壳寡糖上调SRBI核心岩藻糖基化修饰水平减弱小鼠动脉粥样硬化斑块形成  

Chitosan oligosaccharides alleviates the formation of atherosclerotic plaque through up-regulating core-fucosylation of SRBI in mice

作　　者：陈林木 黄云秀 CHEN Linmu;HUANG Yunxiu(Department of Clinical Pharmacy,Zhongshan People’s Hospital,Zhongshan 528403,China;Department of Laboratory Medicine,Zhongshan People’s Hospital,Zhongshan 528403,China)

机构地区：[1]中山市人民医院药学部,广东中山528403 [2]中山市人民医院检验医学中心,广东中山528403

出　　处：《基础医学与临床》2024年第4期474-482,共9页Basic and Clinical Medicine

基　　金：广东省医学科研基金(B2020116);中山市第三批社会公益与基础研究专项(2020B3004);中山市人民医院一类青年骨干科研启动经费(SGKY2022006)。

摘　　要：目的 从蛋白糖基化修饰角度探讨壳寡糖(COS)减弱动脉粥样硬化(AS)斑块形成的机制。方法 将40只ApoE-/-小鼠随机分为对照组和COS组。对照组饲给予高脂饲料12周,COS组给予高脂饲料+COS(每天灌胃,500 mg/kg)12周。全自动生化分析仪检测两组小鼠的血脂水平并对主动脉斑块切片进行HE和油红O染色。凝集素芯片、液相色谱-串联质谱、酶联免疫吸附试验检测血清中糖蛋白变化。胆固醇酯流出实验和游离胆固醇酯测定实验评价清道夫受体B类成员1(SRBI)糖基化修饰位点改变对巨噬细胞胆固醇流出量的影响。结果 COS显著降低小鼠TC和LDL-C水平(P<0.05),但对TG、HDL-C、ApoA1和ApoB100无影响。与对照组相比,COS组主动脉内膜厚度和斑块大小明显变薄、变小(P<0.05)。两组小鼠血清中变化最明显的小扁豆凝集素(LCA)结合蛋白分子量为80~90 ku,清道夫受体B类成员1为其中之一。COS促进了细胞内胆固醇的流出,抑制了游离胆固醇酯的积累(P<0.05)。SRBI敲低或N108糖基化位点突变可抑制COS引起的胆固醇流出,增加细胞内游离胆固醇的积累(P<0.05)。结论 COS通过增加SRBI糖基化水平促进脂质外排,从而减少动脉粥样硬化的形成。Objective To explore the mechanism of chitosan oligosaccharides(COS)in reducing atherosclerotic plaque formation from the perspective of protein glycosylation modification.Methods Totally 40 ApoE-/-mice were randomly divided into control group and COS group.The control group was given a high-fat diet for 12 weeks,and COS group was given a high-fat diet plus COS(gavage per day,500 mg/kg)for 12 weeks.Serum lipid detection,HE staining and Oil red O staining were used to detect plaque formation.Lectin chip,liquid chromatography tandem-mass spectrometry and ELISA were used to detect potential changes of glycoprotein in serum.Cholesterol ester outflow and free cholesterol ester determination experiment were used to evaluate the effect of changes in scavenger receptor class B typeⅠ(SRBI)protein glycosylation modification site on cholesterol effluence in macrophages.Results COS significantly reduced the level of TC and LDL-C(P<0.05)in mice,but had no effect on the level of TG,HDL-C,ApoA1 and ApoB100.The intima thickness and plaque size of the aorta were significantly thinner and smaller(P<0.05)in the COS group compared with the control group.The molecular weight of lens culinaris agglutinin(LCA)binding protein with the most obvious change is 80-90 ku,and SRBI was one of them.COS promoted the cholesterol outflow and inhibited the accumulation of free cholesterol ester in RAW264.7 cell(P<0.05).Knockdown or glycosylation site mutation with SRBI inhibited cholesterol outflow caused by COS,and increased the accumulation of intracellular free cholesterol(P<0.05).Conclusions COS promotes lipid efflux by increasing SRBI glycosylation and expression,thereby alleviating atherosclerotic plaque formation.

关 键 词：壳寡糖 清道夫受体B类成员1 核心岩藻糖基化 动脉粥样硬化 

分 类 号：R543.5[医药卫生—心血管疾病]