千金子制霜前后提取物通过肝X受体-腺苷三磷酸结合盒转运体A1信号通路对Caco-2细胞中胆固醇外流的影响  

Influence of extracts from Euphorbiae Semen before and after frosting on cholesterol efflux in Caco-2 cells through liver X receptor-adenosine triphosphate-binding cassette transporter A1 pathway

作　　者：王佩华 岳珠珠 魏晓彤 王梦琳 石双慧 王英姿[1] WANG Pei-hua;YUE Zhu-zhu;WEI Xiao-tong;WANG Meng-lin;SHI Shuang-hui;WANG Ying-zi(College of Chinese Meteria Medica,Beijing University of Chinese Medicine,Beijing 102488,China)

机构地区：[1]北京中医药大学中药学院,北京102488

出　　处：《中国临床药理学杂志》2023年第2期201-205,共5页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(82074021)。

摘　　要：目的 研究千金子制霜前后提取物通过肝X受体(LXR)-腺苷三磷酸结合盒转运体A1(ABCA1)信号通路对Caco-2细胞中胆固醇外流的影响差异。方法 将Caco-2细胞分为7组:对照组、千金子生品提取物低、中、高剂量组和千金子霜品提取物低、中、高剂量组。对照组仅加入细胞培养液,低、中、高剂量组分别加入生药量为50,200和800μg·mL^(-1)的千金子生品或霜品提取物。用实时荧光定量聚合酶链反应检测LXRα和ABCA1 mRNA的表达水平,用胆固醇流出实验检测胆固醇外流率。结果 千金子生品提取物低、高剂量组和千金子霜品提取物低、高剂量组及对照组的LXRαmRNA相对表达水平分别为0.37±0.03,0.13±0.01,0.53±0.05,0.48±0.03和1.00±0.05,ABCA1 mRNA相对表达水平分别为0.25±0.02,0.06±0.01,0.63±0.06,0.32±0.02和1.01±0.15,胆固醇流出率分别为(37.11±0.21)%,(29.98±0.44)%,(48.38±0.76)%,(37.66±0.25)%和(48.69±0.82)%。千金子生品提取物低、高剂量组的上述指标与对照组相比,差异均有统计学意义(均P<0.001);在相同剂量浓度下,千金子霜品提取物组的上述指标与千金子生品提取物组相比,差异均有统计学意义(P<0.05,P<0.001)。结论 千金子生品提取物可能通过抑制LXR-ABCA1信号通路影响膜脂成分胆固醇的流出,而制霜后作用明显减弱,提示千金子制霜减毒作用机制可能与LXR-ABCA1信号通路调控胆固醇外流有关。Objective To study the functional difference of extracts from Euphorbiae Semen before and after frosting cholesterol efflux in Caco-2 cells through liver X receptor(LXR)-adenosine triphosphate-binding cassette transporter A1(ABCA1) pathway. Methods Caco-2 cells were divided into 7 groups: control group, Euphorbiae Semen-L,-M,-H groups and Euphorbiae Semen Pulveratum-L,-M,-H groups. The control group was only added with cell culture medium, and low, middle, high dose groups were added with crude drug doses of 50, 200 and 800 μg·mL^(-1)of Euphorbiae Semen extract or Euphorbiae Semen Pulveratum extract. Real-time polymerase chain reaction was applied to detect the mRNA expressions of LXRα and ABCA1 in Caco-2 cells. Cholesterol efflux assay was used to detect the cholesterol efflux rate in Caco-2 cells. Results The relative expressions of LXRα mRNA in Euphorbiae Semen-L,-H groups,Euphorbiae Semen Pulveratum-L,-H groups and control group were 0. 37 ± 0. 03,0. 13 ± 0. 01,0. 53 ± 0. 05,0. 48 ± 0. 03 and 1. 00 ± 0. 05;the relative expressions of ABCA1 mRNA were 0. 25 ± 0. 02,0. 06 ± 0. 01,0. 63 ± 0. 06,0. 32 ± 0. 02 and 1. 01 ± 0. 15;the cholesterol efflux rates were( 37. 11 ± 0. 21) %,( 29. 98 ± 0. 44) %,( 48. 38 ± 0. 76) %,( 37. 66 ± 0. 25) % and( 48. 69 ± 0. 82) %,respectively. There were significant differences in above indexes between the Euphorbiae Semen-L,-H groups and the control group( all P < 0. 001). At the same dose concentration,there were significant differences in above indexes between the Euphorbiae Semen Pulveratum groups and Euphorbiae Semen groups( P < 0. 05,P < 0. 01). Conclusion The extract of Euphorbiae Semen could affect the cholesterol efflux of membrane lipid component by inhibiting the LXR-ABCA1 pathway,while the effects were obviously weakened after the frosting. It suggested that the attenuating toxicity mechanism of frosting for Euphorbiae Semen may be related to the regulation of cholesterol efflux by LXR-ABCA1 signaling pathway.

关 键 词：千金子 肝X受体 腺苷三磷酸结合盒转运体A1 胆固醇外流 CACO-2细胞 制霜减毒 

分 类 号：R28[医药卫生—中药学]