Rubicon promotes the M2 polarization of Kupffer cells via LC3-associated phagocytosis-mediated clearance to improve liver transplantation
Rubicon promotes the M2 polarization of Kupffer cells via LC3-associated phagocytosis-mediated clearance to improve liver transplantation
复制标题
Rubicon 通过 LC3 相关的吞噬作用介导的清除促进 Kupffer 细胞的 M2 极化，以改善肝移植
DOI：
10.1016/j.cellimm.2022.104556
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发表时间：
2022-06-11
影响因子：
4.3
通讯作者：
Gong, Jianping
中科院分区：
文献类型：
--
作者：
Chen, Yajun;He, Yiping;Gong, Jianping
背景：肝移植术后急性排斥反应（AR）与术后患者的生存密切相关。方法：将Rubicon慢病毒（LV）和Rubicon小干扰RNA （siRNA）转染到小鼠肝组织提取的KCs中。提取原代KCs，与酶酵素和凋亡T淋巴细胞共培养。通过Western blotting （WB）和q-PCR检测CD86、CD163、IL-10、tnf - α、tgf - β、JAK1、STAT6、AKT1、mTOR和过氧化物酶体增殖物激活受体γ (PPAR γ)的水平。流式细胞术检测CD86和CD163的水平。将mCherry-GFP-LC3腺病毒（AV）转染KCs。免疫荧光法检测LC3II的募集及吞噬体与溶酶体的融合情况。将Rubicon腺相关病毒（AAV）经门静脉转染小鼠肝组织，建立肝移植后免疫耐受（IT）和AR模型。HE染色检测肝组织病理变化。TUNEL染色检测凋亡细胞。免疫组化染色检测KCs的极化状态。结果：rubicon介导的lc3相关吞噬作用（LAP）促进KCs降解和清除凋亡T淋巴细胞的能力。凋亡T淋巴细胞降解的产物多不饱和脂肪酸（PUFAs）激活PPAR γ，进一步促进KCs的M2极化。Rubicon介导的降解增强有助于促进KCs的M2极化和支持IT的微环境。结论：rubicon介导的LAP通过pufa依赖性PPAR γ激活促进KCs的清除能力和M2极化，从而改善LT。
Background: Acute rejection (AR) after liver transplantation (LT) is closely related to the survival of patients after surgery. Enhancement of the ability of Kupffer cells (KCs) to eliminate apoptotic cells can effectively alleviate AR.Methods: Rubicon lentivirus (LV) and Rubicon small interfering RNA (siRNA) were transfected into KCs extracted from the liver tissue of mice. Primary KCs were extracted and cocultured with zymosan and apoptotic T lymphocytes. The levels of CD86, CD163, IL-10, TNF-alpha, TGF-beta, JAK1, STAT6, AKT1, mTOR and peroxisome proliferator-activated receptor-gamma (PPAR gamma) were assessed via Western blotting (WB) and q-PCR. The levels of CD86 and CD163 were assessed via flow cytometry. mCherry-GFP-LC3 adenovirus (AV) was transfected into KCs. The recruitment of LC3II and the fusion of phagosomes and lysosomes were detected using immunofluorescence. Rubicon adeno-associated virus (AAV) was transfected into the liver tissue of mice via the portal vein, and models of immune tolerance (IT) and AR following LT were established. Pathological changes in the liver tissue were detected using HE staining. Apoptotic cells were assessed via TUNEL staining. The polarization state of KCs was detected via immunohistochemical staining.Results: Rubicon-mediated LC3-associated phagocytosis (LAP) promotes the ability of KCs to degrade and clear apoptotic T lymphocytes. Polyunsaturated fatty acids (PUFAs), the product of apoptotic T lymphocyte degradation, activate PPAR gamma, which further promotes the M2 polarization of KCs. Enhanced degradation mediated by Rubicon contributes to promoting the M2 polarization of KCs and a microenvironment supportive of IT.Conclusions: Rubicon-mediated LAP promotes the clearance capability and M2 polarization of KCs via PUFAdependent PPAR gamma activation to improve LT.