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目的:观察不同浓度的米诺环素对体外培养的髓核细胞-氧化氮合酶(NOS)活性、NO含量、细胞总蛋白含量变化的影响。方法:酶消化及自然传代法分离培养体外兔椎间盘髓核细胞,HE染色观察髓核细胞形态;利用不同浓度(0.1,1,10,100 mg/L)的米诺环素干预P2髓核细胞,分光光度法检测各组培养液中NOS活性和NO含量,考马斯亮蓝法检测各组细胞总蛋白含量。结果:成功建立了兔椎间盘髓核细胞的体外培养模型;米诺环素可以降低NOS活性和减少NO分泌,并且可以增加细胞总蛋白含量,与对照组比较差别具有统计学意义。结论:一定浓度的米诺环素可以有效减缓椎间盘髓核细胞的退变。
OBJECTIVE: To observe the effects of different concentrations of minocycline on the activity of nitric oxide synthase (NOS), NO and total cellular protein in cultured nucleus pulposus cells. Methods: The nucleus pulposus cells were isolated and cultured in vitro by enzymatic digestion and natural passage method. Morphology of nucleus pulposus cells was observed by HE staining. Minocycline (0.1, 1, 10, 100 mg / L) The activity of NOS and the content of NO in each culture medium were detected by spectrophotometry. The content of total protein in each group was detected by Coomassie brilliant blue method. Results: The in vitro culture model of rabbit nucleus pulposus cells was established successfully. Minocycline can reduce the activity of NOS and decrease the secretion of NO, and increase the content of total protein in cells. The difference was statistically significant compared with the control group. Conclusion: A certain concentration of minocycline can effectively reduce the degeneration of nucleus pulposus cells.