【作者】 周庆兵； 戎光； 李靖； 梅俊； 金宇； 徐凤芹；

【Author】 Zhou Qing-bing;Rong Guang;Li Jing;Mei Jun;Jin Yu;Xu Feng-qing;Institute of Geriatrics, Xiyuan Hospital, Chinese Academy of Chinese Medicine;Shenzhen Second People’s Hospital;

【机构】 中国中医科学院西苑医院老年医学研究所； 深圳市第二人民医院；

【摘要】 目的观察化浊通脉方对ox-LDL诱导的RAW264.7来源泡沫细胞胆固醇逆转运相关基因的表达作用。方法以ox-LDL诱导RAW264.7细胞形成泡沫细胞模型;CCK-8法检测不同浓度化浊通脉方对泡沫细胞的增殖抑制情况;将细胞分为4组,分别为模型组(80μg/mL ox-LDL)、小剂量化浊通脉方组(80μg/mL ox-LDL+化浊通脉方300倍稀释)、中剂量化浊通脉方组(80μg/mL ox-LDL+化浊通脉方120倍稀释)、大剂量化浊通脉方组(80μg/mL ox-LDL+化浊通脉方60倍稀释),处理24h后收集细胞,总胆固醇试剂盒检测化浊通脉方的降脂效应;同时提取RNA,运用荧光定量PCR检测胆固醇代谢相关基因ABCA1、ABCG1、SR-BI以及PPAR-γ的表达变化。结果油红O染色显示,以80μg/mL ox-LDL处理的RAW264.7细胞泡沫化明显;CCK-8法显示化浊通脉方对RAW264.7来源泡沫细胞具有显著的抑制作用;胆固醇检测发现,大剂量化浊通脉能够降低细胞总胆固醇(P=0.045),小剂量及中剂量无降脂作用。(P>0.05)。RT-PCR检测显示,化浊通脉方能够提高ABCA1、SR-BI基因表达(P=0.044;P=0.0028),降低PPAR-γ表达(P=0.0006),对ABCG1基因表达改变不明显(P=0.26)。结论化浊通脉方可能通过提高泡沫细胞中胆固醇逆转运关键基因ABCA1、SR-BI实现抗AS效应。更多还原

【Abstract】 Objective To observe the effects of Huazhuotongmai decoction on the expressions of genes about reverse cholesterol transport( RCT)in RAW264.7 derived foam cells induced by ox-LDL. Methods RAW264.7 cells were induced by ox-LDL to form foam cell model. CCK-8 was used to detect the inhibition effects of huazhuang tongmai recipe on foam cells. Then foam cells were divided into four groups: model group(treated by 80μg/m L ox-LDL only),low-dose group(treated by 80μg/m L ox-LDL and 300 times dilution of huazhuang tongmai recipe), middle-dose group(treated by 80μg/m L ox-LDL and 120 times dilution of huazhuang tongmai recipe), high-dose group(treated by 80μg/m L ox-LDL and 60 times dilution of huazhuang tongmai recipe). After treatement for 24 h, cells were collected. Total cholesterol kits were used to check the change of cholesterol; At the same time, RNA was extracted and the expressions of RCT-related genes ABCA1, ABCG1, SR-BI and PPAR-γwere detected by fluorescence quantitative PCR. Results :Oil red O staining showed that RAW264.7 cells treated with 80 μg/m L ox-LDL turned out to be foam cells well. Huazhuotongmai decoction had a significant inhibitory effect on foam cells through CCK-8 tests.High-dose Huazhuangtongmai could reduce total cholesterol(P=0.045), while low-dose and medium-dose Huazhuangtongmai had no effect on total cholesterol.(P > 0.05). RT-PCR showed that huazhuang tongmai recipe could improve the expressions of ABCA1 and SR-BI genes(P=0.044; P=0.0028) and decreased the expression of PPAR-γ(P=0.0006). There was no significantly change on expression of ABCG1(P=0.26).Conclusion Huazhuotongmai decoction may improve the anti-as effect of key cholesterol reversal genes ABCA1 and SR-BI in foam cells.更多还原