摘要目的:探讨代谢相关脂肪性肝病(MAFLD)大鼠肝脏缺血再灌注损伤(HIRI)与碳水化合物反应元件结合蛋白/缺氧诱导因子-1α(ChREBP/HIF-1α)通路的关系。方法:取40只10周龄SD大鼠，体重220~250 g，分为5组，每组8只，分别为对照组、MAFLD组、HIRI组、HIRI＋ ChREBPα/14-3-3 regulator-1(HIRI＋C1)组和HIRI＋C1＋高糖处理(HIRI＋C1＋Glu)组。高脂高糖饮食喂养建立MAFLD模型，HIRI处理时间为30 min。检测各组大鼠的血清游离脂肪酸(FFA)、甘油三酯、丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)及乳酸脱氢酶(LDH)含量，并计算胰岛素抵抗指数(HOMA-IR)等指标。F4/80和CD86的免疫荧光双重染色用于识别肝脏中的M1型库普弗细胞。蛋白质印迹法检测相关蛋白表达。结果:与对照组相比，MAFLD组大鼠血清FFA、甘油三酯、HOMA-IR升高，肝组织白细胞介素-6(IL-6)、肿瘤坏死因子α(TNF-α)水平以及ChREBP和HIF-1α表达较高，而IL-10水平较低(均 P<0.05)，组织病理学分析提示肝脏脂肪变性。与MAFLD组相比，HIRI组血清FFA、甘油三酯、HOMA-IR、ALT、AST、LDH、肝组织IL-6、TNF-α及IL-10均较高(均 P<0.05)，组织F4/80＋CD86阳性细胞以及ChREBP、HIF-1α和裂解的半胱氨酸蛋白酶-3(Cleaved caspase-3)表达较多(均 P<0.05)，组织学分析显示脂质沉积增多，肝细胞损伤、变性明显。与HIRI组相比，HIRI＋C1组的FFA、甘油三酯、HOMA-IR、ALT、AST及LDH水平显著降低，肝脏组织IL-6、TNF-α和IL-10含量较少，F4/80＋CD86阳性细胞以及ChREBP、HIF-1α和Cleaved caspase-3表达较少(均 P<0.05)，组织学分析显示脂质沉积和肝细胞损伤显著改善。与HIRI＋C1组相比，HIRI＋C1＋Glu组上述指标升高(均 P<0.05)，脂质沉积和肝细胞损伤进一步加重。 结论:ChREBP/HIF-1α信号的激活能够促进MAFLD大鼠HIRI的发生和发展。C1处理可抑制库普弗细胞促炎极化，减轻肝脏组织病理损伤和脂质沉积，其机制可能与抑制ChREBP/HIF-1α通路有关。

abstractsObjective:To investigate the relationship between the ChREBP/HIF-1α pathway and hepatic ischemia-reperfusion injury (HIRI) in rats with metabolic associated fatty liver disease (MAFLD).Methods:Forty 10-week-old Sprague-Dawley rats, weighing 220-250 g, were divided into five groups, with eight rats in each group: control group, MAFLD group, HIRI group, HIRI + ChREBPα/14-3-3 regulator-1 (HIRI+ C1) group, and HIRI+ C1+ high glucose treatment (HIRI+ C1+ Glu) group. The MAFLD model was established by feeding a high-fat, high-sugar diet, and the HIRI treatment duration was 30 min. Serum free fatty acids (FFA), triglycerides, alanine aminotransferase (ALT), aspartate amino-transferase (AST), and lactate dehydrogenase (LDH) levels were measured, and the homeostasis model assessment of insulin resistance (HOMA-IR) was calculated. Immunofluorescence dual staining for F4/80 and CD86 was used to identify M1-type Kupffer cells in the liver. Western blotting was used to detect the expression of related proteins.Results:Compared to the control group, the MAFLD group exhibited significantly elevated serum levels of FFA and triglycerides, as well as increased HOMA-IR (all P<0.05). Furthermore, the MAFLD group showed higher hepatic levels of IL-6 and TNF-α, along with increased expression of ChREBP and HIF-1α, while IL-10 levels were significantly lower (all P<0.05). Histopathological analysis confirmed the presence of hepatic steatosis in the MAFLD group. Compared to the MAFLD group, the HIRI group displayed significantly higher serum levels of FFA, triglycerides, HOMA-IR, ALT, AST, and LDH, as well as increased hepatic levels of IL-6, TNF-α, and IL-10 (all P<0.05). The HIRI group also showed increased numbers of F4/80+ CD86 positive cells in liver tissue, along with elevated expression of ChREBP, HIF-1α, and cleaved caspase-3 (all P<0.05). Histological analysis of the HIRI group revealed exacerbated lipid deposition, accompanied by pronounced hepatocyte injury and degeneration. Notably, the HIRI+ C1 group, when compared to the HIRI group, demonstrated a significant reduction in serum FFA, triglycerides, HOMA-IR, ALT, AST, and LDH levels, along with decreased hepatic levels of IL-6, TNF-α, and IL-10. Furthermore, levels of F4/80+ CD86 positive cells, ChREBP, HIF-1α, and cleaved caspase-3 were also significantly reduced in the HIRI+ C1 group (all P<0.05), and histological analysis revealed a marked improvement in both lipid deposition and hepatocyte injury. Compared to the HIRI+ C1 group, the HIRI+ C1+ Glu group showed increased levels of the above indicators (all P<0.05), with further exacerbation of lipid deposition and hepatocyte injury. Conclusion:Activation of the ChREBP/HIF-1α signaling pathway can promote the occurrence and progression of HIRI in MAFLD rats. C1 treatment can inhibit pro-inflammatory polarization of Kupffer cells, reduce liver tissue pathological damage and lipid deposition, potentially through inhibiting the ChREBP/HIF-1α pathway.