Early activated hepatic stellate cell-derived molecules reverse acute hepatic injury
Early activated hepatic stellate cell-derived molecules reverse acute hepatic injury
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DOI：
10.3748/wjg.v21.i14.4184
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发表时间：
2015-04-14
影响因子：
4.3
通讯作者：
Qin, Xin-Yu
中科院分区：
文献类型：
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作者：
Chang, Wen-Ju;Song, Lu-Jun;Qin, Xin-Yu
目的：目的：探讨不同活化阶段的肝星状细胞（hepatic stellate cells，HSC）在醋氨酚（acetaminophen，APAP）诱导的急性肝损伤（acute liver injury，ALI）中的作用。用免疫荧光和透射电镜观察原始HSCs [HSCs（5d）]和永生化HSCs [HSCs（p3）]的形态学变化。HSC衍生分子、细胞裂解物和HSC条件培养基（HSC-CM）的保护作用通过存活和组织病理学分析在体内测试。通过测量血清中的转氨酶水平和在光学显微镜下对组织切片进行组织学检查来确定肝损伤。此外，为了确定所观察到的启动HSC的保护作用的分子介质，我们使用高密度蛋白阵列检测HSC-CM。HSC（5d）呈星形外观，细胞间表达水平不均匀。然而，HSC（p3）进化成肌纤维母细胞样细胞，没有脂滴，并表达均匀和更高水平的α-SMA。HSC-CM（5d），而不是HSC-CM（p3），提供了一个显着的生存优势，并显示出显着减少肝细胞坏死和全小叶白细胞浸润暴露于APAP的小鼠。然而，这种保护作用在较高的细胞质量下被废除，表明有效性的治疗窗口。此外，蛋白质芯片筛选显示HSC-CM（5d）由多种与炎症抑制和治疗活性相关的趋化因子和生长因子组成。当与HSC-CM（p3）相比时，更高水平的单核细胞趋化蛋白-1、巨噬细胞炎性蛋白-1、HSC-CM（5d）中肝细胞生长因子、白细胞介素-10、基质金属蛋白酶-2的表达明显升高，而干细胞因子和Fas-配体的表达明显降低。结论：HSC-CM（5d）中起始HSC-CM和永生化HSC-CM在形态和蛋白表达上存在差异，为起始HSC-CM治疗ALI的潜在医学价值提供了实验依据。
AIM: To test whether hepatic stellate cells (HSCs) at different activation stages play different roles in acetaminophen (APAP)-induced acute liver injury (ALI).METHODS: HSCs were isolated from mouse liver and cultured in vitro. Morphological changes of initiation HSCs [HSCs (5d)] and perpetuation HSCs [HSCs (p3)] were observed by immunofluorescence and transmission electron microscopy. The protective effects of HSC-derived molecules, cell lysates and HSC-conditioned medium (HSC-CM) were tested in vivo by survival and histopathological analyses. Liver injury was determined by measuring aminotransferase levels in the serum and by histologic examination of tissue sections under a light microscope. Additionally, to determine the molecular mediators of the observed protective effects of initiation HSCs, we examined HSC-CM using a high-density protein array.RESULTS: HSCs (5d) and HSCs (p3) had different morphological and phenotypic traits. HSCs (5d) presented a star-shaped appearance with expressing a-SMA at non-uniform levels between cells. However, HSCs (p3) evolved into myofibroblast-like cells without lipid droplets and expressed a uniform and higher level of a-SMA. HSC-CM (5d), but not HSC-CM (p3), provided a significant survival benefit and showed a dramatic reduction of hepatocellular necrosis and panlobular leukocyte infiltrates in mice exposed to APAP. However, this protective effect was abrogated at higher cell masses, indicating a therapeutic window of effectiveness. Furthermore, the protein array screen revealed that HSC-CM (5d) was composed of many chemokines and growth factors that correlated with inflammatory inhibition and therapeutic activity. When compared with HSC-CM (p3), higher levels of monocyte chemoattractant protein-1, macrophage inflammatory protein-1., hepatocyte growth factor, interleukin-10, and matrix metalloproteinase-2, but lower levels of stem cell factor and Fas-Ligand were observed in HSC-CM (5d).CONCLUSION: These data indicated that initiation HSCs and perpetuation HSCs were different in morphology and protein expression, and provided the first experimental evidence of the potential medical value of initiation HSC-derived molecules in the treatment of ALI.