ADAMTS L1通过白细胞介素-6介导羊膜细胞凋亡对早产的潜在影响

ADAMTS L1 may increase the preterm birth by regulating apoptosis of amniotic membrane cell through interleukin-6

目的研究含凝血酶敏感蛋白基序的解聚蛋白样金属蛋白酶L1(a disintegrin-like and metalloproteinase with thrombospondin L1,ADAMTS L1)是否通过参与炎症调控过程影响羊膜细胞凋亡,从而成为早产发病的潜在影响因子。方法收集足月产、自发性早产的胎盘组织20例,将其分为足月分娩组和自发性早产组,每组10例。利用实时荧光定量PCR及蛋白质印迹法(Western blot技术)检测其中ADAMTS L1和白细胞介素-6(interleukin-6,IL-6)的mRNA及蛋白水平;体外利用PMA活化THP-1细胞系模拟炎症环境,并通过慢病毒敲降人羊膜细胞系WISH中ADAMTS L1(敲低组),检测其中ADAMTS L1及IL-6的mRNA及蛋白水平,比较其与空白组的差别。结果相较于足月分娩组,自发性早产组ADAMTS L1及IL-6的mRNA水平分别约增加了11、15倍,蛋白水平分别约增加了9、12倍;同时,敲低ADAMTS L1减少WISH细胞凋亡,与空白组相比,敲低ADAMTS L1后,凋亡相关蛋白caspase3 mRNA水平降至空白组的0.35,蛋白水平降至空白组的0.30。结论本研究利用体外实验评估了炎症过程中巨噬细胞分泌的IL-6可以调节ADAMTS L1,进而引起自发性早产。但是,需要进一步的研究来证实我们的发现。

Objective To study whether a disintegrin-like and metalloproteinase with thrombospondin L1(ADAMTS L1)affects the apoptosis of amniotic membrane cells by participating in the process of interventional recombination,thereby becoming a potential factor of preterm birth.Methods First,we collected 20 placental tissues of full-term birth(FTB)and spontaneous premature birth(PTB),and divided them into FTB group(10 cases)and PTB group(10 cases).QRT-PCR and Western blot were used to detect the mRNA and protein levels of ADAMTS L1 and interleukin-6(IL-6).Then we activated THP-1 cell line by PMA to simulate the metabolic environment in vivo.ADAMTS L1 was knocked down by lentivirus in WISH cell line(knocked down group),mRNA and protein levels of ADAMTSL1 and IL-6 were detected,and the data of knocked down group were compared with control group.Results The expression of ADAMTS L1 in preterm tissues was significantly increased,according to the full term birth women,the mRNA level of ADAMTSL1 and IL-6 increased 11 and 15 times respectively,the expression of ADAMTSL1 and IL-6 protein increased 9 and 12 times respectively.Knockdown of ADAMTS L1 reduced WISH cell apoptosis.Compared with the control group,after knocking down ADAMTS L1,the mRNA level of apoptosis-related protein caspase3 dropped to 0.35 of the control group,and the protein level dropped to 0.3 of the control group.Conclusion This study used in vitro experiments to evaluate that ADAMTS L1 may promote apoptosis of human IL-6 secreted by macrophages during inflammation can regulate ADAMTS L1,which in turn can cause spontaneous preterm labor.However,further research is needed to confirm our findings.