HCV感染实验探讨人胎盘滋养层细胞分子生物性及侵袭力 FREE

Effect of HCV persistent infection on molecular biological characteristics and invasiveness of human placental trophoblast FREE

摘要:

目的 探讨持续感染丙型肝炎病毒（HCV）对体外培养人胎盘合体滋养层细胞分子生物学性状及其金属蛋白酶（MMP）2、MMP9合成及分泌的影响。方法 分别采用Matrigel人工重建基底膜侵袭实验、MTT法细胞黏附实验、细胞移动实验，研究HCV RNA阳性患者血清感染的体外培养人胎盘滋养层细胞(感染组)侵袭能力以及侵袭相关的黏附、移动能力的改变；检测培养上清中人绒毛膜促性腺激素（HCG）浓度以评估感染对细胞激素合成、分泌能力的影响。用酶联免疫吸附试验（ELISA）检测细胞培养上清MMP2和MMP9水平，分析感染组和对照组(健康体检者血清培养)数据间的差异，并用明胶酶谱进一步验证ELISA结果。应用逆转录聚合酶链反应（RTPCR）法分析感染对细胞proMMP2和proMMP9 mRNA表达的影响。 结果 感染组细胞侵袭、黏附、移动以及激素合成、分泌能力较对照组细胞均显著下降(P＜0.05)。感染组细胞分泌MMP2和MMP9能力明显下降（与对照组比较，MMP2：t=4.186，P＜0.05； MMP9：t=2.325，P＜0.05）；RTPCR结果显示感染组细胞proMMP2和proMMP9 mRNA表达弱于对照组，但差异无显著性（proMMP2：t=1.196，P＞0.05；proMMP9：t=1.417，P＞0.05）。
结论 持续HCV感染的体外培养人胎盘合体滋养层细胞MMP2和MMP9合成、分泌能力下降。持续感染HCV可抑制体外培养人胎盘合体滋养层细胞包括侵袭能力和激素合成、分泌能力在内的多种生物学功能。

Abstract:

Objective To explore  hepatitis C virus (HCV) persistent infection on biological characteristics, matrix metalloproteinase2 (MMP2) and MMP9 synthesis and secretion of cultured human placental syncytiotrophoblast(HPS).Methods The artificial reconstruction basement membrane Matrigel invasion assay, MTT  cell adhesion and cell motility experiments were used to study the change in invasion, invasionrelated adhesion and mobility of HPS(infected group) infected by serum of HCV RNA positive patients in vitro; concentrations of  human chorionic gonadotropin (HCG) cultured in supernatant was detected  to assess the impact of infection on cell  hormone synthesis and secretion. MMP2 and MMP9 levels in  cell culture supernatant were detected by enzyme linked immunosorbent assay(ELISA), the differences between  data of infected  and control group(serum culture of healthy persons)was compared, the results of ELISA was further validated by gelatin zymography, impact of infection on cell proMMP2 and proMMP9 mRNA expression was  analyzed by  RTPCR.Results The cell invasion, adhesion, motility as well as the ability of synthesis and secretion of hormone in infected group decreased  significantly  than those of  control group  (P<0.05). Secretion  of MMP2 and MMP9 in infected group decreased obviously compared with control group (MMP2: t=4.186,P<0.05; MMP9: t=2.325,P<0.05); RTPCR results showed that expression of  proMMP2 and proMMP9 mRNA in infected group was lower than  control group, but the difference was not significant(proMMP2: t=1.196, P>0.05; proMMP9: t=1.417, P>0.05). 
Conclusion The capacity  of HCV persistent infected HPS to synthesize and secrete MMP2 and MMP9 reduced. Persistent infection with HCV can inhibit  a variety of biological functions, including invasiveness and hormone synthesis and secretion of HPS cultured in vitro.

［Key words］hepatitis C virus；placenta； syncytiotrophoblast; molecular biology; MMP2;MMP9; in vitro test

［Chin Infect Control，2010，9（2）：76-80］