番石榴叶总三萜改善3T3-L1脂肪细胞胰岛素抵抗  

Total triterpenoids from Psidium Guajava leaf improves insulin resistance in 3T3-L1 adipocytes

作　　者：李秀存[1] 马锦锦[1] 赵晶晶[2] 叶开和[1] 吕艳青[1] 王小康[1] 魏崧丞 张晓琦[3] 叶春玲[1] 

机构地区：[1]暨南大学药学院药理学教研室,广东广州510632 [2]广东省妇幼保健院药学部,广东广州511400 [3]暨南大学中药与天然药物研究所,广东广州510632

出　　处：《中国病理生理杂志》2016年第2期314-320,共7页Chinese Journal of Pathophysiology

基　　金：广东省高等学校科技创新重点项目(No.cxzd1111)

摘　　要：目的：观察番石榴叶总三萜（TTPGL）对3T3-L1脂肪细胞胰岛素抵抗（IR）的改善作用,并探讨其可能的作用机制。方法：培养3T3-L1前脂肪细胞并诱导其分化,给予TTPGL（0.3、1、3、10μg/L）,并设溶媒（0.1%DMSO）组、阳性药正钒酸钠（Van,10μmol/L）组、正常对照（control）组和模型（model）组,药物作用48 h。MTT法检测药物对前脂肪细胞活力的影响,油红O染色法观察其对细胞分化的影响。建立IR模型后,药物处理48 h,葡萄糖氧化酶-过氧化物酶法（GOD-POD）检测IR脂肪细胞上清液中葡萄糖消耗量;比色法检测游离脂肪酸（FFA）水平;ELISA法检测脂肪因子分泌水平;real-time PCR检测IR脂肪细胞蛋白酪氨酸激酶1B（PTP1B）的mRNA表达量;Western blot检测磷酸化胰岛素受体底物1/胰岛素受体底物1（p-IRS-1/IRS-1）和磷酸化蛋白激酶B/蛋白激酶B（p-Akt/Akt）的蛋白水平。结果：与溶媒组比较,TTPGL显著提高了前脂肪细胞的活力并抑制其分化（P〈0.01）。与IR溶媒组比较,无论在基础状态下还是胰岛素刺激状态下,TTPGL（1-10μg/L）均显著地促进了IR脂肪细胞葡萄糖消耗（P〈0.01）;TTPGL（0.3～3μg/L）显著抑制FFA的产生（P〈0.01）。与模型组比较,TTPGL（0.3和3μg/L）显著增加IR脂肪细胞脂联素的分泌（P〈0.05）并抑制TNF-α的分泌（P〈0.01）,TTPGL（3μg/L）对抵抗素的分泌有显著抑制作用（P〈0.05）,对瘦素分泌无显著作用;TTPGL（3μg/L）显著下调IR脂肪细胞PTP1B的mRNA表达（P〈0.01）;TTPGL（3μg/L）极显著上调p-IRS-1/IRS-1的水平;TTPGL（0.3和3μg/L）显著上调p-Akt/Akt的蛋白水平（P〈0.05）。结论：TTPGL具有显著改善3T3-L1脂肪细胞IR的作用,其作用机制可能与TTPGL下调了IR脂肪细胞PTP1B mRNA的表达、同时上调p-IRS-1/IRS-1和p-Akt/Akt的蛋白水平有关。AIM： To investigate the effects of total triterpenoids from Psidium guajava leaf （TFPGL） on 3T3- L1 adipocyte insulin resistance （1R） and to explore the possible mechanism. METHODS： 3T3-L1 pre-adipocytes were cultured and induced to differentiate into 3T3-L1 adipocytes, then treated with TrPGL （0.3, 1, 3, 10 μg/L） for 48 h. The cells were divided into 0. 1% DMSO group, positive drug sodium orthovanadate （Van, 10 μmol/L） group, model group and control group. The effect of 3TPGL on the cell activity of pre-adipocytes was detected by MTT assay and its influ- ence on the cellular differentiation was observed by oil red O staining. The IR model of the 3T3-L1 adipocytes was estab- lished successfully and then treated with different drugs for 48 h. The glucose consumption in the supernatant of IR adipo- cyte＇s cuhm＇e medium was assayed by glucose oxidase-peroxidase （GOD-POD）, free fatty acid （FFA） levels were meas- ured by colorimetric method, and adipocytokines levels were assayed by ELISA. The mRNA expression of protein tyrosine phosphatase-1 B （PTP1 B） of IR adipocyte was detected by real-time PCR. The protein levels of phosphorylated insulin re- ceptor substrate 1/insulin receptor substrate 1 （p-IRS-1/IRS-1） and phosphorvlated protein kinasc B/protein kinase B （ p-Akt/Akt） were determined by Western blot. RESULTS： Compared with DMSO group, TYPGL treatment significantly pro- moted the cell activity of 3T3-L1 pre-adipocytes and inhibited its differentiation （ P 〈 0.01 ）. TrPGL （ 1 - 10 μg/L） im- proved glucose consumption of IR adipocytes significantly （ P 〈 0.01 ）, with or without insulin stimulation, and TI＇PGL （0. 3 ~ 3 μg/L） restrained FFA production remarkably（P 〈0. 01 ）. Compared with model group, TI＇PGL （0. 3 and 3 μg/ L） significantly increased the secretion of adiponectin in IR adipocytes （P 〈 0. 05 ）, and inhibited the secretion of tumor necrosis factor-α （TNF-α） （ P 〈0. 01 ）. TI＇PGL （ 3 μg/L） restra

关 键 词：番石榴叶总三萜 胰岛素抵抗 蛋白酪氨激酶1B 胰岛素受体底物1 蛋白激酶B 

分 类 号：R285[医药卫生—中药学] R363[医药卫生—中医学]