CAJ | 학술논문

背景：研究表明羧甲基壳聚糖对多种细胞具有促增殖作用，但其对许旺细胞增殖的影响及具体作用机制有待进一步探索。 　　目的：观察羧甲基壳聚糖对许旺细胞的促增殖作用，以及其对许旺细胞内核因子κB表达及活性的影响。 　　方法：取对数生长期的SD乳鼠许旺细胞细胞悬液接种于96孔板，分别以PBS、0，10，50，100，200，500，1000 mg/L羧甲基壳聚糖培养24 h，采用CCK-8法检测细胞增殖。取对数生长期的SD乳鼠许旺细胞，胰酶消化后制备成细胞悬液，接种于6孔细胞培养板内，分别加入50 mg/L羧甲基壳聚糖、100 mg/L羧甲基壳聚糖、200 mg/L羧甲基壳聚糖、PBS培养24 h，进行BrdU、Real-time PCR及Western blot法检测。结果与结论：CCK-8及 BrdU 检测结果表明羧甲基壳聚糖在50-1000 mg/L 范围内可促进许旺细胞增殖，200-500 mg/L时促增殖效应最明显。Real-time PCR及Western blot结果表明50-200 mg/L羧甲基壳聚糖可促进许旺细胞内核因子κB mRNA及蛋白的表达，且具有浓度依赖性。表明羧甲基壳聚糖可促进体外培养许旺细胞的增殖及其核因子κB的表达。
배경：연구표명최갑기각취당대다충세포구유촉증식작용，단기대허왕세포증식적영향급구체작용궤제유대진일보탐색。 　　목적：관찰최갑기각취당대허왕세포적촉증식작용，이급기대허왕세포내핵인자κB표체급활성적영향。 　　방법：취대수생장기적SD유서허왕세포세포현액접충우96공판，분별이PBS、0，10，50，100，200，500，1000 mg/L최갑기각취당배양24 h，채용CCK-8법검측세포증식。취대수생장기적SD유서허왕세포，이매소화후제비성세포현액，접충우6공세포배양판내，분별가입50 mg/L최갑기각취당、100 mg/L최갑기각취당、200 mg/L최갑기각취당、PBS배양24 h，진행BrdU、Real-time PCR급Western blot법검측。결과여결론：CCK-8급 BrdU 검측결과표명최갑기각취당재50-1000 mg/L 범위내가촉진허왕세포증식，200-500 mg/L시촉증식효응최명현。Real-time PCR급Western blot결과표명50-200 mg/L최갑기각취당가촉진허왕세포내핵인자κB mRNA급단백적표체，차구유농도의뢰성。표명최갑기각취당가촉진체외배양허왕세포적증식급기핵인자κB적표체。
BACKGROUND:Carboxymethylated chitosan is shown to promote some kinds of cells proliferation, but its effects on proliferation of Schwann cells need further studies. OBJECTIVE:To investigate the effects of carboxymethylated chitosan on proliferation of Schwann cells and expression of nuclear factor-κB in cultured Schwann cells. METHODS:Schwann cells from Sprague-Dawley rats at logarithmic growth phase were seeded in 96-wel plates, and cultured respectively with PBS, 0, 10, 50, 100, 200, 500, 1 000 mg/L carboxymethyl chitosan for 24 hours. cellproliferation was detected using the cellcounting kit-8 assay. After trypsin digestion, Schwann cells from Sprague-Dawley rats at logarithmic growth phase were used to prepare cellsuspensions, which were seeded in 6-wel cellculture plates and cultured respectively with 50, 100 and 200 mg/L carboxymethyl chitosan and PBS for 24 hours. Then, 5-bromo-2-deoxyuridine, real-time PCR and western blot assay were performed. RESULTS AND CONCLUSION:cellcounting kit-8 and 5-bromo-2-deoxyuridine detection results showed that carboxymethyl chitosan at 50-1000 mg/L, especial y at 200-500 mg/L, could promote Schwann cellproliferation. Real-time PCR and western blot results showed 50-200 mg/L carboxymethyl chitosan could promote nuclear factorκB mRNA and protein expression in Schwann cells in a dose-dependent manner, suggesting carboxymethyl chitosan can promote Schwann cellproliferation and expression of nuclear factor-κB in Schwann cells cultured in vitro.