[关键词]
[摘要]
目的：研究K-115对人Tenon囊成纤维细胞(HTFs)增殖和迁移的影响，并探讨其相关作用机制，为青光眼术后抗瘢痕治疗提供新思路。
方法：选取2018-09/2019-09于河北省人民医院行青光眼手术患者的Tenon囊组织，采用组织块法进行HTFs原代培养，应用转化生长因子-β1(TGF-β1)诱导HTFs模拟青光眼滤过性手术后的细胞活化模型，并采用K -115处理细胞。将细胞分为4组：对照组采用溶剂二甲基亚砜(DMSO)处理； TGF-β1组采用10μg/L TGF-β1处理24h； TGF-β1+5 K-115组采用5μmol/L K-115预处理2h后加入10μg/L TGF-β1处理24h； TGF-β1+10 K-115组采用10μmol/L K-115预处理2h后加入10μg/L TGF-β1处理24h。通过细胞增殖实验观察细胞的增殖能力； 划痕试验检测细胞的迁移能力； 透射电子显微镜观察细胞内自噬小体的形成； Hoechst 33342/PI染色观察细胞凋亡情况。
结果：细胞增殖实验结果提示K-115可以抑制TGF-β1诱导的HTFs增殖； 划痕试验提示K-115可以抑制TGF-β1诱导的HTFs迁移； 透射电子显微镜结果显示K-115可以增强TGF-β1诱导的HTFs自噬。Hoechst 33342/PI染色提示K-115并未诱导细胞凋亡。
结论：K-115可能是通过增加HTFs自噬而非诱导凋亡机制调控TGF-β1诱导的HTFs增殖及迁移能力。
[Key word]
[Abstract]
AIM:To investigate the influence of K-115 on the proliferation and migration of human Tenon's fibroblasts(HTFs)and to access the possible mechanism. Furthermore, to provide new ideas for anti-scar treatment after glaucoma surgery.
METHODS: The Tenon capsule tissues were collected from patients who underwent glaucoma surgery in Hebei General Hospital from September 2018 to September 2019. Primary culture of HTFs was performed by tissue block method. The transforming growth factor-β1(TGF-β1)was used to induce HTFs activation that can mimic glaucoma filtration surgery. The cells were treated with K-115 and divided into 4 groups: the control group was treated with dimethyl sulfoxide(DMSO); TGF-β1 group was treated with 10μg/L TGF-β1 for 24h; TGF-β1 +5 K-115 group was pretreated with 5μmol/L K-115 for 2h and then treated with 10μg/L TGF-β1 for 24h; TGF-β1+10 K-115 group was pretreated with 10μmol/L K-115 for 2h and then 10μg/L TGF-β1 was added for 24h. Cell proliferation was observed by cell proliferation experiment. The migration ability of cells was detected by scratch test. The formation of autophagosomes was observed by transmission electron microscopy. Apoptosis was visualized by Hoechst 33342/PI staining.
RESULTS: Cell proliferation experiment revealed that K-115 could inhibit the proliferation of HTFs induced by TGF-β1. Scratch test suggested that K-115 could inhibit the migration of HTFs induced by TGF-β1. Transmission electron microscope results showed that K-115 could enhance autophagy of HTFs induced by TGF-β1. Hoechst 33342/PI staining suggested that K-115 did not induce apoptosis.
CONCLUSIONS: K-115 may regulate the proliferation and migration of HTFs induced by TGF-β1 by increasing autophagy rather than inducing apoptosis.
[中图分类号]
[基金项目]
国家自然科学基金项目(No.81700835); 河北省省级科技计划项目(No.18277753D)
