CAJ | 학술논문

目的探讨子宫内膜异位症(内异症)患者腹腔液中正常T淋巴细胞表达和分泌调节活化因子(RANTES)分泌水平的变化及其对单核巨噬细胞趋化的影响.方法建立内异症异位病灶主要细胞--子宫内膜基质细胞(ESC)、腹膜间皮细胞(HPMC)及单核巨噬细胞系U937不同组合的直接接触(E-H-U、E-H、E-U、H-U)及间接接触共培养体系(U/E-H、H/E-U、E/H-U),然后收集共培养上清液,用酶联免疫吸附试验(ELISA)检测RANTES分泌水平的变化.利用趋化实验观察ESC、HPMC及ESC-HPMC共培养对单核巨噬细胞的趋化作用.结果 ESC、HPMC、U937单独培养时,RANTES的分泌水平分别为(5.0±0.5)、(4.0±0.3)、(254±40)ng/L;与单独培养比较,共培养明显促进RANTES的分泌,其中E-H-U培养为(2250±96) ng/L,E-U培养为(243±192) ng/L,H-U培养为(1251±73) ng/L,E-H培养为(50±40) ng/L,3种细胞直接共培养较两种细胞直接共培养时,RANTES分泌水平明显升高,差异有统计学意义(P＜0.01);间接共培养时,RANTES的分泌水平分别为(1519±96) ng/L(E/H-U培养)、(912±93) ng/L(U/E-H培养)、(1201±93)ng/L(H/E-U培养).ESC、HPMC及ESC-HPMC共培养可促进单核巨噬细胞的趋化,迁移细胞数分别为(6.0±0.3)、(6.2±0.3)、(10.0±0.3)×103个.结论内异症病灶主要细胞间存在着复杂的相互作用机制,并可促进趋化因子RANTES的分泌,从而促进腹腔内单核巨噬细胞的趋化和募集.
목적 탐토자궁내막이위증(내이증)환자복강액중정상T림파세포표체화분비조절활화인자(RANTES)분비수평적변화급기대단핵거서세포추화적영향.방법 건립내이증이위병조주요세포--자궁내막기질세포(ESC)、복막간피세포(HPMC)급단핵거서세포계U937불동조합적직접접촉(E-H-U、E-H、E-U、H-U)급간접접촉공배양체계(U/E-H、H/E-U、E/H-U),연후수집공배양상청액,용매련면역흡부시험(ELISA)검측RANTES분비수평적변화.이용추화실험관찰ESC、HPMC급ESC-HPMC공배양대단핵거서세포적추화작용.결과 ESC、HPMC、U937단독배양시,RANTES적분비수평분별위(5.0±0.5)、(4.0±0.3)、(254±40)ng/L;여단독배양비교,공배양명현촉진RANTES적분비,기중E-H-U배양위(2250±96) ng/L,E-U배양위(243±192) ng/L,H-U배양위(1251±73) ng/L,E-H배양위(50±40) ng/L,3충세포직접공배양교량충세포직접공배양시,RANTES분비수평명현승고,차이유통계학의의(P＜0.01);간접공배양시,RANTES적분비수평분별위(1519±96) ng/L(E/H-U배양)、(912±93) ng/L(U/E-H배양)、(1201±93)ng/L(H/E-U배양).ESC、HPMC급ESC-HPMC공배양가촉진단핵거서세포적추화,천이세포수분별위(6.0±0.3)、(6.2±0.3)、(10.0±0.3)×103개.결론 내이증병조주요세포간존재착복잡적상호작용궤제,병가촉진추화인자RANTES적분비,종이촉진복강내단핵거서세포적추화화모집.
Objective To explore the secretion of chemokine regulated upon activation normal T cell expressed and secreted(RANTES)influenced by the complex microenvironment in the peritoneal cavity of women with endometriosis and investigate chemotaxis of RANTES on the peritoneal monocytes.Methods The contact and non-contact co-culture systems including three target cells of ectopic tissue were established.The three target cells were endometrial stromal cells(ESC),human peritoneal mesothelial cells(HPMC)and monocytes.After collection of the supernatant of co-culture systems,the levels of RANTES were detected by enzyme linked immunosorbent assay(EUSA).Migration of U937 cell,a monocyte line,was detected by chemotaxis assay.Results ESC,HPMC,and U937 cultured alone secreted slight RANTES,(5.0±0.5),(4.0±0.3),and (254±40)ng/L. Compared with the culture of the target cell alone,the levels of RANTES in each co-culture system increased significantly,with the highest level in the contact culture system of E-H-U(2250±96)ng/L. RANTES secretion of non-contact co-culture of three cells were higher than contact co-culture of two cells(P＜0.01):U/E-H(912±93) vs E-H(50±40)ng/L,H/E-U(1201±93) vs E-U(243±192)ng/L,and E/H-U(1519±96) vs H-U(1251±73)ng/L. ESC,HPMC,and ESC-HPMC co-culture improved significantly migration of U937 cells [ number of cell migration respectively(6.0±0.3),(6.2±0.3),(10.0±0.3)×103,P＜0.01],which could be inhibited efficiently by anti-RANTES neutralizing antibody.Condusion The target cells in the peritoneal cavity of patients with endometriosis promote the secretion of RANTES in autocrine and paracrine manners and migration of monocytes.