重组截断型潜伏相关肽通过抑制 TGF-β/Smad 通路减轻肝纤维化的体外和体内研究。
Recombinant truncated latency-associated peptide alleviates liver fibrosis in vitro and in vivo via inhibition of TGF-β/Smad pathway.
机构信息
Heilongjiang Province Key Laboratory of Anti-Fibrosis Biotherapy, Mudanjiang Medical University, No. 3, Tongxiang Street, Aimin District, Mudanjiang, 157011, Heilongjiang, China.
College of Life Sciences, Mudanjiang Medical University, Mudanjiang, 157011, Heilongjiang, China.
出版信息
Mol Med. 2022 Jul 16;28(1):80. doi: 10.1186/s10020-022-00508-2.
BACKGROUND
Liver fibrosis is a progressive liver injury response. Transforming growth factor β1 (TGF-β1) is oversecreted during liver fibrosis and promotes the development of liver fibrosis. Therapeutic approaches targeting TGF-β1 and its downstream pathways are essential to inhibit liver fibrosis. The N-terminal latency-associated peptide (LAP) blocks the binding of TGF-β1 to its receptor. Removal of LAP is critical for the activation of TGF-β1. Therefore, inhibition of TGF-β1 and its downstream pathways by LAP may be a potential approach to affect liver fibrosis.
METHODS
Truncated LAP (tLAP) plasmids were constructed. Recombinant proteins were purified by Ni affinity chromatography. The effects of LAP and tLAP on liver fibrosis were investigated in TGF-β1-induced HSC-T6 cells, AML12 cells and CCl-induced liver fibrosis mice by real time cellular analysis (RTCA), western blot, real-time quantitative PCR (RT-qPCR), immunofluorescence and pathological staining.
RESULTS
LAP and tLAP could inhibit TGF-β1-induced AML12 cells inflammation, apoptosis and EMT, and could inhibit TGF-β1-induced HSC-T6 cells proliferation and fibrosis. LAP and tLAP could attenuate the pathological changes of liver fibrosis and inhibit the expression of fibrosis-related proteins and mRNAs in CCl-induced liver fibrosis mice.
CONCLUSION
LAP and tLAP could alleviate liver fibrosis in vitro and in vivo via inhibition of TGF-β/Smad pathway. TLAP has higher expression level and more effective anti-fibrosis activity compared to LAP. This study may provide new ideas for the treatment of liver fibrosis.
背景
肝纤维化是一种进行性肝损伤反应。转化生长因子 β1（TGF-β1）在肝纤维化时过度分泌，并促进肝纤维化的发展。针对 TGF-β1 及其下游途径的治疗方法对于抑制肝纤维化至关重要。N 端潜伏相关肽（LAP）阻止 TGF-β1 与其受体结合。去除 LAP 对于 TGF-β1 的激活至关重要。因此，通过 LAP 抑制 TGF-β1 及其下游途径可能是影响肝纤维化的一种潜在方法。
方法
构建截断 LAP（tLAP）质粒。通过 Ni 亲和力层析法纯化重组蛋白。通过实时细胞分析（RTCA）、western blot、实时定量 PCR（RT-qPCR）、免疫荧光和病理染色，研究 LAP 和 tLAP 对 TGF-β1 诱导的 HSC-T6 细胞、AML12 细胞和 CCl 诱导的肝纤维化小鼠的作用。
结果
LAP 和 tLAP 可抑制 TGF-β1 诱导的 AML12 细胞炎症、凋亡和 EMT，可抑制 TGF-β1 诱导的 HSC-T6 细胞增殖和纤维化。LAP 和 tLAP 可减轻 CCl 诱导的肝纤维化小鼠的肝纤维化病理变化，并抑制纤维化相关蛋白和 mRNAs 的表达。
结论
LAP 和 tLAP 通过抑制 TGF-β/Smad 途径在体外和体内减轻肝纤维化。与 LAP 相比，tLAP 具有更高的表达水平和更有效的抗纤维化活性。本研究可能为肝纤维化的治疗提供新的思路。
Zotero 插件