【作者】 郁晨； 刘志华； 潘永明； 徐孝平； 徐剑钦； 杨钦钦； 蔡兆伟； 陈民利；

【Author】 YU Chen;LIU Zhi-hua;PAN Yong-ming;XU Xiao-ping;XU Jian-qin;YANG Qin-qin;CAI Zhao-wei;CHEN Min-li;Laboratory Animal Research Center/Comparative Medical Research Institute,Zhejiang Chinese Medical University;

【机构】 浙江中医药大学动物实验研究中心/比较医学研究所；

【摘要】 目的观察在高脂环境下五指山小型猪、西藏小型猪血脂和炎症细胞的变化,分析冠心病相关血脂和炎症易感基因的表达。方法取雄性五指山小型猪、西藏小型猪各5只,高脂饲喂连续6个月,每月检测血脂和血常规,并检测饲喂前和饲喂6个月时外周白细胞中LDLR、LPL、PCSK9、ApoAV、5-LO、FLAP、MMP-9、ICAM-1、VCAM-1基因和蛋白的表达,及血清中CRP、TNF-α水平;并在饲喂6个月时取腹主动脉组织进行HE染色。结果高脂饲喂后,两种小型猪TC、HDL-C、LDL-C水平均显著升高(P<0.01),西藏小型猪WBC、NEUT%、MONO%和五指山小型猪MONO%亦呈升高趋势(P>0.05);五指山小型猪TC、LDL-C水平高于西藏小型猪,而西藏小型猪的WBC、NEUT%和MONO%显著高于五指山小型猪(P<0.05,P<0.01),且西藏小型猪血清CRP和TNF-α水平显著升高(P<0.05),但五指山小型猪未见显著差异(P>0.05)。高脂饲喂6个月后,两种小型猪均出现明显的AS斑块,五指山小型猪LDLR、ApoAV mRNA和蛋白表达均显著降低(P<0.05,P<0.01),PCSK9、LPL mRNA表达显著升高(P<0.05,P<0.01);西藏小型猪LPL m RNA和蛋白表达均显著升高(P<0.01);五指山小型猪5-LO、FLAP、ICAM-1、VCAM-1 m RNA表达和5-LO蛋白表达均显著升高(P<0.05,P<0.01),西藏小型猪FLAP、MMP-9、ICAM-1、VCAM-1 mRNA和蛋白表达均显著升高(P<0.05,P<0.01),且西藏小型猪MMP-9、ICAM-1、VCAM-1 mRNA和蛋白表达均高于五指山小型猪,而5-LO m RNA和蛋白表达则五指山小型猪高于西藏小型猪。结论高脂环境下五指山小型猪、西藏小型猪均出现不同程度的血脂异常和炎症反应,两种小型猪的冠心病相关血脂和炎症易感基因存在明显异质性,其中,高脂诱导下五指山小型猪外周血白细胞对LDLR、PCSK9、5-LO表达较敏感,而西藏小型猪外周血白细胞中对LPL、ICAM-1、VCAM-1和MMP-9表达较敏感。更多还原

【Abstract】 Objective To observe the changes of blood lipid and inflammatory cells in both WZS minipigs and Tibetan minipigs exposed to high-fat environment,and to analyze the expressions of CAD-related blood lipid and inflammatory susceptibility genes.Methods 5 male WZS minipigs and Tibetan minipigs were selected respectively to feed with high-fat diet for 6 consecutive months.Blood lipids and blood routine examination were detected monthly,and venous blood was taken from before and 6 months after high-fat feeding to test gene and protein expression levels of LDLR,LPL,PCSK9,ApoAV,5-LO,FLAP,MMP-9,ICAM-1 and VCAM-1 in peripheral leukocytes.and the levels of CRP and TNF-α.And the thoracic aorta tissues were taken for HE stained after high-fat feeding for 6 consecutive months.Results After high-fat diet feeding,the levels of TC,HDL-C and LDL-C in two minipigs were all significantly increased(P<0.01),and WBC,NEUT% and MONO%in Tibetan minipigs as well as MONO% in WZS minipigs tended to rise(P>0.05).And TC and LDL-C levels in WZS minipigs were higher than those in Tibetan minipigs,whereas WBC,NEUT%and MONO% in Tibetan minipigs were much higher than those in WZS minipigs(P<0.05,P<0.01).After high-fat feeding for 6 months,there were obvious plaques in the two minipigs,but the degree of AS lesions in Tibetan minipig was more serious,the mRNA and protein expressions of LDLR and ApoAV in WZS minipigs were markedly decreased(P<0.05,P<0.01),whereas mRNA expressions of PCSK9 and LPL were markedly increased(P<0.05,P<0.01).The m RNA and protein expressions of LPL in Tibetan minipigs were obviously increased(P<0.01).Moreover,the mRNA expressions of5-LO,FLAP,ICAM-1 and VCAM-1,and protein expression of 5-LO in WZS minipigs were significantly increased(P<0.05,P<0.01).Meanwhile,mRNA and protein expressions of FLAP,MMP-9,ICAM-1 and VCAM-1 in Tibetan minipigs were markedly increased(P<0.05,P<0.01).Moreover,m RNA and protein expressions of MMP-9,ICAM-1 and VCAM-1 in Tibetan minipigs were all higher than those in WZS minipigs,whereas mRNA and protein expression of 5-LO was on the contrary.Conclusions WZS and Tibetan minipigs all had various degrees of dyslipidemia and inflammatory reaction under high-fat environment.Significant heterogeneity existed in CAD-related blood lipids and inflammatory susceptibility genes.High-fat inducing was susceptible to LDLR,PCSK9 and 5-LO expressions in peripheral leukocytes of WZS minipigs,and sensitive to LPL,ICAM-1,VCAM-1 and MMP-9 expressions in peripheral leukocytes of Tibetan minipigs.更多还原