化瘀祛痰方对ApoE-/-动脉粥样硬化小鼠主动脉巨噬细胞M1极化及糖酵解相关途径的影响OA
Effects of Huayu Qutan Formula on M1 polarization and glycolysis related pathways of aortic macrophages in ApoE-/-atherosclerosis mice
目的:探讨化瘀祛痰方影响ApoE-/-动脉粥样硬化(AS)小鼠主动脉巨噬细胞M1极化及糖酵解相关途径防治AS发生发展的分子机制.方法:将24只ApoE-/-小鼠随机分为3组:模型组、化瘀祛痰方组、瑞舒伐他汀组,给予高脂饲料喂养;8只C57BL/6J小鼠作为正常组,给予普通饲料喂养.饲养8周后,给药组分别给予化瘀祛痰方及瑞舒伐他汀灌胃4周.采用全自动生化分析仪检测小鼠血清血脂水平,HE及油红O染色观察小鼠主动脉组织病理形态学变化,ELISA法检测各组小鼠血清及主动脉白细胞介素(IL)-1β、IL-6、IL-12、乳酸脱氢酶(LDH)、肿瘤坏死因子-α(TNF-α)水平,免疫荧光共定位检测小鼠主动脉LDH、已糖激酶2(HK2)表达情况,RT-qPCR及Western Blot检测各组小鼠主动脉葡萄糖转运蛋白1(GLUT1)、HK2、M2型丙酮酸激酶(PKM2)、磷酸果糖激酶(PFK)、缺氧诱导因子1α(HIF-1α)mRNA及蛋白表达水平,微板法检测血清葡萄糖-6-磷酸脱氢酶(G6PD)、乳酸(LA)水平.结果:与模型组比较,两给药组小鼠血清TC、TG、LDL-C水平显著下降(P＜0.01),HDL-C水平显著上升(P＜0.01);主动脉斑块明显减小,CD86绿色荧光、CD80红色荧光与LDH、HK2粉色荧光发生共定位,且共定位明显降低;血清及主动脉IL-1β、IL-6、IL-12、LDH、TNF-α含量显著下降(P＜0.01);主动脉GLUT1、HK2、PKM2、PFK、HIF-1α mRNA及蛋白表达显著下降(P＜0.01);血清G6PD、LA含量显著下降(P＜0.01).结论:化瘀祛痰方通过纠正血脂代谢异常,干预巨噬细胞糖酵解途径影响巨噬细胞极化,抑制促炎因子生成,发挥抗AS的作用.
Objective:To investigate the molecular mechanism by which the Huayu Qutan Formula inhibits the progression of atherosclerosis(AS)by regulating macrophage M1 polarization and related glycolysis pathways in the aortas of ApoE-/-AS mice.Methods:A total of 24 ApoE-/-mice were randomly divided into three groups:model group,Huayu Qutan Formula group,and rosuvastatin group,they were fed a high-fat diet,while the 8 C57BL/6J mice were fed a standard diet as the normal control group.After 8 weeks of dietary intervention,Huayu Qutan Formula group and rosuvastatin group were administered their respective treatments via gavage for 4 weeks.Serum lipid levels were measured using an automated biochemical analyzer.Pathological changes in the aortic tissues were observed through HE and oil red O staining.Levels of IL-1β,IL-6,IL-12,LDH,and TNF-α in serum and aortic tissues were detected by ELISA.Co-localization of LDH and HK2 in the aorta was examined using immunofluorescence.mRNA and protein expression levels of GLUT1,HK2,PKM2,PFK,and HIF-1α in the aorta were assessed by RT-qPCR and Western Blot.Serum levels of G6PD and LA were measured using a microplate method.Results:Compared with the model group,both the Huayu Qutan Formula group and the rosuvastatin group exhibited significantly reduced serum levels of TC,TG,and LDL-C(P＜0.01)and increased HDL-C(P＜0.01),atherosclerotic plaques were significantly reduced,and co-localization of CD86(green),CD80(red),and LDH(pink),as well as HK2(pink),was markedly decreased,levels of IL-1β,IL-6,IL-12,LDH,and TNF-α in serum and aortic tissues were significantly lowered(P＜0.01),mRNA and protein expression of GLUT1,HK2,PKM2,PFK and HIF-1α in the aorta were significantly downregulated(P＜0.01),serum levels of G6PD and LA were also significantly reduced(P＜0.01).Conclusion:Huayu Qutan Formula exerts anti-atherosclerotic effects by improving dyslipidemia,modulating macrophage glycolysis pathways,influencing macrophage polarization,and inhibiting the production of pro-inflammatory cytokines.
崔运浩;贾连群
辽宁中医药大学,沈阳 110847辽宁中医药大学,沈阳 110847
巨噬细胞糖酵解化瘀祛痰方动脉粥样硬化机制
MacrophageGlycolysisHuayu Qutan FormulaAtherosclerosis(AS)Mechanism
《中华中医药杂志》 2026 (3)
955-960,6
辽宁省科技计划联合计划(应用基础研究项目)(No.2023JH2/101700203),辽宁省科技计划联合计划(技术攻关计划项目)(No.2024JH2/102600194)
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