人参皂甙Rg1对压力性尿失禁患者尿道旁筋膜成纤维细胞增殖的影响

Effect of ginsenoside-Rgl on the proliferation of paraurethral fascia fibroblasts derived from women suffering from stress urinary incontinence

目的研究人参皂甙Rg1对体外培养的压力性尿失禁（SUI）患者的尿道旁筋膜成纤维细胞增殖的影响。方法选择2006-2007年行尿道中段悬吊术的SUI患者术中切取的阴道前壁组织4份，剔除黏膜层，组织块培养法于体外培养尿道旁筋膜成纤维细胞，胰蛋白酶消化传代3～5代后，将细胞分为4组，分别加入5、10、20μmol／L人参皂甙Rg1及培养液，采用四甲基偶氮唑蓝比色法检测不同浓度人参皂甙Rg1培养24、48、72h后的细胞增殖率，采用免疫组织化学法检测人参皂甙Rg1培养48h后增殖细胞核抗原（PCNA）的表达。结果（1）加入5、10、20μmol／L人参皂甙Rg1培养72h后，各组的细胞增殖率分别为（294-5）％、（40±5）％、（26±4）％，分别与对照组（0）比较，差异均有统计学意义（P〈0．01）；10μmol／L组分别与5txmol／L组、20μmol／L组比较，差异也均有统计学意义（P〈0．01）；但5Ixmol／L组与20μmol／L组比较，差异无统计学意义（P〉0．05）。（2）同一浓度人参皂甙Rg1组培养48h后的细胞增殖率与培养24h时比较，差异均有统计学意义（P〈0．01）；培养72h后与培养48h时比较，差异也均有统计学意义（P〈0．01）。（3）5μmol／L组、10μmol／L组及20μmol／L组培养48h后的PCNA阳性率分别为49．24％、83．48％和54．50％，分别与对照组（28．77％）比较，差异均有统计学意义（P〈0．01）；10μmol／L组与5μmol／L组、20μmol／L组分别比较，差异也有统计学意义（P〈0．01）。结论人参皂甙Rg1可促进体外培养的SUI患者尿道旁筋膜成纤维细胞的增殖。

Objective To investigate the effect of ginsenoside-Rg1 on paraurethral fascia fibroblasts multiplication and the expression of proliferation cell nuclear antigen （PCNA） of stress urinary incontinence （SUI） women in vitro. Methods Specimens of human paraurethral fascia were obtained from 4 SUI women during tension-free vaginal tape （TVT） or tension-free vaginal tape-obturator （TVT-O） procedure. Fibroblasts were isolated and cultured by outgrowth technique. After reaching confluency fibroblasts were subcultured every 5 days and cells after passage number 3 to 5 were used for assessment. The paraurethral fascia fibroblasts were treated with ginsenoside-Rg1 at different concentrations （5, 10, 20 μmol/L） and fibroblasts without Rg1 were used as control. The multiplication conditions of paranrethral fascia fibroblasts were respectively detected by methyl thiazolyl tetrazolium （MTF） assay and the expression of PCNA by histochemistry. Results （ 1 ） Compared with the control group, the growth rate of cells treated with different concentrations of Rg1 after 72h [ （29 ±5）%, （40 ±5） %, （26 ±4） % respectively] was significantly higher （P 〈 0. 01 ）. （2）Compared with the control group, the stimulatory effect of Rg1 on fibroblast growth was significant at 24 h （ P 〈 0. 01 ）, and peaked at 72 h [ （ 29 ± 5 ）%, （ 40 ± 5 ）%, （ 26 ± 4 ） % respectively, P 〈 0. 01 ]. （3） Compared with the control group（28.77% ）, there was a significant increase of PCNA-positive cells （ P 〈 0. 01 ） after 48 h treatment with different concentrations of Rg1 （49. 24%, 83.48%, 54. 50% respectively）. Conclusion The results indicate that, at least in vitro, fibroblasts from paraurethral fascia taken from women suffering from SUI are able to proliferate after ginsenoside-Rg1 treatment.