人参皂苷Rb1抑制间歇性高糖诱导的雪旺细胞凋亡

Ginsenoside Rb1 inhibits apoptosis in Schwann cells induced by intermittent high glucose

目的探讨人参皂苷Rb1对间歇性高糖培养条件下雪旺细胞凋亡的影响。方法无菌剥取新生Sprague-Dawley乳鼠坐骨神经,体外制备雪旺细胞,分为对照组(培养液葡萄糖浓度为5.6 mmol/L,Con组)、间歇性高糖组(培养液葡萄糖浓度为5.6 mmol/L和50 mmol/L,每8小时交替,IHG组)、干预组(在间歇性高糖基础上加入100μmol/L人参皂苷Rb1,IHG+Rb1组)分别培养48 h。采用Annexin V/PI双染色、流式细胞仪检测细胞凋亡,ELISA法检测8-羟基脱氧鸟苷(8-OHd G)水平,实时荧光定量PCR法检测凋亡相关基因Bcl-2和Bax的mRNA表达,蛋白印迹法检测Bcl-2和Bax的蛋白表达。结果与对照组相比,间歇性高糖促进了雪旺细胞凋亡(P<0.05),增加了8-OHd G的生成(P<0.01),IHG组细胞凋亡率和8-OHd G水平分别是Con组的7.69倍、3.24倍;间歇性高糖上调了Bax的mRNA和蛋白表达,下调了Bcl-2的mRNA和蛋白表达。与IHG组相比,人参皂苷Rb1抑制了IHG导致的雪旺细胞凋亡和8-OHd G生成,IHG+Rb1组细胞凋亡率和8-OHd G水平较IHG组分别下降了54.1%和32.7%(P<0.05);同时下调了Bax的mRNA和蛋白表达,上调了Bcl-2的mRNA和蛋白表达(P<0.05)。结论间歇性高糖可促进雪旺细胞凋亡,人参皂苷Rb1能够减轻间歇性高糖造成的凋亡,具有一定的神经保护作用。

Objective To investigate the effects of ginsenoside Rbl on the apoptosis in Schwann cells induced by intermittent high glucose（IHG）.Methods Schwann cells were obtained and primarily cultured from the sciatic nerves of neonatal Sprague-Dawley rats.The cultured cells were treated for 48 h with 5.6 mmol/L glucose as control（Con group） pr with 5.6 mmol/L and 50 mmol/L glucose alternating every 8 h to mimic IHG（IHG group）,or with IHG plus 100 μmol/L ginsenoside Rb1（IHG＋Rb1 group）.Apoptosis was detected by flow cytometry.Level of 8-hy—droxy-2-deoxy guanosine（8-OHdG） was detected by enzyme linked immunosorbent assay.The mRNA expression of Bax and Bcl-2 was detected by quantitative real-time reverse transcription PCR.The protien expression of Bax and Bcl—2 was detected by Western blot.Results As compared with Con group,the percentage of apoptotic cells and the level of 8-OHdG in IHG group were both increased（7.69 and 3.24 folds of control respectively,P 〈0.01）.The mRNA and protein expression of Bax were up-regulated,while those of Bcl-2 were down-regulated in IHG group.Rbl treatment inhibited the above effects induced by IHG.The percentage of apoptotic cells and the level of 8-OHdG in IHG＋Rb1group were decreased by 54.1%and 32.7%respectively as compared with IHG group（P 〈0.05）.The down-regula—ted mRNA and protein expression of Bax,and the up-regulated mRNA and protein expression of Bcl-2 were antagonized in IHG＋Rb1 group as compared with IHG group.Conclusion Ginsenoside Rb1 inhibits IHG-induced apoptosis in Schwann cells with certain neuroprotective effect