摘要
目的探讨Sonic Hedgehog(Shh)-聚腺苷二磷酸核糖聚合酶1[poly(ADP-ribose)polymerase 1,PARP-1]信号通路在茶多酚拮抗胰岛微血管内皮细胞脂毒性中的调控作用。方法以小鼠胰岛微血管内皮MS-1细胞为研究对象,分为正常对照组、溶剂对照组、脂肪酸(0.25mmol/L软脂酸+0.5mmol/L油酸)组、茶多酚(25μmol/L)组、脂肪酸+茶多酚组、PARP-1抑制剂(8μmol/L BYK204165)+脂肪酸组、PARP-1抑制剂+脂肪酸+茶多酚组、Shh抑制剂(2.5μmol/L环巴胺)+脂肪酸组、Shh抑制剂+脂肪酸+茶多酚组及Shh抑制剂+PARP-1抑制剂+脂肪酸+茶多酚组,分别检测各组细胞活力、凋亡水平、一氧化氮(NO)合成及氧化应激相关指标的改变。结果脂肪酸处理后,MS-1细胞存活率下降,细胞凋亡率增高(P<0.05);同时,细胞内NO的含量及总一氧化氮合酶(tNOS)、诱导型NOS(iNOS)和结构型NOS(cNOS)的活性均升高(P<0.05);而且,脂质过氧化产物丙二醛(MDA)含量增加(P<0.05),抗氧化物质谷胱甘肽(GSH)和超氧化物歧化酶(SOD)的水平下降(P<0.05),并增强了PARP-1和磷酸化Shh的表达水平(P<0.05)。茶多酚干预后,各项指标的水平均得以改善(P<0.05);而且,利用BYK204165和环巴胺预处理1h后,茶多酚对脂肪酸的拮抗效应更为显著,各项检测指标与正常对照组比较差异无统计学意义(P>0.05)。结论脂肪酸可诱发胰岛微血管内皮功能损伤,茶多酚具有拮抗脂肪酸毒性的作用,且抑制Shh-PARP-1信号通路能增强茶多酚的保护效应。
Objective To investigate the role of Shh-PARP-1 signaling pathway in the protective effects of tea polyphenols against the fatty acid-induced islet microvessel endothelial function injury. Methods Mouse islet microvessel endothelial MS-1 cells were used in this study, and the cells were divided into normal control group, solvent group, fatty acid group （0.25 mmol/L palmitic acid＋0.5 mmol/L oleic acid）, tea polyphenols group （25 μmol/L）, fatty acid＋tea polyphenols group, PARP-1 inhibitor （8 μmol/L BYK204165）＋fatty acid group, PARP-1 inhibitor＋fatty acid＋tea polyphenols group, Shh inhibitor （2.5 μmol/L cyclopamine）＋fatty acid group, Shh inhibitor＋fatty acid＋tea polyphenols group and inhibitors of Shh and PARP-1＋ fatty acid ＋tea polyphenols group. The changes of cell viability, apoptosis, nitric oxide （NO） synthesis and oxidative stress related indicators were examined in each group. Results After fatty acid treatment, the survival rate of MS-1 cells was decreased, and the level of apoptosis was significantly increased （P〈0.05）; meanwhile, fatty acid treatment also significantly increased the content of NO, and the activities of total nitric oxide synthase （tNOS）, inducible NOS （iNOS） and constitutive NOS （cNOS） in the cells （P〈0.05）. Moreover, the lipid peroxidation product, malondialdehyde （MDA） was remarkably elevated and the levels of antioxidants, glutathione （GSH） and superoxide dismutase （SOD）, were significantly decreased in response to fatty acid （P〈0.05）; the expression of PARP-1 and phosphorylated Shh （pShh） was significantly increased （P〈0.05）. Tea polyphenols could significantly attenuate the toxic effects of fatty acids concerning all the detected indicators （P〈0.05）. Moreover, after pretreatment with Shh-PARP-1 inhibitors BYK204165 and cyclopamine for 1 h, the protective effects of tea polyphenols were markedly enhanced. There were no significant difference in the detected indicators as compared with c
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2016年第8期975-981,共7页
Academic Journal of Second Military Medical University
