孕酮对子宫蓄脓犬子宫内膜基质细胞增殖的影响

Effect of Progesterone on Proliferation of Canine Endometrial Stromal Cells with Pyometra

旨在研究子宫蓄脓对犬子宫内膜基质细胞(endometrial stromal cells,ESCs)增殖的影响及孕酮(progesterone,P_(4))的干预作用。收集健康犬和经腹部B超确诊患子宫蓄脓犬的子宫,采用胶原酶消化法分离ESCs,并通过细胞免疫荧光检测进行鉴定;采用CCK-8法检测培养不同时间后两种ESCs的细胞活力及孕酮作用的影响。结果显示,胶原酶消化法可分离获得高纯度的犬ESCs;将分离的ESCs培养24 h和48 h时,子宫蓄脓犬ESCs的细胞活力显著高于健康犬(P<0.01)。用0、5、15、30 ng/mL等不同浓度P_(4)处理两种ESCs,当作用24 h时,两种细胞的活力与相应的对照比均无显著变化(P>0.05);作用48 h时,30 ng/mL的P_(4)可显著促进健康犬ESCs的增殖(P<0.01),而对子宫蓄脓犬ESCs的增殖具有显著的抑制作用(P<0.05);作用72 h时,3种浓度P_(4)均显著促进健康犬ESCs的增殖(P<0.01),但对子宫蓄脓犬ESCs的增殖均无显著影响(P>0.05)。以上结果表明,子宫蓄脓犬ESCs的快速增殖可能与子宫内膜的囊性增生密切相关,P_(4)可促进健康犬ESCs的增殖,但在一定程度上抑制子宫蓄脓犬ESCs的增殖。

This study was aimed to investigate the effect of pyometra on proliferation of canine endometrial stromal cells(ESCs)as well the effect of progesterone.Uteri were collected from clinically healthy dogs and dogs with pyometra diagnosed by abdominal B-ultrasound.ESCs were isolated by collagenase digestion and identified by cellular immunofluorescence assay.CCK-8 assay was performed to detect the cell viability of the two types of ESCs after different culture times or progesterone treatment.The results showed that canine ESCs with high purity were obtained by collagenase digestion;The viability of ESCs from pyogenic dogs was significantly higher than that from healthy ones(P<0.01)after the isolated ESCs were cultured for 24 h and 48 h.When the ESCs were treated with different concentrations of progesterone(0 ng/mL,5 ng/mL,15 ng/mL and 30 ng/mL)for 24 h,the viability of these two types of cells showed no difference compared with the corresponding controls(P>0.05);At 48 h of treatment,30 ng/mL progesterone significantly promoted the proliferation of ESCs in healthy dogs(P<0.01),but significantly inhibited the proliferation of the canine ESCs with pyometra(P<0.05);At 72 h,treatment with the three concentrations of progesterone significantly increased the proliferation of ESCs in healthy dogs(P<0.01),but no significant difference was obtained with pyogenic ESCs(P>0.05).The data indicate that rapid proliferation of ESCs may be closely related to the cystic endometrial hyperplasia in canine pyometra;P_(4) promotes the proliferation of ESCs in clinically healthy dogs,but to a certain extent inhibits the proliferation of ESCs in pyogenic dogs.