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目的:探讨甲基强的松龙(MP)对体外培养许旺氏细胞的作用。方法:取成年SD大鼠坐骨神经以植块法分离纯化许旺氏细胞,将纯化后的细胞培养2周后分为5组移入96孔板,A、B、C组为MP组(按加入药物浓度不同分组20μg/ml,2μg/ml,0.2μg/ml),D组为神经生长因子(NGF)组(药物浓度0.1μg/ml),E组为对照组。继续培养9d后倒置显微镜下观察细胞生长情况并以MTT法测定各组许旺氏细胞的存活与增殖能力。结果:倒置显微镜下观察小剂量MP组及NGF组细胞生长优于其他各组。570nm波长分光光度计测定A值,小剂量(0.2μg/ml)MP组为0.376±0.079,中剂量(2μg/ml)MP组0.286±0.116,大剂量(20μg/ml)MP组0.280±0.086,NGF组0.320±0.121,对照组0.252±0.115,小剂量MP组明显优于大剂量MP组(P<0.05),同时优于对照组(P<0.05),余各组间均无明显统计学差异。结论:小剂量MP能够促进体外培养许旺氏细胞增殖,从而促进周围神经损伤的修复,细胞培养用药浓度为0.2μg/ml。MP浓度过大(相当于临床冲击剂量的血峰浓度)将抑制细胞增殖。
Objective: To investigate the effect of methylprednisolone (MP) on Schwann cells cultured in vitro. METHODS: Schwann cells were isolated and purified from adult sciatic nerve of SD rats. The purified cells were divided into five groups and then transplanted into 96-well plates after two weeks. Groups A, B and C were treated with MP (20μg / ml, 2μg / ml, 0.2μg / ml) with different concentrations. The rats in group D were treated with nerve growth factor (NGF) (0.1μg / ml). Continue to cultivate 9d after inverted microscope to observe the cell growth and MTT method to determine the survival and proliferation of Schwann cells in each group. Results: Under inverted microscope, the cell growth of MP group and NGF group was better than other groups. A value of 0.376 ± 0.079 in low dose (0.2μg / ml) MP group, 0.286 ± 0.116 in medium dose (2μg / ml) MP group and 0.280 ± 0.086 in high dose (20μg / ml) MP group by 570nm wavelength spectrophotometer, 0.320 ± 0.121 in the NGF group and 0.252 ± 0.115 in the control group were significantly better than those in the high-dose MP group (P <0.05), and were superior to the control group (P <0.05). There was no significant difference between the other groups . CONCLUSION: Small doses of MP can promote the proliferation of Schwann cells cultured in vitro and promote the repair of peripheral nerve injury. The concentration of MP in cell culture is 0.2μg / ml. MP concentration is too large (equivalent to the clinical impact dose peak concentration) will inhibit cell proliferation.