摘要
目的观察活血化瘀中药对子宫肌瘤Wnt信号转导通路抑制因子表达的影响。方法采用胶原酶I消化法无菌取得子宫肌瘤细胞/肌细胞并进行传代。分别给予不同浓度的活血化瘀中药处理,CCK8法测定细胞生长抑制率;荧光定量RT-PCR法检测子宫肌瘤细胞Wnt信号转导通路的抑制因子SFRP1表达;免疫细胞化学Western blot法检测SFRP1蛋白表达变化。结果①原代培养的子宫肌瘤细胞呈梭形或者椭圆形,呈束的平滑肌细胞平行排列,部分区域以峰谷样生长,经免疫组化鉴定见a-actin染色表达为阳性。②使用不同浓度的活血化瘀中药24、48、72 h后,可见其对子宫肌瘤细胞有明显抑制作用,且与剂量、时间呈正相关。③人子宫肌瘤细胞SFRP1 mRNA及蛋白含量明显高于人正常子宫肌层细胞,活血化瘀中药干预子宫肌瘤细胞48 h后,SFRP1mRNA及蛋白表达均下调。结论活血化瘀中药可抑制体外培养的人子宫肌瘤细胞增殖。随着药物剂量的增加和用药时间的延长,对子宫肌瘤细胞的增殖作用增强,呈剂量和时间依赖性。活血化瘀中药通过影响Wnt抑制因子SFRP1的表达抑制子宫肌瘤细胞的增生。影响Wnt信号转导通路的抑制因子的表达是可能活血化瘀中药治疗子宫肌瘤的重要机制之一。
Objective To evaluate the effect of blood-activiating and stasis-removing Chinese drugs on the Wnt signaling Pathway inhibitory factors of human uterine leiomyoma cell and explore whether blood-activiating and stasis-removing Chinese drugs can inhibits human uterine leiomyoma cell by Wnt signaling pathway inhibitory factors and then demonstrate that Wnt signaling pathway is one of the target of treating human uterine leiomyoma.Methods Uterine fibroid cells/muscle cells were aseptically obtained using collagenase I digestion method and passaged.And then the cells were treated with different concentrations of traditional Chinese medicine for activating blood circulation and removing blood stasis.The inhibitory effect on the cells was investigated by CCK-8.The expression of Wnt signaling pathway inhibitory factor SFRP1 in uterine leiomyoma cells was detected by fluorescence quantitative RT-PCR.Immunocytochemical Western blot detected the SFRP1 protein expression changes.Result①The primary cultured uterine fibroid cells were spindle shaped or oval shaped,with bundles of smooth muscle cells arranged in parallel,and some areas growing in a peak valley pattern.Immunohistochemical identification showed positive expression of a-actin staining.②After using different concentrations of Chinese drugs for promoting blood circulation and removing blood stasis for 24,48 and 72 hours,it had a significant inhibitory effect on uterine fibroid cells,and was positively correlated with dosage and time.③The mRNA and protein expressions of SFRP1 in human uterine fibroid cells were significantly higher than those in normal uterine myometrial cells.After 48 hours of intervention with traditional Chinese medicine for promoting blood circulation and removing blood stasis,the expressions of SFRP1 mRNA and protein were down-regulated in uterine fibroid cells.Conclusion Chinese drugs for promoting blood circulation and removing blood stasis can inhibit the proliferation of human uterine fibroid cells cultured in vitro.With the increase of drug dosage and the prolongation of medication time,the proliferative effect on uterine fibroid cells is enhanced in a dose-and time-dependent manner.Chinese drugs for promoting blood circulation and removing blood stasis inhibit the proliferation of uterine fibroid cells by affecting the expression of Wnt inhibitory factor SFRP1.The expressions of inhibitory factors affecting the Wnt signaling pathway may be one of the important mechanisms of Chinese drugs for promoting blood circulation and removing blood stasis in the treatment of uterine leiomyoma.
作者
吴中恺
张婷婷
史杨
赵莉
WU Zhongkai;ZHANG Tingting;SHI Yang;ZHAO Li(Yueyang Hospital of Integrated Chinese and Western Medicine Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200437,China;Nanyang Central Hospital,Nanyang 473000,Henan,China)
出处
《辽宁中医杂志》
北大核心
2026年第2期154-156,I0004,共4页
Liaoning Journal of Traditional Chinese Medicine
基金
国家自然科学基金面上项目(82074475)
上海市卫健委名老中医学术经验研究工作室(SHGZS-202229)。
关键词
活血化瘀中药
子宫肌瘤
Wnt信号转导通路抑制因子
SFRP1
blood-activiating and stasis-removing Chinese drugs
uterine leiomyoma
Wnt signaling pathway inhibitory factor
SFRP1
