Department of General Surgery, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, Jiangsu, China (mainland).
Med Sci Monit. 2019 Apr 17;25:2835-2844. doi: 10.12659/MSM.913481.
BACKGROUND This study aims to demonstrate the underlying correlation between the resolution of liver fibrosis induced by Gexia-Zhuyu decoction (GZD) treatment and myeloid cell-mediated angiogenesis. MATERIAL AND METHODS A liver fibrosis mouse model induced by carbon tetrachloride (CCl4) intervention was employed in this study. Dynamics of blood liver function parameters were followed. The liver pathology was detected by Sirius Red and Masson staining. Matrix metalloproteinase (MMP) 2/9, tissue inhibitors of metalloproteinase (TIMP)-1/2, and vascular endothelial growth factor (VEGF)-A expression levels were measured. Bone marrow chimera mice were generated by transfer of bone morrow cells from green fluorescent protein (GFP)-knockin mice into irradiated wild-type mice, and were used it to visualize the role of myeloid cells on the fibrosis resolution induced by GZD treatment. RESULTS The result of Sirius Red and Masson staining and the dynamics of blood liver function parameters showed that 5 weeks of GZD treatment attenuated the severity of liver fibrosis with continual CCl4 administration. GZD treatment promoted the expression of MMP2/9 and repressed the heightened level of TIMP-1/2 in the recovery phase. More notably, the increased VEGF-A and augmented endothelial progenitor cells were observed in the liver and blood in mice that received GZD, and contributed to the remodeling of hepatic vascular though the CXCL12/CXCR4 axis. Then, chimera mice with GFP-positive bone marrow cells were used to show angiogenesis driven by GZD-induced myeloid cell motivation. We found that GZD facilitated myeloid cells binding to the vascular CXCR4 and induced the resolution of fibrosis. CONCLUSIONS This study shows that activation of myeloid cells induced by GZD administration accelerates the functional angiogenesis, which benefits the resolution of CCl4-induced liver fibrosis.
本研究旨在展示肝纤维化的解决与骨髓细胞介导的血管生成之间的潜在关联。
本研究采用四氯化碳（CCl4）干预诱导的肝纤维化小鼠模型。监测血液肝功能参数的动态变化。通过天狼星红和 Masson 染色检测肝组织病理学变化。检测基质金属蛋白酶（MMP）2/9、组织抑制剂的金属蛋白酶（TIMP）-1/2 和血管内皮生长因子（VEGF）-A 的表达水平。通过将 GFP 敲入小鼠的骨髓细胞转移到照射的野生型小鼠中，生成骨髓嵌合小鼠，用于可视化骨髓细胞在 GZD 治疗诱导的纤维化消退中的作用。
天狼星红和 Masson 染色结果以及血液肝功能参数的动态变化表明，5 周的 GZD 治疗可减轻持续 CCl4 给药引起的肝纤维化严重程度。GZD 治疗在恢复阶段促进 MMP2/9 的表达，并抑制 TIMP-1/2 的升高水平。更值得注意的是，在接受 GZD 治疗的小鼠的肝脏和血液中观察到 VEGF-A 增加和内皮祖细胞增加，这有助于通过 CXCL12/CXCR4 轴重塑肝血管。然后，使用 GFP 阳性骨髓细胞的嵌合小鼠显示 GZD 诱导的骨髓细胞激活驱动的血管生成。我们发现，GZD 促进骨髓细胞与血管 CXCR4 的结合，并诱导纤维化的消退。
本研究表明，GZD 给药诱导的骨髓细胞的激活加速了功能性血管生成，有利于 CCl4 诱导的肝纤维化的解决。
