脓毒症相关急性肾损伤后，脂肪源性干细胞源性细胞外囊泡通过介导ADAM17/MerTK介导小管上皮细胞凋亡影响巨噬细胞efferocysis的机制

IF 6.4 2区医学 Q1 MEDICAL LABORATORY TECHNOLOGY

Translational Research Pub Date : 2025-05-20 DOI:10.1016/j.trsl.2025.05.002

Zhixiang Bian, Xiangxiang Wang, Xiaoxuan Su, Ming Yang, Rui Zhu, Shunjie Chen

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Mouse serum urea and creatinine, and KIM-1, efferocytosis- and apoptosis-related protein, inflammatory factor, cytokine, and soluble MerTK (sMerTK) levels were determined using colorimetric assay, immunohistochemistry, Western blot, and ELISA. Renal tubular injury, TEC apoptosis, macrophage efferocytosis, and M1/M2 polarization levels were assessed via HE staining, TUNEL staining, immunofluorescence, and flow cytometry, respectively. <em>In vivo</em> validation experiments were conducted.</div></div><div><h3>Results</h3><div>S-AKI mice displayed elevated levels of serum urea, creatinine, KIM-1, pro-inflammatory factors, pro-apoptotic proteins and ADAM17 protein, decreased anti-apoptotic protein and MerTK protein levels, and diminished M2 polarization. ADSC-EVs down-regulated ADAM17 and sMerTK, and increased cell membrane MerTK, macrophage recognition of apoptotic cells and efferocytosis, and M2 polarization in renal tissues of S-AKI mice and LPS-induced mouse renal macrophages, indicating that ADSC-EVs regulated ADAM17/MerTK-mediated macrophage efferocytosis and promoted M2 polarization. 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引用次数: 0

摘要

目的：探讨脂肪源性干细胞源性细胞外囊泡（ADSC-EVs）通过调节ADAM17/ mertk介导的巨噬细胞efferocytosis改善脓毒症后急性肾损伤（S-AKI）小管上皮细胞（TEC）凋亡的分子机制。方法：采用结扎法和穿刺法建立小鼠S-AKI模型，并静脉注射adsc - ev。小鼠肾巨噬细胞用LPS培养，用ev培养，同时转染oe-ADAM17或si-MerTK，然后与Jurkat细胞孵育。采用比色法、免疫组织化学、Western blot和ELISA检测小鼠血清尿素和肌酐、KIM-1、efferocytosis和凋亡相关蛋白、炎症因子、细胞因子和可溶性MerTK （sMerTK）水平。分别通过HE染色、TUNEL染色、免疫荧光和流式细胞术评估肾小管损伤、TEC凋亡、巨噬细胞efferocyte和M1/M2极化水平。进行了体内验证实验。结果：S-AKI小鼠血清尿素、肌酐、KIM-1、促炎因子、促凋亡蛋白和ADAM17蛋白水平升高，抗凋亡蛋白和MerTK蛋白水平降低，M2极化减弱。adsc - ev下调ADAM17和sMerTK，增加S-AKI小鼠肾组织和lps诱导的小鼠肾巨噬细胞细胞膜MerTK、巨噬细胞对凋亡细胞和efferocysis的识别以及M2极化，表明adsc - ev调节ADAM17/MerTK介导的巨噬细胞efferocysis，促进M2极化。MerTK沉默部分逆转了adsc - ev调节的lps诱导的小鼠肾巨噬细胞efferocysis和M2极化。在体内，ADAM17上调部分避免了adsc - ev调控的s- aki后小鼠肾组织TEC凋亡。结论：adsc - ev通过调节ADAM17下调巨噬细胞sMerTK水平，上调巨噬细胞膜MerTK蛋白水平，促进巨噬细胞effocysis，改善s - aki TEC后细胞凋亡和炎症。

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Mechanism of adipose-derived stem cell-derived extracellular vesicles affecting macrophage efferocytosis by mediating ADAM17/MerTK in the apoptosis of tubular epithelial cells after sepsis-associated acute kidney injury

Objective

This study explored the molecular mechanism of adipose-derived stem cell-derived extracellular vesicles (ADSC-EVs) improving post-sepsis-associated acute kidney injury (S-AKI) tubular epithelial cell (TEC) apoptosis by modulating ADAM17/MerTK-mediated macrophage efferocytosis.

Methods

The S-AKI mouse model was established by caecal ligation and puncture and intravenously injected with ADSC-EVs. Mouse kidney macrophages were cultured with LPS, cultured with EVs while transfecting with oe-ADAM17 or si-MerTK, then incubated with Jurkat cells. Mouse serum urea and creatinine, and KIM-1, efferocytosis- and apoptosis-related protein, inflammatory factor, cytokine, and soluble MerTK (sMerTK) levels were determined using colorimetric assay, immunohistochemistry, Western blot, and ELISA. Renal tubular injury, TEC apoptosis, macrophage efferocytosis, and M1/M2 polarization levels were assessed via HE staining, TUNEL staining, immunofluorescence, and flow cytometry, respectively. In vivo validation experiments were conducted.

Results

S-AKI mice displayed elevated levels of serum urea, creatinine, KIM-1, pro-inflammatory factors, pro-apoptotic proteins and ADAM17 protein, decreased anti-apoptotic protein and MerTK protein levels, and diminished M2 polarization. ADSC-EVs down-regulated ADAM17 and sMerTK, and increased cell membrane MerTK, macrophage recognition of apoptotic cells and efferocytosis, and M2 polarization in renal tissues of S-AKI mice and LPS-induced mouse renal macrophages, indicating that ADSC-EVs regulated ADAM17/MerTK-mediated macrophage efferocytosis and promoted M2 polarization. MerTK silencing partially reversed ADSC-EVs-regulated LPS-induced mouse renal macrophage efferocytosis and M2 polarization. In vivo, ADAM17 upregulation partly averted ADSC-EVs-regulated post-S-AKI TEC apoptosis in mouse renal tissues.

Conclusion

ADSC-EVs down-regulated sMerTK level and up-regulated macrophage membrane MerTK protein level by modulating ADAM17 to promote macrophage efferocytosis and ameliorate post-S-AKI TEC apoptosis and inflammation.

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来源期刊

Translational Research 医学-医学：内科

CiteScore

15.70

自引率

0.00%

发文量

195

审稿时长

14 days

期刊介绍： Translational Research (formerly The Journal of Laboratory and Clinical Medicine) delivers original investigations in the broad fields of laboratory, clinical, and public health research. Published monthly since 1915, it keeps readers up-to-date on significant biomedical research from all subspecialties of medicine.