蚯蚓活性组分对四氯化碳诱导小鼠内质网应激所致急性肝损伤的保护作用(1)
[摘要]該研究旨在探讨蚯蚓活性组分(EWAs)对四氯化碳(CCl4)诱导小鼠内质网应激(ERS)所致急性肝损伤的保护作用。腹腔注射10%CCl4制备内质网应激所致急性肝损伤模型；血清生化指标检测ALT，AST，SOD，GSHPX酶活性及MDA含量变化；计算肝、脾指数；HE染色观察肝脏病理学变化；TUNEL法检测肝组织细胞凋亡情况；免疫组织化学法检测ERS标志性蛋白GRP78和CHOP表达情况；Western blot法检测ERS相关蛋白的表达水平。结果显示，与模型组小鼠相比，EWAs的高、中、低剂量组血清学各项指标均有显著改善(P<005或P<001)，肝脏病变范围减小，且损伤程度明显减轻，小鼠肝脏指数和脾脏指数均有明显变化(P<005或P<001)；各剂量给药组的肝组织细胞凋亡指数明显下降(P<005或P<001)；肝组织中ERS相关蛋白GRP78和CHOP表达水平显著降低(P<005或P<001)，各剂量给药组均能显著下调GRP78和CHOP以及CHOP上游信号通路PERKeIF2αATF4的蛋白表达(P<005或P<001)。综上，EWAs对ERS所致的小鼠急性肝损伤具有显著保护作用，其机制可能通过抑制氧化应激和ERS，从而减轻肝脏损伤，并可下调ERS标志性凋亡蛋白CHOP表达，抑制细胞凋亡实现的。
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[关键词]蚯蚓活性组分； 内质网应激； 肝损伤； GRP78； CHOP
[Abstract]To study the protective effect of earthworm active ingredients(EWAs) against endoplasmic reticulum stress(ERS)induced acute liver injury in mice. The model of liver injury was induced through intraperitoneal injection of 10%CCl4. Serum glutamicpyruvic transaminase(ALT)， glutamicoxaloacetic transaminase(AST)， superoxide dismutase(SOD) and glutathione peroxidase(GSHPX) activity and malondialdehyde(MDA) concentration were detected by colorimetric method. Histological examination was performed through hematoxylineosin staining and light microscopy， and apoptosis was detected using terminal transferase dUTP nick end labeling. The expressions of ERS related proteins， including glucose regulated protein 78(GRP78)， protein kinase Rlike ER kinase(PERK)， eukaryotic transcription initiation factor 2α(eIF2α)， active transcription factor4(ATF4) and CCAAT/enhancer binding homologous protein(CHOP)， were measured by immunohistochemistry and Western blot. According to the results， compared with the model group，serological indexes in the high， middle and low doses of EWAs were significantly improved (P<0.05 or P<0.01)， the extent of liver lesion was decreased and the degree of injury was significantly reduced， and that the liver index and the spleen index of mice were significantly changed(P<0.05 or P<0.01). In liver tissue， the expressions of GRP78 and CHOP were significantly decreased(P<0.05 or P<0.01). The protein expressions of GRP78， CHOP and its upstream signaling pathway PERKeIF2ATF4 were significantly decreased in each dose group(P<0.05 or P<0.01). In summary， EWAs has a significant protective effect on ERSinduced acute liver injury， and its mechanism may be correlated with the inhibition of oxidative stress and ERS， and downregulation of ERS marker protein CHOP expression， andinhibition of apoptosis.
[Key words]EWAs； ERS； liver injury； GRP78； CHOP, http://www.100md.com(赵亚飞 高杨 吴欣芳 王建刚 段冷昕)
, http://www.100md.com
[关键词]蚯蚓活性组分； 内质网应激； 肝损伤； GRP78； CHOP
[Abstract]To study the protective effect of earthworm active ingredients(EWAs) against endoplasmic reticulum stress(ERS)induced acute liver injury in mice. The model of liver injury was induced through intraperitoneal injection of 10%CCl4. Serum glutamicpyruvic transaminase(ALT)， glutamicoxaloacetic transaminase(AST)， superoxide dismutase(SOD) and glutathione peroxidase(GSHPX) activity and malondialdehyde(MDA) concentration were detected by colorimetric method. Histological examination was performed through hematoxylineosin staining and light microscopy， and apoptosis was detected using terminal transferase dUTP nick end labeling. The expressions of ERS related proteins， including glucose regulated protein 78(GRP78)， protein kinase Rlike ER kinase(PERK)， eukaryotic transcription initiation factor 2α(eIF2α)， active transcription factor4(ATF4) and CCAAT/enhancer binding homologous protein(CHOP)， were measured by immunohistochemistry and Western blot. According to the results， compared with the model group，serological indexes in the high， middle and low doses of EWAs were significantly improved (P<0.05 or P<0.01)， the extent of liver lesion was decreased and the degree of injury was significantly reduced， and that the liver index and the spleen index of mice were significantly changed(P<0.05 or P<0.01). In liver tissue， the expressions of GRP78 and CHOP were significantly decreased(P<0.05 or P<0.01). The protein expressions of GRP78， CHOP and its upstream signaling pathway PERKeIF2ATF4 were significantly decreased in each dose group(P<0.05 or P<0.01). In summary， EWAs has a significant protective effect on ERSinduced acute liver injury， and its mechanism may be correlated with the inhibition of oxidative stress and ERS， and downregulation of ERS marker protein CHOP expression， andinhibition of apoptosis.
[Key words]EWAs； ERS； liver injury； GRP78； CHOP, http://www.100md.com(赵亚飞 高杨 吴欣芳 王建刚 段冷昕)