Sanyal A J, Mirshahi F
Department of Internal Medicine, Medical College of Virginia campus of Virginia Commonwealth University, Richmond, VA 23298-0711, USA.
Hepatology. 1999 Mar;29(3):710-8. doi: 10.1002/hep.510290323.
The phenotype of the endothelial cells (ECs) in the pseudointima of transjugular intrahepatic portasystemic shunts (TIPS) and the mechanisms of pseudointima formation after TIPS were unknown. We hypothesized that TIPS were lined by hepatic sinusoidal ECs, which stimulated the migration of smooth muscle cells (SMCs) into the pseudointima and their proliferation. Studies were done with the following specific aims: (1) isolation of ECs from TIPS pseudointima and comparison of their phenotype with human cirrhotic sinusoidal and vascular ECs derived from hepatic and portal veins as well as aorta, and (2) testing of the effects of TIPS ECs on TIPS-derived SMC migration and proliferation. ECs were isolated from eight TIPS retrieved from liver explants by immunomagnetic separation using monodispersed magnetizable polystyrene beads (Dynabeads M-450) coated with Ulex Europeus 1. EC phenotypes were examined by transmission electron microscopy, factor VIII-related antigen, CD31, CD14, and CD34 expression, uptake of acetylated LDL and secretion of type IV collagen. The effects of EC-conditioned media on SMC migration and proliferation were tested in multiwell chemotaxis chambers and by cell counting, respectively. ECs were obtained from TIPS pseudointima with >95% purity. The phenotype of TIPS-derived ECs matched that of cirrhotic sinusoidal endothelium (both expressed CD14) and differed from that of vascular endothelium (CD14 negative, Weibel-Palade positive). Conditioned media from both stenosed (n = 3) and nonstenosed (n = 3) TIPS-derived endothelial cells produced a marked (>100%) P <.001 increase in migration as well as (up to 88%) P <.01 proliferation of SMCs from both stenosed (n = 3) as well as nonstenosed TIPS (n = 3). These data indicate that TIPS pseudointima are lined by hepatic sinusoidal endothelial cells, which stimulate pseudointima formation by increasing SMC migration and proliferation.
经颈静脉肝内门体分流术（TIPS）假内膜中内皮细胞（ECs）的表型以及TIPS术后假内膜形成的机制尚不清楚。我们推测TIPS内衬肝窦内皮细胞，其刺激平滑肌细胞（SMCs）迁移至假内膜并使其增殖。本研究旨在实现以下具体目标：（1）从TIPS假内膜中分离ECs，并将其表型与源自肝静脉、门静脉以及主动脉的人类肝硬化窦状内皮细胞和血管内皮细胞进行比较；（2）检测TIPS来源的ECs对TIPS来源的SMC迁移和增殖的影响。通过使用包被有欧洲荆豆凝集素1的单分散可磁化聚苯乙烯珠（Dynabeads M - 450）进行免疫磁分离，从肝脏外植体中获取的8个TIPS中分离出ECs。通过透射电子显微镜、因子VIII相关抗原、CD31、CD14和CD34表达、乙酰化低密度脂蛋白摄取以及IV型胶原分泌来检测EC表型。分别在多孔趋化室中通过细胞计数检测EC条件培养基对SMC迁移和增殖的影响。从TIPS假内膜中获得的ECs纯度>95%。TIPS来源的ECs表型与肝硬化窦状内皮细胞匹配（均表达CD14），与血管内皮细胞不同（CD14阴性，Weibel - Palade阳性）。来自狭窄（n = 3）和非狭窄（n = 3）TIPS来源的内皮细胞的条件培养基均使来自狭窄（n = 3）和非狭窄TIPS（n = 3）的SMC迁移显著增加（>100%，P <.001）以及增殖增加（高达88%，P <.01）。这些数据表明，TIPS假内膜由肝窦内皮细胞内衬，其通过增加SMC迁移和增殖来刺激假内膜形成。
