健脾化痰汤含药血清对胰岛素抵抗3T3-L1脂肪细胞蛋白质表达的影响  

Effect of Serum Containing Jianpi Huatan Decoction(健脾化痰汤) on Protein Expression Profiles of Insulin-Resistant 3T3-L1 Adipocytes

作　　者：李保良[1] 张琪[1] 林冠凯[1] 周燕[1] 吴菁[1] 居凌云[1] 沈璐[1] LI Bao-liang;ZHANG Qi;LIN Guan-kai;ZHOU Yan;WU Jing;JU Ling-yun;SHEN Lu(Changzhou TCM Hospital,Changzhou Jianssu 213003,China)

机构地区：[1]常州市中医医院,江苏常州213003

出　　处：《中医药导报》2018年第16期20-25,共6页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：江苏省中医药科技项目(LZ09114);常州市科技计划项目(CZ20120017)

摘　　要：目的:探究健脾化痰汤含药血清对IR脂肪细胞蛋白质表达谱的影响,阐述其治疗IR相关机理。方法:运用体外试验方法,先将3T3-L1小鼠脂肪前体细胞分化为脂肪细胞,再采用地塞米松法构建IR脂肪细胞模型,再次随机分为IR组(10%空白组血清+1640培养液培养IR脂肪细胞)和健脾化痰汤组(10%健脾化痰汤组血清+1640培养液培养IR脂肪细胞),分别培养48 h,测定培养上清葡萄糖消耗量,同时收集各组细胞,分别提取细胞总蛋白,利用2-DE分离总蛋白,用PDQuest8.0软件分析2-DE图谱,比较组间蛋白表达量,运用MALDI-TOF-MS鉴定差异蛋白点肽谱,通过生物信息学分析确定差异蛋白质及其功能。结果:2组凝胶中有12个显著差异表达蛋白质点,与IR组比较,健脾化痰汤中表达上调有9个,即二氢硫辛酰胺脱氢酶、异戊酰辅酶A脱氢酶、酰基辅酶A结合蛋白、三磷酸腺苷合成酶、酰基辅酶A酰基转移酶2、线粒体H+-ATP合成酶、硫氧还蛋白依赖性过氧化物还原酶、过氧化氢酶、二硫键异构酶6;下调有3个,即硫氧还蛋白、钙网蛋白、α-1-酸性糖蛋白,主要涉及能量代谢和氧化应激等相关蛋白。结论:健脾化痰汤可能通过影响脂肪细胞能量代谢和氧化应激相关蛋白表达,调节其能量代谢和氧化应激功能而改善IR,治疗非酒精性脂肪肝。Objective： To observe protein expression profiles of the insulin-resistant 3T3-IA adipocytes after treating with serum containing Jianpi Huatan Decoction and explore the effect and mechanism of Jianpi Huatan Decoction on the insulin-resistant 3T3-LI adipocytes. Methods： The 3T3-L1 preadipoeytes were cuhured and differentiated to mature adipocytes in vitro. The cell models of insulin-resistance were induced by dexametila- sone. Tile cells in Iil group were cuhured by adding 10% serum of blank group to the culture medium and the （：ells in Chinese medicine group by adding 10% serum of medicine group. After cultured for 48 h, the glucose concentrations in the supernalants of the culture fluid were measured with glucose oxidase method and the total proteins were extraeted from cells. The two-dimensional gel electrophoresis （2-DE） technology was adopted to separate the total protein of cell. The PDQuest software was used to analyze 2-DE images, Matrix-assisted laser desorption/ionization time of flight mass spectrometry, peptide sequence tag and peptide mass fingerprint were applied to identify these protein spots thai have different expression between the two groups. Results： There were 12 dift）rently expressed protein spots （P〈0.05）, among which 9 spots increased, namely, Dihydrolipuamide dehydrogenase, lsovaleryl-CoA dehydrogenase, Acyl-CoA binding protein, ATP synthase, Acetyl-Coenzyme A acetyltransferase 2, Mitochondrial H＋-ATP synthase, Thioredoxin-dependent peroxide reduetase, Catalase and Protein disulfide lsomerase-assoeiated 6, and 3 spots decreasd, including Calretieulin, Alpha-l-acid glycoprotein, Thioredoxin, in Chinese medieine group, compared with IR group. The identified 12 protein spots are the key enzyme related to energy metabolism and oxidative stress. Conclusions： Jianpi Huatan Deeoction could mainly in- fluence the expression of proteins related to energy metabolism and oxidative stress, and regulate the cell energy metabolism and oxidative stress ability to improve the I

关 键 词：胰岛素抵抗 健脾化痰汤 药物血清 蛋白质表达 3T3-L1脂肪细胞 

分 类 号：R285.5[医药卫生—中药学]