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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 regulatory effect of Sonic Hedgehog (Shh) - poly (ADP-ribose) polymerase 1 (PARP-1) signaling pathway on the lipotoxicity of tea polyphenols against islet microvascular endothelial cells. Methods Mouse islet microvascular endothelium MS-1 cells were divided into normal control group, solvent control group, fatty acid (0.25 mmol / L palmitate + 0.5 mmol / L oleic acid) L group, fatty acid + tea polyphenol group, PARP-1 inhibitor (8μmol / L BYK204165) + fatty acid group, PARP-1 inhibitor + fatty acid + tea polyphenol group, Shh inhibitor (2.5μmol / ) + Fatty acid group, Shh inhibitor + fatty acid + tea polyphenols group and Shh inhibitor + PARP-1 inhibitor + fatty acid + tea polyphenol group, the cell viability, the level of apoptosis, the levels of NO, Synthesis and changes related to oxidative stress. Results After fatty acid treatment, the survival rate of MS-1 cells decreased and the apoptosis rate increased (P <0.05). Meanwhile, the content of NO and the content of total nitric oxide synthase (tNOS), inducible NOS (iNOS) (P <0.05). Moreover, the content of malondialdehyde (MDA) in lipid peroxidation products increased (P <0.05), the content of antioxidant glutathione (GSH) and superoxide The level of SOD decreased (P <0.05) and the expression of PARP-1 and phosphorylated Shh increased (P <0.05). After the intervention of tea polyphenols, the level of each index was improved (P <0.05). Moreover, the antagonistic effect of tea polyphenols on fatty acids was more obvious after pretreatment with BYK204165 and cyclopamine for 1 h. The detection indexes The difference between the normal control group was not statistically significant (P> 0.05). CONCLUSION: Fatty acid can induce endothelial dysfunction of islets, and tea polyphenols can antagonize fatty acid toxicity. Inhibition of Shh-PARP-1 signal pathway can enhance the protective effect of tea polyphenols.