羧甲基壳聚糖对体外培养许旺细胞增殖及核因子κB表达的影响

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、0，10，50，100，200，500，1 000 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50-1 000 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。

Carboxymethylated chitosan effects on proliferation of Schwann cells and expression of nuclear factor kappa B

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-well plates, and cultured respectively with PBS, 0, 10, 50, 100, 200, 500, 1 000 mg/L carboxymethyl chitosan for 24 hours. Cell proliferation was detected using the cell counting kit-8 assay. After trypsin digestion, Schwann cells from Sprague-Dawley rats at logarithmic growth phase were used to prepare cell suspensions, which were seeded in 6-well cell culture 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： Cell counting kit-8 and 5-bromo-2-deoxyuridine detection results showed that carboxymethyl chitosan at 50-1000 mg/L, especially at 200-500 mg/L, could promote Schwann cell proliferation. Real-time PCR and western blot results showed 50-200 mg/L carboxymethyl chitosan could promote nuclear factor κB mRNA and protein expression in Schwann cells in a dose-dependent manner, suggesting carboxymethyl chitosan can promote Schwann cell proliferation and expression of nuclear factor-κB in Schwann cells cultured in vitro.