高浓度抗生素溶液对椎间盘细胞影响的研究

Influences of the high concentrations of cefazolin and clindamycin on the viability and proliferation of nucleus pulposus cell

[目的]检测高浓度的头孢唑林钠及克林霉素对兔腰椎间盘髓核细胞活力及增殖能力的影响。[方法]通过在体外分离、培养新西兰兔髓核细胞,体外扩增培养至第1代,采用CCK-8及RT-PCR等方法,检测4个浓度(0mg/ml、0.25mg/ml、0.5mg/ml、1mg/ml)的头孢唑啉钠及克林霉素对于兔腰椎间盘髓核细胞活力及增殖能力的影响。[结果]当头孢唑啉钠浓度超过0.5mg/ml、克林霉素浓度超过0.25mg/ml时,髓核细胞Ⅱ型胶原及蛋白多糖的表达下降,浓度在1.0mg/ml时达到最低。头孢唑啉钠浓度超过0.5mg/ml、克林霉素浓度超过0.25mg/ml时,明显影响细胞活力,小于以上浓度时细胞活性无明显影响。[结论]高浓度的头孢唑林钠及克林霉素抗生素对于体外培养的兔髓核细胞的活力、增殖能力具有抑制作用。

[ Objective] To observe the influences of the high concentrations of cefazolin and clindamycin on the viability and proliferation of nucleus pulposus cell. [ Method] NP cells from rabbit disc were harvested and cultured in vitro. Then they were randomly divided into eight groups with different concentrations of cefazolin and clindamycin（0 mg/ml,0.25 mg/ml,0.5 mg/ml, 1mg/ml）. Semi-quantitative real-time polymerase chain reaction （RT-PCR） was used to quantify type Ⅱ collagen and aggrecan respectively. The changes of cells viability were detected by CCK - 8 staining. [ Result] Cefazolin and clindamyein can inhibit type Ⅱ collagen and aggrecan expression at the concentration of exceeding 0.5 mg/ml and 0.25 mg/ml and reached their peaks at 1.0 mg/ml. Cefazolin and clindamycin affected the viability of the NP cells exposed at the concentration of exceeding 0.5 mg/ ml and 0.25 mg/ml. The concentration below the above level didn＇t obviously affect cell activity. [ Conclusionj Cefazolin and clindamycin can inhibit type Ⅱ collagen and aggrecan expression at the concentration of exceeding 0.5 mg/ml and 0.25 mg/ml. Cefazolin and clindamycin affect the viability of the NP cells exposed at the concentration of exceeding 0.5 mg/ml and 0.25mg/ ml.