CAJ | 학술논문

目的：观察槲皮素对高浓度葡萄糖培养的大鼠雪旺细胞系细胞( rat Schwann cell line,RSC96)自噬、增殖活性和凋亡的影响。方法高浓度葡萄糖(125 mmol/L)及槲皮素(25μmol/L)离体培养RSC96细胞,作用72小时后,噻唑基四唑( methyl thiazolyl tetrazolium,MTT)法检测细胞增殖活性,原位末端标记法[ terminal dexynucleotidyl transferase( TdT)-mediated dUTP nick end labe-ling,TUNEL]评价凋亡,Western blot法检测caspase-9活化的P35亚基、caspase-3活化的P20亚基及自噬相关蛋白Beclin1的蛋白表达。结果高糖抑制离体培养的RSC96细胞的增殖活性,吸光度(optical density,OD)值降低,凋亡率增加,P35亚基、P20亚基蛋白表达上调(P<0．05,P<0．01)；槲皮素能够增加高糖培养RSC96细胞的增殖活性,下调P35、P20蛋白表达及凋亡率( P<0．05,P<0．01)。然而当自噬被抑制后,槲皮素上述保护作用则被削弱,细胞死亡增多(P<0．01)。结论槲皮素通过增加自噬以促进RSC96细胞的增殖、减少凋亡,从而减轻高浓葡萄糖的损伤。
목적：관찰곡피소대고농도포도당배양적대서설왕세포계세포( rat Schwann cell line,RSC96)자서、증식활성화조망적영향。방법고농도포도당(125 mmol/L)급곡피소(25μmol/L)리체배양RSC96세포,작용72소시후,새서기사서( methyl thiazolyl tetrazolium,MTT)법검측세포증식활성,원위말단표기법[ terminal dexynucleotidyl transferase( TdT)-mediated dUTP nick end labe-ling,TUNEL]평개조망,Western blot법검측caspase-9활화적P35아기、caspase-3활화적P20아기급자서상관단백Beclin1적단백표체。결과고당억제리체배양적RSC96세포적증식활성,흡광도(optical density,OD)치강저,조망솔증가,P35아기、P20아기단백표체상조(P<0．05,P<0．01)；곡피소능구증가고당배양RSC96세포적증식활성,하조P35、P20단백표체급조망솔( P<0．05,P<0．01)。연이당자서피억제후,곡피소상술보호작용칙피삭약,세포사망증다(P<0．01)。결론곡피소통과증가자서이촉진RSC96세포적증식、감소조망,종이감경고농포도당적손상。
Objective To observe the protective effect of quercetin on the RSC96 cells cultured in high glucose in the aspects of autophagy, proliferation and apoptosis. Methods RSC96 cells were cultured with high glucose or quercetin media. The proliferation was detected by MTT method;the apoptosis was de-tected by TUNEL method;the expression of the activated subunit P35 of caspase-9 and the activated sub-unit P20 of caspase-3 were detected by Western blot method. Results The increased concentration of glu-cose lowered the proliferation of RSC96 cells and increased the apoptosis (P<0. 05, P<0. 01). Quercetin significantly up-regulated the proliferation rate, and down-regulated the apoptosis by attenuated the high glucose-induced activation of caspase-3 and caspase-9 (P<0. 05, P<0. 01). Furthermore, inhibition of autophagy led to significantly attenuate the protective effect of quercetin (P<0. 01). Conclusions Quer-cetin decreased apoptosis and up-regulated proliferation on the RSC96 cells cultured in high glucose through the autophagy pathway.