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目的:在妊娠过程中,胎盘可能暴露于多种病原微生物,威胁胎儿正常生长发育。为探讨人胎盘绒毛组织是否表达AIM2炎性体成员基因以及人胎盘组织的AIM2炎性体的活化形式。方法:以THP-1细胞来源的RNA和蛋白作为阳性对照,分别应用RT-PCR和Western blot方法检测人早孕期胎盘绒毛组织中AIM2炎性体两个相关基因AIM2和ASC的表达。分离和体外培养人胎盘绒毛膜组织,并用不同浓度的poly(d A:d T)进行转染,处理24小时后,分别收集组织培养上清和蛋白裂解液,Western blot检测蛋白裂解液中caspase-1的活化,ELISA检测培养上清中IL-1β的分泌。结果:RT-PCR和Western blot结果均显示人早孕期胎盘绒毛组织组成性表达AIM2炎性体相关基因AIM2和ASC。同时,体外培养的人胎盘绒毛组织在转染5μg/m L poly(d A:d T)后,caspase-1剪切片段p10显著增多,培养上清中IL-1β分泌也显著增多(P<0.01)。结论:人胎盘绒毛组织存在功能性的AIM2炎性体,能够被胞内双链DNA活化。
Objective: During pregnancy, the placenta may be exposed to a variety of pathogenic microorganisms, threatening the normal growth and development of the fetus. To investigate whether human placental villi express the AIM2 inflammasome member genes and the activated form of human placental AIM2 inflammasome. METHODS: RNA and protein from THP-1 cells were used as positive control. The expression of AIM2 and ASC were detected by RT-PCR and Western blot respectively in human placenta during pregnancy. The human placenta chorionic tissues were isolated and cultured in vitro. The cells were transfected with poly (d A: d T) in different concentrations. Twenty-four hours later, the tissue culture supernatants and protein lysates were collected. Western blot was used to detect the expression of caspase- 1 activation, ELISA detection of culture supernatant IL-1β secretion. Results: The results of RT-PCR and Western blot showed that the placental villi in the early pregnancy had the constitutive expression of the AIM2 inflammasome related genes AIM2 and ASC. At the same time, p10 of caspase-1 cleavage increased significantly in cultured human placenta villi after transfection of 5μg / ml poly (d A: d T), and the secretion of IL-1β in culture supernatant also increased significantly (P < 0.01). Conclusion: AIM2 inflammasome exists in human placenta villi and can be activated by intracellular double stranded DNA.