目的 探討補(bǔ)骨脂素對(duì)H₂O₂誘導(dǎo)PC-12細(xì)胞氧化損傷的保護(hù)作用,并考察其作用機(jī)制。方法 PC-12細(xì)胞分為對(duì)照組、模型組、Tempol組和補(bǔ)骨脂素組。對(duì)照組中未加入補(bǔ)骨脂素和H₂O₂;模型組中加入400μmol/LH₂O₂;Tempol組中加入100μmol/LTempol及400μmol/LH₂O₂;補(bǔ)骨脂素組中加入400μmol/LH₂O₂及1、5、10、20μmol/L補(bǔ)骨脂素。CCK-8法測(cè)定PC-12細(xì)胞的存活率,考察對(duì)PC-12細(xì)胞形態(tài)的影響,PI/Annexin-V、Hoechst染色檢測(cè)細(xì)胞凋亡率,蛋白質(zhì)印跡法檢測(cè)凋亡蛋白表達(dá)。結(jié)果 5、10、20μmol/L補(bǔ)骨脂素均可以顯著提高PC-12細(xì)胞內(nèi)的吸光度值(P<0.01),細(xì)胞存活率提高較明顯,與模型組比較差異具有統(tǒng)計(jì)學(xué)意義(P<0.05)。補(bǔ)骨脂素1、5、10、20μmol/L組凋亡率明顯低于模型組(P<0.05),并且隨著補(bǔ)骨脂素濃度的增加,PC-12細(xì)胞的凋亡率逐漸下降。隨著補(bǔ)骨脂素作用時(shí)間的增加,各組PC-12細(xì)胞凋亡率的增加幅度相近,但明顯低于模型組(P<0.05)。隨著補(bǔ)骨脂素濃度的增加,對(duì)磷酸化Bad、Bcl-XL水平的上調(diào)作用越明顯。結(jié)論 補(bǔ)骨脂素對(duì)H₂O₂誘導(dǎo)PC-12細(xì)胞損傷具有保護(hù)作用,其作用機(jī)制是通過(guò)上調(diào)磷酸化Bad和Bcl-XL表達(dá)來(lái)抑制細(xì)胞凋亡。

Objective To discuss the protective effect of psoralen on PC-12 cell damage induced by H₂O₂, and investigate its mechanism. Methods PC-12 cells were divided into control model, Tempol, and psoralen groups. Cells in the control group did not receive damage or treatments, the model group was established by 400 μmol/L H₂O₂-damage, Tempol group was cultured by 100 μmol/L Tempol and 400 μmol/L H₂O₂, and psoralen groups were cultured by 400 μmol/L H₂O₂ and 1, 5, 10, and 20 μmol/L psoralen. Cell survival rates in four groups were detected by CCK-8 method, apoptosis rates were detected by PI/Annexin and Hoechst methods, and protein expression were detected by Western blotting. Results Compared with the model group, psoralen (1, 5, 10, and 20 μmol/L) could significantly improve absorbance values (P < 0.01) and cell survival rates (P < 0.05) of PC-12 cells. Apoptosis rates of psoralen (1, 5, 10, and 20 μmol/L) groups were significantly lower than those of model group (P < 0.05). And with increase of psoralen concentration, apoptosis rates of PC-12 cells were gradually decreased. With longer action time, the increase amplitudes of PC-12 cells apoptosis rates among groups were similar, and obviously lower than those of model group (P < 0.05). Psoralen could significantly up-regulate levels of p-Bad and Bcl-XL with increase of concentrations. Conclusion Psoralen has protective effect on PC-12 cell damage induced by H₂O₂, which is related to up-regulation of p-Bad and Bcl-XL to inhibit the apoptosis of PC-12 cell.