人骨髓间充质干细胞来源的外泌体 circCDK13 通过 miR-17-5p/KAT2B 调控 MFGE8 的表达抑制肝纤维化。
hMSCs-derived exosome circCDK13 inhibits liver fibrosis by regulating the expression of MFGE8 through miR-17-5p/KAT2B.
机构信息
Department of Infectious Diseases, The Second Xiangya Hospital, Central South University, Changsha, Hunan, 410011, People's Republic of China.
出版信息
Cell Biol Toxicol. 2023 Apr;39(2):1-22. doi: 10.1007/s10565-022-09714-4. Epub 2022 Apr 29.
OBJECTIVE
To investigate the effects of human bone marrow mesenchymal stem cells (hMSCs)-derived exosome circCDK13 on liver fibrosis and its mechanism.
METHODS
Exosomes derived from hMSCs were extracted and identified by flow cytometry and osteogenic and adipogenic induction, and the expressions of marker proteins on the surface of exosomes were detected by western blot. Cell proliferation was measured by CCK8 assay, the expression of active markers of HSCs by immunofluorescence, and the expressions of fibrosis-related factors by western blot. A mouse model of liver fibrosis was established by intraperitoneal injection of thioacetamide (TAA). Fibrosis was detected by HE staining, Masson staining, and Sirius red staining. Western blot was utilized to test the expressions of PI3K/AKT and NF-κB pathway related proteins, dual-luciferase reporter assay and RIP assay to validate the binding between circCDK13 and miR-17-5p as well as between miR-17-5p and KAT2B, and ChIP to validate the effect of KAT2B on H3 acetylation and MFGE8 transcription.
RESULTS
hMSCs-derived exosomes inhibited liver fibrosis mainly through circCDK13. Dual-luciferase reporter assay and RIP assay demonstrated the binding between circCDK13 and miR-17-5p as well as between miR-17-5p and KAT2B. Further experimental results indicated that circCDK13 mediated liver fibrosis by regulating the miR-17-5p/KAT2B axis, and KAT2B promoted MFGE8 transcription by H3 acetylation. Exo-circCDK13 inhibited PI3K/AKT and NF-κB signaling pathways activation through regulating the miR-17-5p/KAT2B axis.
CONCLUSION
hMSCs-derived exosome circCDK13 inhibited liver fibrosis by regulating the expression of MFGE8 through miR-17-5p/KAT2B axis.
目的
探讨人骨髓间充质干细胞（hMSCs）衍生的外泌体 circCDK13 对肝纤维化的影响及其机制。
方法
通过流式细胞术和成骨及成脂诱导提取并鉴定 hMSCs 来源的外泌体，Western blot 检测外泌体表面标记蛋白的表达。通过 CCK8 检测细胞增殖，免疫荧光检测肝星状细胞（HSCs）活性标志物的表达，Western blot 检测纤维化相关因子的表达。采用腹腔注射硫代乙酰胺（TAA）建立肝纤维化小鼠模型。通过 HE 染色、Masson 染色和天狼猩红染色检测纤维化。Western blot 检测 PI3K/AKT 和 NF-κB 通路相关蛋白的表达，双荧光素酶报告实验和 RIP 实验验证 circCDK13 与 miR-17-5p 以及 miR-17-5p 与 KAT2B 的结合，ChIP 实验验证 KAT2B 对 H3 乙酰化和 MFGE8 转录的影响。
结果
hMSCs 来源的外泌体主要通过 circCDK13 抑制肝纤维化。双荧光素酶报告实验和 RIP 实验证实 circCDK13 与 miR-17-5p 以及 miR-17-5p 与 KAT2B 之间存在结合。进一步的实验结果表明，circCDK13 通过调节 miR-17-5p/KAT2B 轴来调节肝纤维化，KAT2B 通过 H3 乙酰化促进 MFGE8 转录。外泌体 circCDK13 通过调节 miR-17-5p/KAT2B 轴抑制 PI3K/AKT 和 NF-κB 信号通路的激活。
结论
hMSCs 衍生的外泌体 circCDK13 通过 miR-17-5p/KAT2B 轴调节 MFGE8 的表达来抑制肝纤维化。
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