交泰丸组分配伍改善小鼠胰岛素瘤B细胞脂质沉积的机制研究

唐悦恒 赵莉 邹欣 徐丽君 陆付耳 董慧

摘要 目的：研究交泰丸中主要活性成分小檗碱（BBR）、肉桂酸（CA）及其配伍（BBR/CA）对软脂酸（PA）诱导的NIT-1胰岛B细胞内脂质沉积的影响。方法：将细胞在PA的培养基中培养24 h，建立高脂诱导的细胞内脂肪沉积模型。分为模型组、BBR组、CA组、BBR/CA组、二甲双胍（Met）组，分别给予相应药物干预，另设对照组。油红O染色法评估細胞内脂肪沉积情况;酶法检测细胞内三酰甘油（TG）含量;使用Western Blotting和实时荧光定量PCR（RT-PCR）法检测细胞腺苷酸活化蛋白激酶（AMPK）及其下游脂肪生成和脂肪酸氧化基因的表达水平，包括脂肪酸合成酶（FAS）、乙酰辅酶A羧化酶（ACC）、磷酸化乙酰辅酶A羧化酶（p-ACC）、肉碱酰转移酶-1（CPT-1）、固醇调节元件结合蛋白（SREBP-1c）。结果：PA诱导使NIT-1胰岛B细胞中脂肪沉积明显增加，TG含量显著升高，AMPK蛋白及其下游靶标（如p-ACC、CPT-1等）表达显著降低。同时，AMPK下游脂肪生成基因包括SREBP-1c mRNA、FAS和ACC蛋白表达显著增加。BBR单药和BBR/CA配伍处理优于CA单药，可显著逆转NIT-1胰岛B细胞中上述基因表达水平的变化。结论：在体外，交泰丸活性组分配伍可减少高脂诱导的NIT-1胰岛B细胞内脂肪沉积，其机制可能与减少脂肪合成和增加脂肪氧化有关。

关键词 交泰丸;小檗碱;肉桂酸;NIT-1胰岛B细胞;脂肪

Abstract Objective：To investigate the effects of berberine（BBR） and cinnamic acid（CA），the main active compounds of Jiaotai pills，and the combination of berberine and cinnamic acid（BBR/CA） on palmitic acid（PA）-induced lipid accumulation in NIT-1 pancreatic B cells.Methods：Cells were incubated in culture medium containing PA for 24 hours to establish a model of lipid accumulation induced by high fat.Then treatments with BBR，CA，the combination of BBR and CA（BBR/CA） and Metformin（Met） were performed respectively，and a control group was set up.Intracellular lipid accumulation was assessed by Oil Red O staining and TG content was measured by enzymatic assay.The expression of adenosine monophosphate-activated protein kinase（AMPK） protein and its downstream lipogenic and fatty acid oxidation genes，including fatty acid synthase（FAS），acetyl-CoA carboxylase（ACC），phosphorylation acetyl-CoA carboxylase（p-ACC），carnitine acyl transferase 1（CPT-1） and sterol regulatory element binding protein 1c（SREBP-1c） were determined by Western blot or real time polymerase chain reaction（RT-PCR）.Results：PA induced an obvious lipid accumulation and a significant increase in intracellular TG content in NIT-1 pancreatic B cells.PA also induced a remarkable decrease in AMPK protein expression and its downstream targets such as p-ACC and CPT-1.Meanwhile，AMPK downstream lipogenic genes，including SREBP-1c mRNA，FAS and ACC protein expressions，were increased significantly.Treatments with BBR and BBR/CA，superior to CA，significantly reversed the above genes changes in NIT-1 pancreatic B cells.Conclusion：It can be concluded that in vitro，the active compounds of Jiaotai pills inhibits PA-induced lipid accumulation by decreasing lipogenesis and increasing lipid oxidation in NIT-1 pancreatic B cells.

Keywords Jiaotai pills; Berberine; Cinnamic Acid; NIT-1 pancreatic B cells; Lipid

中图分类号：R587.1文献标识码：Adoi：10.3969/j.issn.1673-7202.2021.19.002

糖尿病（Diabetes Mellitus，DM）是一种由胰岛素分泌缺陷和（或）胰岛素功能缺陷引起的以慢性高血糖为特征的代谢性疾病[1]。根据国际糖尿病联盟（International Diabetes Federation，IDF）報告，2019年全球约有4.63亿成年人患有DM，预计到2045年这一数字将达到7亿[2]。而中国是世界上DM患者数量最多的国家，发病率高达11.2%[3]。……