CAJ | 학술논문

目的:探讨低分子量肝素对体外培养人早孕绒毛滋养层细胞MMP-2、TIMP-2表达及侵袭力的影响. 方法:将经胰蛋白酶/DNA酶Ⅰ联合消化,通过Percoll 细胞分离液纯化得到的绒毛滋养层细胞进行体外培养.采用不同浓度低分子量肝素干预培养24 h后,ELISA法测定细胞上清液中MMP-2、TIMP-2的浓度;采用Transwell小室观察滋养层细胞的侵袭能力.结果:不同浓度的低分子量肝素(1.0×10~2 IU/L,1.0×10~3 IU/L,1.0×10~4 IU/L)干预人早孕绒毛滋养层细胞后,与对照组相比,MMP-2的表达上调,滋养层细胞侵袭力增强,在1.0×10~3 IU/L时MMP-2表达最高,滋养层细胞侵袭力最强(P<0.05).TIMP-2的表达随着低分子量肝素浓度的增加而逐渐下降,与对照组比较,在1.0×10~3 IU/L、1.0×10~4 IU/L组明显降低(P<0.05),但1.0×10~3IU/L组与1.0×10~4 IU/L组之间TIMP-2的表达无差异(P>0.05). 结论:低分子量肝素可能直接通过影响绒毛滋养层细胞 MMP-2、TIMP-2的表达进而影响绒毛滋养层细胞的侵袭能力.
목적:탐토저분자량간소대체외배양인조잉융모자양층세포MMP-2、TIMP-2표체급침습력적영향. 방법:장경이단백매/DNA매Ⅰ연합소화,통과Percoll 세포분리액순화득도적융모자양층세포진행체외배양.채용불동농도저분자량간소간예배양24 h후,ELISA법측정세포상청액중MMP-2、TIMP-2적농도;채용Transwell소실관찰자양층세포적침습능력.결과:불동농도적저분자량간소(1.0×10~2 IU/L,1.0×10~3 IU/L,1.0×10~4 IU/L)간예인조잉융모자양층세포후,여대조조상비,MMP-2적표체상조,자양층세포침습력증강,재1.0×10~3 IU/L시MMP-2표체최고,자양층세포침습력최강(P<0.05).TIMP-2적표체수착저분자량간소농도적증가이축점하강,여대조조비교,재1.0×10~3 IU/L、1.0×10~4 IU/L조명현강저(P<0.05),단1.0×10~3IU/L조여1.0×10~4 IU/L조지간TIMP-2적표체무차이(P>0.05). 결론:저분자량간소가능직접통과영향융모자양층세포 MMP-2、TIMP-2적표체진이영향융모자양층세포적침습능력.
AIM: To investigate the hypothesis that low-molecular weight heparin (LMWH) regulates in vitro cytotrophoblast invasiveness and production of metalloproteinases-2 (MMP-2), tissue inhibitor of metalloproteinas-2 (TIMP-2). METHODS: Chorionic villi tissue of normal 6-8 weeks pregnancy was obtained. Trophoblastic cells were collected by trypsin-collagenase digestion and Percoll gradient centrifugation. The cytotrophoblastic cells were cultured for 24 h and divided into 4 groups according to the concentrations (1.0×10~2 IU/L, 1.0×10~3 IU/L or 1.0×10~4 IU/L) of LMWH adding into the medium. The contents of MMP-2 and TIMP-2 in cell culture supernatants were measured by the method of ELISA. Cytotrophoblast invasiveness was determined by Transwell chamber assay. RESULTS: With the increasing concentrations of LMWH, the invasion activity of cytotrophoblastic cells and MMP-2 secretion were increased. At concentration of 1.0×10~3IU/L, LMWH greatly enhanced cytotrophoblast invasiveness and the expression of MMP-2 (P<0.05). The levels of TIMP-2 were decreased after intervention with LMWH. At concentration of 1.0×10~3IU/L or 1.0×10~4 IU/L, LMWH induced a significant decrease in TIMP-2 expression. No significant difference between group 1×10~3IU/L and group 1.0×10~4 IU/L was observed (P>0.05). CONCLUSION: LMWH might regulate cytotrophoblast invasiveness in vitro by influencing the expression of MMP-2 and TIMP-2 in cytotrophoblastic cells.