摘要
目的:考察小檗碱降血胆固醇作用及对肝脏胆固醇7α-羟化酶(CYP7A1)和小肠尼曼-匹克C1型类似蛋白1(NPC1L1)相关基因及蛋白表达的影响。方法:采用高脂饮食诱导建立高脂血症SD大鼠模型,不同剂量的小檗碱给药治疗4 w;检测大鼠肝脏、粪便及血清中总胆固醇(TC)水平及血清低密度脂蛋白胆固醇(LDL-C)水平以评价小檗碱的降脂效果;采用TBA比色试剂盒测定肝脏匀浆和粪便提取物中总胆汁酸(TBA)浓度;采用HPLC法检测大鼠肝脏微粒体CYP7A1酶活性;实时荧光定量PCR(q-PCR)检测法及蛋白印迹法(Western blot)检测CYP7A1、NPC1L1及与其调节通路相关联基因的表达水平。结果:小檗碱能显著降低高脂血症大鼠肝脏和血清TC水平,降低血清LDL-C,升高粪便中TC水平,并显著增加肝脏及粪便中TBA水平,同时显著上调大鼠肝脏中PPARα、HNF1α、SREBP2、LXRα、CYP7A1的表达和下调小肠中SREBP2、NPC1L1、HNF1α的表达。结论:小檗碱有降胆固醇作用,其机制可能与通过调节LXRα和SREBP2等核因子,进而上调肝脏CYP7A1和下调小肠和NPC1L1的表达有关。
Objective:To investigate the cholesterol removal effect of berberine and its effect on regulating the expressions of liver cholesterol 7α-hydroxylase(CYP7 A1),small intestine Nieman-Pick C1-like protein 1(NPC1 L1)and their related genes and proteins.Methods:SD rats with hyperlipidemia were induced by high-fat diet and administered with different doses of berberine for 4 weeks;The levels of total cholesterol(TC)in liver,feces and serum and low density lipoprotein cholesterol(LDL-C)in serum of rats were measured to evaluate the lipid lowering effect of berberine.TBA colorimetric kit was used to determine the total bile acid(TBA)concentration in liver homogenates and fecal extracts.The activity of CYP7 A1 enzyme in rat liver microsomes was detected by HPLC.The expression levels of CYP7 A1 and NPC1 L1 as well as the genes and proteins associated with their regulatory pathways were detected by real-time quantitative PCR(q-PCR)and Western blot.Results:Berberine significantly reduced TC levels in the liver and serum of hyperlipidemia rats,decreased the level of LDL-C in the serum,increased TC levels in the stool,and significantly increased TBA level in the liver and feces.Meanwhile,the expressions of PPARα,HNF1α,SREBP2,LXRαand CYP7 A1 in the liver of rats were significantly up-regulated,and the expressions of SREBP2,NPC1 L1,HNF1αin the small intestine were down-regulated.Conclusion:Berberine has the cholesterol removal effect,and the mechanism may be related to the up-regulation of liver CYP7 A1 and down-regulation of small intestine NPC1 L1 by regulating nuclear factors such as LXRαand SREBP2.
作者
张东行
郭彩娟
方沐潮
贝伟剑
郭姣
ZHANG Dong-xing;GUO Cai-juan;FANG Mu-chao;BEI Wei-jian;GUO Jiao(Institute of Traditional Chinese Medicine,Guangdong Pharmaceutical University/Guangdong Metabolic Disease Research Center of Integrated Chinese and Western Medicine,Guangzhou 510006,China)
出处
《中药材》
CAS
北大核心
2021年第2期416-421,共6页
Journal of Chinese Medicinal Materials
基金
国家自然科学基金重点资助项目(81173626)
广东省联合培养研究生示范基地项目
广药大“药创未来”众创空间建设(20161315)
2017年中央支持地方高校发展专项资金项目(201708)
