麦冬多糖通过S1P1通路保护OGD条件下细胞生存  

Cell Protection of Polysaccharide from Ophiopogon Japonieus under the Condition of Oxygen Glucose Deprivation through SIPI Signaling Pathway

作　　者：王硕[1] 冯怡[1] 丁侃[2] 王新平[1] 徐德生[3] 

机构地区：[1]上海中医药大学中药现代制剂技术教育部工程研究中心,上海201203 [2]中国科学院上海药物研究所,上海201203 [3]上海中医药大学附属曙光医院,上海200021

出　　处：《药物生物技术》2009年第3期220-224,共5页Pharmaceutical Biotechnology

基　　金：上海市教委重点学科(No.J50302)

摘　　要：为研究1-磷酸鞘氨醇受体1(S1P1)通路在模拟缺血的OGD(氧气和糖剥夺)条件下麦冬多糖MDG-1的细胞保护作用中的角色,以阐明MDG-1细胞保护作用机制。利用RT-PCR和Western blotting实验,研究MDG-1对S1P1通路的调节作用,并利用化学合成siRNA阻断S1P1表达,同时构建S1P1-全长cDNA序列的真核表达载体作为阳性对照,在OGD条件下,用LDH法检测细胞死亡率。结果表明MDG-1具有显著的细胞保护作用,可保护细胞减少OGD诱导的细胞死亡,上调S1P1蛋白表达,而转染siRNA能阻断S1P1表达,并使MDG-1的细胞保护作用受到明显抑制。限制性酶切消化、PCR、测序结果证实S1P1-pcDNA3.1b载体构建成功,转染入S1P1-pcDNA3.1b载体的HEK293细胞S1P1表达增强,并可减少OGD诱导的细胞死亡。这些结果表明MDG-1可能是通过激活S1P1通路,减少OGD诱导的细胞死亡,起到细胞保护作用。To study the cytoprotective effect of a polysaccharide from Ophiopogon japonicus （MDG- 1） and the possible meehanism involved with S1P1 signal pathway. The S1P1 signaling pathways involved in the cytoprotective effect of MDG- 1 was examined by RT-PCR and Western blotting. Blocking the S1P1 gene by chemical synthesized siRNA and constructing the eukafyotic expression vectors containing the coding region of human full length SIP1 gene as a positive control. The cell protective effect and mechanism of MDG- 1 were measured by Oxygen Glucose Deprivation （OGD） induced ischemia. The results showed that MDG - 1 could up-regulate the expression of S1P1 in mRNA and protein level. MDG - 1 could remarkably prevent cells from death induced by OGD and this effect was abolished when cells were transfected with S1P1 siRNA. Meanwhile, the results of the restriction enzyme digestion, PCR and sequencing demonstrated that the vectors were successfully constructed and could also prevent cells from death induced by OGD when cells were transfected with S1P1 - pcDNA3, lb vector. It can be concluded that MDG- 1 probably protects cells from death induced by OGD through S1P1 signal pathway.

关 键 词：麦冬多糖 1-磷酸鞘氨醇受体1 模拟缺血 真核表达载体 SIRNA 

分 类 号：R956[医药卫生—药学]