银杏苦内酯B对脂多糖诱导的微血管内皮细胞作用的机制  

Mechanism of effect of Ginkgolide B on lipopolysaccharide induced microvascular endothelial cells

作　　者：陈凯 曹卫丽 夏时海 

机构地区：[1]中国武装警察部队后勤学院附属医院肝胆胰脾中心,天津300162

出　　处：《中华胰腺病杂志》2015年第6期389-393,共5页Chinese Journal of Pancreatology

基　　金：国家自然科学基金资助项目（81173393）;天津市应用基础及前沿技术计划重点项目（12JCZDJC25500）;武警后勤学院附属医院基金面上项目（FYM201522）;西青医院院级课题基金面上项目（XQLX201406）

摘　　要：目的观察银杏苦内酯B（BN52021）对胰腺微血管内皮细胞（MSl细胞）的作用，探讨其分子机制。方法采用MTY法确定脂多糖（LPS）抑制MS1细胞存活的最佳浓度和时间，同法确定BN52021增加LPS诱导后MSl细胞存活的最佳浓度。通过实时定量PCR、蛋白质印迹法检测血小板活化因子受体（PAFR）信号通路中腺苷酸环化酶（AC）、磷脂酶A2（PLA2）、磷脂酶c（PLCβ）、酪氨酸蛋白激酶（PTK）和G蛋白耦联受体激酶（GRK）mRNA和蛋白表达量。结果10μg／mlLPS作用24h为抑制MSl细胞存活的最佳浓度和作用时间。50mmol／LBN52021是最佳干预浓度。LPS诱导后MSl细胞株AC、GRK、PLA2、PLCβ、PTKmRNA表达量分别为4．02±0．14、2．63±0．03、3．31±0．12、2．09±0．08、1．85±O．07，较对照组（均为1）明显上调；BN52021干预后MSl细胞AC、GRK、PLA2、PLCβmRNA表达量分别为2．35±0．13、1．17±0．14、1．87±0．11、1．65±0．10，较LPS诱导组显著下调，差异均有统计学意义（P值均〈0．05），PTKmRNA表达量为1．83±0．13，与LPS诱导组差异无统计学意义。蛋白印迹法显示各基因的蛋白表达与mRNA表达的变化一致。结论BN52021能有效地下调LPS诱导MSl细胞后PAFR信号通路中被上调的AC、GRK、PLA2和PLC[3基因表达。Obiective To investigate the effect of Ginkgolide B （BN52021） on lipopolysaccharide （LPS） induced pancreas microvascular endothelialv（ MS1 ） ceils, and to explore its molecular mechanism. Methods The optimal concentration and best time point of LPS inhibing MS1 cell survival and the optimal concentration of BN52021 increasing survival of LPS induced MS1 cells were determined by MTr. The mRNA and protein expression of adenylate cyclase （ AC ） , phospholipase A2 （ PLA2 ）, phospholipase CI3 （ PLCI3 ）, protein tyrosine kinase （PTK） and G protein coupled receptor kinase （GRK） in platelet activating factor receptor（PAFR） signal pathway in MS1 cells were determined by real-time PCR and Western blot. Results It was showed that 10 μg/ml LPS for 24 h was the optimal concentration and best time point to induce the decrease of MS1 cells. 50 mmol/L of BN52021 was the optimal concentration of increaclng survival of LPS induced MS1 ceils. After LPS induction, AC, GRK, PLA2, PLCI3, PTK mRNA expressions of MS1 ceils were 4.02 ±0.14, 2.63 ± 0.03, 3.31 ± 0.12, 2.09 ± 0.08, 1.85 ± O. 07, which were significantly higher than those in control group （P 〈 0.01 ）. After BN52021 treatment, AC, GRK, PLA2, PLC[3 mRNA expressions of LPS induced MS1 cells were 2.35 ±0.13, 1.17 ±0.14, 1.87 ±0.11, 1.65 ±0.10, which were significantly lower than those in LPS induction group （ P 〈 0. 01 ）. The expression of PTK mRNA was 1.83± 0.13, which was not significantly different from that in LPS induction group. Western blot showed that the levels of protein expression were consistent with those of mRNA expression. Conclusions BN52021 can down-regulate the up- regulated genes expression of AC, GRK, PLA2 and PLC[3 in the PAFR signal pathway in LPS induced MS1 cells.

关 键 词：胰腺炎 急性坏死性 银杏苦内酯类 脂多糖类 微血管 内皮细胞 

分 类 号：R285[医药卫生—中药学]