.Results:The level of serum ALT of the liver failure group mice was elevated at12h,24h,48h after APAP administration comparing to normal mice; and it was lower in PGE2treated mice at12h and24h comparing to the live failure group (P<0.05), while there was no significant difference at48h (P>0.05). The level of serum TBIL of liver failure mice was elevated at12h after APAP administration comparing to normal mice whereas down-regulated in mice of PGE2-treated group(P<0.05); however, no significant difference were found among the three groups at24h and48h (P>0.05). Severe liver damage was observed at12h after APAP administration by histological examination, and the degree of liver injury of the mice in liver failure group at12h,24h,48h were4.0±0.7,3.8±0.8,2.4±1.1, respectively; while he degree of liver injury of the mice in PGE2-treated group at12h,24h,48h were1.4±0.5,2.2±0.8,1.2±0.4, respectively. The PGE2group showed slighter liver injury than the liver failure group (P<0.05).Conclusions:PGE2can alleviate APAP-induced acute liver injury in mice and facilitate the recovery of the liver. It might be a potential medication for treating liver failure. Part ⅡThe Effect of Prostaglandin E2in Liver Regeneration of Acute Liver Failure in Mice and the Correlationship with HGF, Wnt2and β-cateninAims:To investigate the effect of PGE2in enhancing liver regeneration in mice with APAP-induced acute liver failure and the role of HGF, Wnt2and β-catenin in regulation of liver regeneration by PGE2.Methods:Proliferating cell nuclear antigen (PCNA) which indicating the level of liver regeneration was evaluate by immunohistochemistry on sections of paraffin-embedded liver tissue. The expression of HGF, Wnt2and β-catenin was evaluated by reverse transcription-polymerase chain reaction and western blot.Results:The rate of PCNA-positive cells in liver of liver failure mice group was down-regulated at12h and24h after APAP administration comparing to normal group, but the difference did not reach the statistic significance (P>0.05); and it was up-regulated at48h (15.4±4.8%vs10.0±1.9%, P<0.05). Treatment of PGE2up-regulated the rate of PCNA-positive cells at24h and48after APAP administration respectively, comparing to other groups (P<0.05). The expression of HGF and (3-catenin mRNA was up-regulated in mice of PGE2-treated group comparing to liver failure group(P<0.05), while there was no significant difference between normal group and liver failure group(P>0.05). The expression of Wnt2mRNA was up-regulated in PGE2-treated group comparing to liver failure group at12h after APAP-administration(P<0.05). The level of β-catenin and Wnt2protein showed no changed by western blot; while the level of HGF protein was up-regulated in mice of PGE2-treated group comparing to liver failure group at12h and24h, respectively.Conclusions:PGE2was capable of enhancing liver regeneration in mice with APAP-induced acute liver failure and this effect might correlate to HGF and β-catenin.

前列腺素E_2对急性肝衰竭小鼠肝脏的保护作用及促肝再生作用的研究

缩略语表4-5中文摘要5-7Abstract7-8引言9-14    参考文献10-14第一部分 前列腺素E_2对急性肝衰竭小鼠肝脏的保护作用研究14-29    前言14    材料与方法14-17    结果17-23    讨论23-25    结论25    参考文献25-29第二部分 前列腺素E_2对急性肝衰竭小鼠肝再生促进作用及其与HGF、Wnt 及β-catenin关系的研究29-53    前言29    材料与方法29-40    结果40-46    讨论46-48    结论48    参考文献48-53综述53-64    参考文献57-64附录64-67硕士期间发表的论文67-68致谢68-70

  本文关键词：前列腺素E_2对急性肝衰竭小鼠肝脏的保护作用及促肝再生作用的研究，，由笔耕文化传播整理发布。