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目的 观察促性腺激素释放激素 (gonadotropin releasinghormone,GnRH)类似物阿拉瑞林 (alarelin)对培养的大鼠胃平滑肌细胞增殖的影响。 方法 应用离体培养的SD大鼠胃平滑肌细胞 (gastricsmoothmusclecell,GSMC) ,采用四唑盐 (MTT)比色法实验 ,3H 胸腺嘧啶核苷酸 (3H TdR)参入、免疫荧光化学检测增殖细胞核抗原 (pro liferatingcellnuclearantigen ,PCNA)表达的平均荧光值和流式细胞仪技术 ,观察阿拉瑞林对GSMC的增殖、DNA合成和细胞周期的影响。 结果 当加入终浓度为 10 - 9、10 - 7、10 - 5mol L的阿拉瑞林 2 4h后 ,与未用药的对照组相比 ,发现其MTT吸光度 (A值 )逐渐降低 (P <0 0 5 ) ;PCNA表达的平均荧光值逐渐减弱 (P <0 0 5 ) ;GSMC的3H TdR参入量依次减少 (P <0 0 5 ) ,且药物浓度越大 ,此三者下降越明显。在细胞周期中 ,与对照组相比 ,G1 期细胞所占比例明显增加 (P <0 0 5 ) ;S期细胞所占比例明显减少 ,且随药物浓度的增加而逐渐减少 (P <0 0 5 )。 结论 GnRH类似物可明显抑制大鼠GSMC的增殖作用 ,其作用途径可能是通过平滑肌细胞自身GnRH受体的直接介导而实现的
Objective To observe the effect of alarelin, an analog of gonadotropin releasing hormone (gnRH), on the proliferation of cultured rat gastric smooth muscle cells. Methods Gastric smooth muscle cells (GSMCs) of SD rats were cultured in vitro. MTT colorimetric assay and 3H-thymidine incorporation were used. Immunofluorescence staining was used to detect proliferating cell nuclear antigen ( The average fluorescence value of PCNA expression and flow cytometry were used to observe the effect of Alarelin on the proliferation, DNA synthesis and cell cycle of GSMC. Results Compared with the untreated control group, the MTT absorbance (A value) decreased gradually (P <0 0) after addition of 10 to 9,10 - 7,10 - 5 mol L of Alarelin for 24 hours 5). The mean fluorescence value of PCNA gradually decreased (P <0.05). The 3H TdR incorporation of GSMC decreased in turn (P <0.05), and the higher the drug concentration, the more obvious the decrease of the three. In the cell cycle, the proportion of cells in G1 phase increased significantly (P <0 05) compared with the control group; the proportion of cells in S phase decreased significantly, and decreased with the increase of drug concentration (P <0 0 5). Conclusions GnRH analogue can significantly inhibit the proliferation of rat GSMC, which may be mediated by the direct mediation of GnRH receptor on smooth muscle cells