CAJ | 학술논문

目的良性前列腺增生(BPH)是严重危害老年男性健康的常见疾病,本研究旨在研究组织因子途径抑制因子(TFPI)基因对前列腺平滑肌细胞生长的影响,为良性前列腺增生的基因治疗提供参考依据.方法取前列腺增生患者手术切除的前列腺组织,采用酶消化法分离前列腺平滑肌细胞;免疫组化方法进行细胞鉴定;pIRES-TFPI基因转染前列腺平滑肌细胞,以pIRES基因作为基因转染阴性对照;用细胞计数法和四氮唑蓝MTT法观察细胞增殖情况;采用RT-PCR检测细胞内TFPI基因的表达情况.结果 SMA免疫组化染色和MASSON染色显示:经过5次传代后,前列腺平滑肌细胞的纯度达到95%以上;TFPI基因转染后,前列腺平滑肌细胞内的TFPI mRNA水平明显提高,是未转染组的7倍、阴性对照基因转染组的3.5倍;基因转染4 d后,TFPI基因转染组的前列腺平滑肌细胞数明显低于阴性对照基因转染组.结论提示TFPI基因对前列腺平滑肌细胞的增殖具有调控作用,有必要对其作用机理进行进一步研究.
목적 량성전렬선증생(BPH)시엄중위해노년남성건강적상견질병,본연구지재연구조직인자도경억제인자(TFPI)기인대전렬선평활기세포생장적영향,위량성전렬선증생적기인치료제공삼고의거.방법취전렬선증생환자수술절제적전렬선조직,채용매소화법분리전렬선평활기세포;면역조화방법진행세포감정;pIRES-TFPI기인전염전렬선평활기세포,이pIRES기인작위기인전염음성대조;용세포계수법화사담서람MTT법관찰세포증식정황;채용RT-PCR검측세포내TFPI기인적표체정황.결과 SMA면역조화염색화MASSON염색현시:경과5차전대후,전렬선평활기세포적순도체도95%이상;TFPI기인전염후,전렬선평활기세포내적TFPI mRNA수평명현제고,시미전염조적7배、음성대조기인전염조적3.5배;기인전염4 d후,TFPI기인전염조적전렬선평활기세포수명현저우음성대조기인전염조.결론 제시TFPI기인대전렬선평활기세포적증식구유조공작용,유필요대기작용궤리진행진일보연구.
Objective To investigate the effect of tissue factor pathway inhibitor(TFPI) gene on the proliferation of the prostatic smooth muscle cells in hope for exploring alternative therapeutic approaches for benign prostatic hyperplasia (BPH). Methods Prostatic smooth muscle cells were isolated form the prostatic tissue of the BPH patients by enzymatic digestion assay. Immunohistochemical staining was employed for the confirmation of the prostatic: smooth muscle cells. The prostatic smooth muscle cells were transfeeted with either pIRES-TFPI or pIRES plasmid DNA. MTT assay and cell number counting were employed for the assessment of the proliferation of the cells. Reverse transcription polymerase chain reaction(RT-PCR) was employed for the quantification of TFPI mRNA expression in the cells. Results Immunohistoehemical staining showed that the prostaic smooth muscle cells accounted for about 95 percent of the overall cells at passage 5. The TFPI mRNA level in the cells transfected with the pIRES-TFPI plasmid was 6 fold higher than that in the untreated cells and 2.5 fold higher than that of the cells transfected with the pIRES plasmid. The proliferation of the prostate smooth muscle cells was significantly inhibited by the TFPI gene 4 days post gene transfection. Conclusion TFPI might have a regulatory effect on the proliferation of the prostatic smooth muscle cells, further investigation is needed to explore the underlying mechanisms.