CAJ | 학술논문

背景：研究发现吡咯喹啉醌可以促进许旺细胞增殖及生长因子的分泌，但其对关节软骨或关节软骨细胞有何作用尚未见报道。 　　目的：验证吡咯喹啉醌对膝关节软骨细胞增殖及对白细胞介素1β介导的软骨细胞凋亡的影响，以探讨吡咯喹啉醌保护软骨细胞的作用机制。 　　方法：在无菌环境下消化得到1月龄新西兰白兔膝关节软骨细胞，培养并传代，第2代软骨细胞用于实验。细胞贴壁后用吡咯喹啉醌浓度为0，6.25，12.5，25.0，50.0，100.0μmol/L的无血清培养基分别培养软骨细胞48 h，采用MTT法检测细胞的增殖活力；培养30 h，采用流式细胞术测细胞周期。细胞贴壁后用不同浓度吡咯喹啉醌预处理软骨细胞24 h，然后加入白细胞介素1β作用15 h后，采用流式细胞术测细胞凋亡率。 　　结果与结论：吡咯喹啉醌能明显提高软骨细胞的增殖活力、S期，G2/M比例和细胞增殖指数(P <0.05)，且在吡咯喹啉醌浓度为12.5μmol/L和25.0μmol/L时作用最强。吡咯喹啉醌能明显抑制白细胞介素1β介导的软骨细胞早期凋亡和晚期凋亡(P <0.05)，在吡咯喹啉醌浓度为25.0μmol/L时作用最明显。结果表明吡咯喹啉醌可以促进膝关节软骨细胞的分裂与增殖，对白细胞介素1β介导的软骨细胞凋亡有抑制作用。
배경：연구발현필각규람곤가이촉진허왕세포증식급생장인자적분비，단기대관절연골혹관절연골세포유하작용상미견보도。 　　목적：험증필각규람곤대슬관절연골세포증식급대백세포개소1β개도적연골세포조망적영향，이탐토필각규람곤보호연골세포적작용궤제。 　　방법：재무균배경하소화득도1월령신서란백토슬관절연골세포，배양병전대，제2대연골세포용우실험。세포첩벽후용필각규람곤농도위0，6.25，12.5，25.0，50.0，100.0μmol/L적무혈청배양기분별배양연골세포48 h，채용MTT법검측세포적증식활력；배양30 h，채용류식세포술측세포주기。세포첩벽후용불동농도필각규람곤예처리연골세포24 h，연후가입백세포개소1β작용15 h후，채용류식세포술측세포조망솔。 　　결과여결론：필각규람곤능명현제고연골세포적증식활력、S기，G2/M비례화세포증식지수(P <0.05)，차재필각규람곤농도위12.5μmol/L화25.0μmol/L시작용최강。필각규람곤능명현억제백세포개소1β개도적연골세포조기조망화만기조망(P <0.05)，재필각규람곤농도위25.0μmol/L시작용최명현。결과표명필각규람곤가이촉진슬관절연골세포적분렬여증식，대백세포개소1β개도적연골세포조망유억제작용。
BACKGROUND:Pyrroloquinoline quinone is found to accelerate Schwann cel proliferation and growth factor secretion, but there is no report addressing its role in articular cartilage and chondrocytes. OBJECTIVE: To investigate the role of pyrroloquinoline quinone in chondrocyte proliferation and interleukin-1β-induced chondrocyte apoptosis in the articular cartilage of knee joints and to verify the protective mechanism involved. METHODS: Chondrocytes were isolated from New Zealand white rabbits (1 month of age), digested under aseptic conditions, and cultured in DMEM/F12 in the presence of 10% fetal bovine serum to alow for proliferation until passage 2. Adherent chondrocytes were cultured in serum-free DMEM/F12 medium with 0, 6.25, 12.5, 25.0, 50.0 and 100.0 μmol/L pyrroloquinoline quinone, separately. Proliferation activity was determined by MTT at 48 hours of pyrroloquinoline quinone administration. Cel cycle was determined by flow cytometry at 30 hours after pyrroloquinoline quinone administration. Apoptosis was determined by flow cytometry folowing 24 hours of pyrroloquinoline quinone pretreatment and 15 hours of interleukin-1β induction. RESULTS AND CONCLUSION: Pyrroloquinoline quinone enhanced chondrocyte proliferation activity, increased percentage of S phase and G2/M phase in a dose dependent manner and reached the peak when the concentration of pyrroloquinoline quinone was 12.5-25.0 μmol/L (P< 0.05). Pyrroloquinoline quinone also inhibited interleukin-1β-induced chondrocyte apoptosis in early and late stage, and 25.0 μmol/L pyrroloquinoline quinone had the best effects (P < 0.05). These findings suggest pyrroloquinoline quinone can promote chondrocyte division and proliferation, and protect the cels from interleukin-1β-induced apoptosis.