目的 研究当归与黄芪提取物联合干预对小鼠抗寒能力以及肝脏内质网应激（endoplasmic reticulum stress,ERS）和线粒体自噬蛋白表达的影响。方法 24只8 w龄雄性C57 BL/6小鼠,适应性喂养5 d后,按体重分为C组、M组、Y组,每组8只;C组、M组每日灌胃蒸馏水,Y组灌胃100 mg/kg当归提取物和800 mg/kg黄芪提取物;灌胃14 d后,将M组、Y组小鼠放入4℃下冷暴露24 h,冷暴露结束时测小鼠肛温（核心体温）,收集血清、肝脏,检测血清和肝脏转氨酶水平,进行肝组织病理学观察、ERS和线粒体自噬相关蛋白检测。结果 4℃急性冷暴露显著降低M组小鼠的核心体温（P<0.05）;与M组相比,Y组核心体温显著提高（P<0.05）;M组小鼠肝组织发生轻微水样变性,细胞间隙变大,肝小叶结构紊乱,线粒体肿胀,粗面内质网局部区域模糊,自噬小体减少;血清及肝脏天门冬氨酸氨基转移酶（aspartate aminotransferase,AST）、丙氨酸氨基转移酶（alanine aminotransferase,ALT）活性显著增加（P<0.05）;肝脏中ERS蛋白葡萄糖调节蛋白78（glucose regulated protein78,GRP78）、蛋白激酶R样内质网激酶（PKR-like ER-resident kinase,PERK）、真核翻译起始因子（eukaryotic initiation factor 2 alpha,eIF2α）、p-eIF2α、激活转录因子4（activating transcription factor-4,ATF4）、p-ATF4、C/EBP同源蛋白（C/EBP homologous protein,CHOP）表达显著上调（P<0.05）,线粒体自噬蛋白泛素结合蛋白p62（ubiquitin-binding protein p62,p62）表达显著上调（P<0.05）,含BECN1结构域的自噬相关蛋白1（autophagy-related protein 1 with BECN1 domain,Beclin1）、微管相关蛋白1轻链3（microtubule-associated proteins light chain3,LC3）I与LC3II比值显著下调（P<0.05）;Y组小鼠肝组织结构明显改善,线粒体水肿现象有所改善,可见自噬小体,血清及肝脏AST、ALT活性、肝脏ERS蛋白表达显著下调（P<0.05）,线粒体自噬蛋白p62表达显著下调（P<0.05）,Beclin1、LC3II/I显著上调（P<0.05）。结论 当归与黄芪提取物联合干预可以提高冷暴露小鼠核心体温,可能通过调节肝脏ERS、改善自噬水平缓解肝损伤。
Abstract
Objective To investigate the effects of Angelica sinensis and Astragalus membranaceus extract joint intervention on cold resistance in mice, as well as its influence on endoplasmic reticulum stress (ERS) and mitophagy-related proteins in the liver. Methods Twenty-four 8-week-old male C57BL/6 mice were acclimatized for 5 days and then divided into C group, M group and Y group by body weight, with 8 mice in each group. The C group and M group received distilled water by daily gavage, while the Y group was administered 100 mg/kg Angelica sinensis extract and 800 mg/kg Astragalus membranaceus extract. After 14 days of gavage, mice in M group and Y group were exposed to 4℃ environment for 24 hours. At the end of cold exposure, rectal temperature (core body temperature) was measured. Serum and liver samples were collected. The levels of transaminases in serum and liver were measured. Histopathology observation was conducted, and ERS and mitophagy related proteins were detected in liver. Results 4℃ exposure significantly reduced the core body temperature of mice in the M group (P<0.05). Compared with the M group, core body temperature of the Y group was significantly increased (P<0.05). Liver tissues of M group exhibited mild hydropic degeneration, enlarged intercellular spaces, disordered liver lobule structure, mitochondria swollen, rough endoplasmic reticulum locally blurred, and autophagosomes reduction. Serum and liver alanine aminotransferase (AST) and alanine aminotransferase (ALT) activities were significantly increased (P<0.05). The expressions of ERS-related proteins glucose regulated protein78 (GRP78), PKR-like ER-resident kinase (PERK), eukaryotic initiation factor 2 alpha (eIF2α), p-eIF2α, activating transcription factor-4 (ATF4), p-ATF4 and C/EBP homologous protein (CHOP) in liver tissues was significantly upregulated (P<0.05). The expression of mitophagy protein ubiquitin-binding protein p62 (p62) was significantly increased (P<0.05), while autophagy-related protein 1 with BECN1 domain (Beclin1) and the microtubule-associated proteins light chain3 (LC3）I and LC3II ratio were significantly decreased (P<0.05). In the Y group, liver structure showed significant improvements, mitochondrial edema was alleviated, and autophagosomes were visible. AST and ALT activities in the serum and liver were significantly reduced, and the expression of ERS-related proteins in the liver was significantly downregulated (P<0.05). The expression of mitophagy protein p62 was significantly decreased (P<0.05), while Beclin1 and LC3II/I ratio were significantly increased (P<0.05). Conclusion The Angelica sinensis and Astragalus membranaceus extract joint intervention elevates core body temperature during acute cold exposure, possibly through regulating liver ERS and autophagy to ameliorating liver injury.
关键词
当归 /
黄芪 /
急性冷暴露 /
内质网应激 /
自噬 /
小鼠
Key words
Angelica sinensis /
Astragalus membranaceus /
acute cold exposure /
endoplasmic reticulum stress /
autophagy /
mouse
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