家蚕30K蛋白对人肝细胞脂滴蓄积的影响  

Effect of Silkworm(Bombyx mori) 30K Apolipoprotein on Human(Homo sapiens) Hepatic Lipid Accumulation

作　　者：汪凤梅[1] 林樟芽 陈生稳 陈滨[2] 于威[2] 

机构地区：[1]浙江大学医学院附属妇产科医院,杭州310006 [2]浙江理工大学生命科学学院,杭州310018

出　　处：《农业生物技术学报》2015年第12期1542-1549,共8页Journal of Agricultural Biotechnology

基　　金：国家高技术研究发展计划(863)项目(No.2011AA100603);浙江省自然科学基金项目(No.Y3090339)

摘　　要：家蚕(Bombyx mori)30K蛋白是家蚕五龄幼虫血淋巴中含量最高的一类蛋白质,具有多种生物学功能。30K蛋白作为一类昆虫载脂蛋白,推测其可能在哺乳动物细胞中也具有相似的功能,但迄今尚未见报道。为了研究其对哺乳动物细胞内脂类蓄积的影响,本研究利用含诱导液油酸(oleic acid,OA)和棕榈酸(palmic acid,PA)的培养基(OA∶PA=2∶1)诱导正常人(Homo sapiens)肝细胞系L-02 24 h,在体外建立了人脂肪肝细胞模型,并利用该模型分析了两种家蚕30K蛋白30Kc6和Lip-19G1对肝细胞内脂滴蓄积的影响。结果表明,当采用油红O染色法进行检测时,经30Kc6蛋白预处理后的正常肝细胞再经诱导液诱导后,细胞内脂滴的积分光密度(integral optical density,IOD)值/细胞总面积值与未经蛋白处理的对照组细胞相比明显减少(P<0.05),而Lip-19G1蛋白预处理组差异不显著;此外,经30Kc6或Lip-19G1蛋白处理后的脂肪肝细胞模型内脂滴IOD值/细胞总面积值与未经蛋白处理的对照组细胞相比也明显减少(P<0.01)。上述结果表明,家蚕30Kc6蛋白对肝细胞内脂滴蓄积具有一定的预防作用,同时家蚕30Kc6和Lip-19G1蛋白均可以有效减少脂肪肝细胞模型内脂滴的蓄积量。本研究将为家蚕30K蛋白载脂功能的深入研究提供基础信息,并为其作为一种新型的降脂药物开发提供理论依据。Silkworm（Bombyx mori） 30 K proteins are the major protein in the fifth instar larval hemolymph and possess multiple biological functions, such as inhibiting cell apoptosis induced by peroxide, virus or chemically and apolipoprotein functions. As a kind of insect apolipoprotein, they can bind with lipids and transport lipids inside insect larval hemolymph. Therefore it＇s very likely that silkworm 30 K protein also has a similar function in mammalian cells, but it has not been reported so far. In order to study the effect of silkworm 30 K apolipoprotein on lipid metabolism in mammalian cells, oleic acid（OA） and palmitic acid（PA）（OA∶PA=2∶1） solution was used to induce the normal human（Homo sapiens） liver cell line L-02 for 24 h to establish the fatty liver cell model in vitro. Then, the model was used to analyze the effects of these two kinds of silkworm 30 K proteins, 30Kc6 and Lip- 19G1, on lipid accumulation in liver cells. In order to understand the prevention function of these two proteins on lipid accumulation in liver cells, the normal liver cells were pretreated with 30Kc6 or Lip- 19G1 protein before induction and then the cells were used to detect the lipid droplets IOD（integral optical density） value intracellular/total cell area by oil red O staining. The results showed that there was significant difference between 30Kc6 pretreatment group with control group（P〈0.05）.However, there was no difference between Lip- 19G1 pretreatment group with control groups. Moreover, in order to understand the treatment effects of these two proteins on lipid accumulation in the fatty liver cells, the fatty liver cells were treated with 30Kc6 or Lip-19G1 protein, respectively. The fatty liver cells with 30Kc6 or Lip-19G1 protein treatment, the lipid droplets IOD value intracellular/ total cell area all reduced significantly compared with the cells without treatments（P〈0.01）. These results showed that 30Kc6 had a certain inhibitory effects on the accumulation of lipid in human hepatic

关 键 词：家蚕 30K载脂蛋白 脂肪肝细胞模型 油红O染色 

分 类 号：Q581[生物学—生物化学]