Department of Pulmonary and Critical Care Medicine, Zhongshan Hospital; Institute for Clinical Science Shanghai Institute of Clinical Bioinformatics Shanghai Engineering Research for AI Technology for Cardiopulmonary Diseases Jinshan Hospital Centre for Tumor Diagnosis and Therapy, Fudan University Shanghai Medical College, Shanghai, China.
Cell Biol Toxicol. 2023 Apr;39(2):451-465. doi: 10.1007/s10565-021-09670-5. Epub 2022 Jan 3.
Telocytes (TCs) are experimentally evidenced as an alternative of cell therapies for organ tissue injury and repair. The aims of the present studies are to explore direct roles of TCs and the roles of TC-derived exosomes in support of experimental acute lung injury (ALI) in vivo or in vitro.
The roles of TCs in experimental ALI were firstly estimated. Phosphoinositide 3-kinase (PI3K) p110δ and α/δ/β isoform inhibitors were used in study dynamic alterations of bio-behaviors, and in expression of functional factors of TCs per se and TC-co-cultured airway epithelial cells during the activation with lipopolysaccharide (LPS). TC-driven exosomes were furthermore characterized for intercellular communication by which activated or non-activated TCs interacted with epithelia.
Our results showed that TCs mainly prevented from lung tissue edema and hemorrhage and decreased the levels of VEGF-A and MMP9 induced by LPS. Treatment with CAL101 (PI3K p110δ inhibitor) and LY294002 (PI3Kα/δ/β inhibitor) could inhibit TC movement and differentiation and increase the number of dead TCs. The expression of Mtor, Hif1α, Vegf-a, or Mmp9 mRNA increased in TCs challenged with LPS, while Mtor, Hif1α, and Vegf-a even more increased after adding CAL101 or Mtor after adding LY. The rate of epithelial cell proliferation was higher in co-culture of human bronchial epithelial (HBE) and TCs than that in HBE alone under conditions with or without LPS challenge or when cells were treated with LPS and CAL101 or LY294002. The levels of mTOR, HIF1α, or VEGF-A significantly increased in mono-cultured or co-cultured cells, challenged with LPS as compared with those with vehicle. LPS-pretreated TC-derived exosomes upregulated the expression of AKT, p-AKT, HIF1α, and VEGF-A protein of HBE.
The present study demonstrated that intraperitoneal administration of TCs ameliorated the severity of lung tissue edema accompanied by elevated expression of VEGF-A. TCs could nourish airway epithelial cells through nutrients produced from TCs, increasing epithelial cell proliferation, and differentiation as well as cell sensitivity to LPS challenge and PI3K p110δ and α/δ/β inhibitors, partially through exosomes released from TCs.
电浆细胞（telocytes，TCs）已被实验证明是器官组织损伤和修复的细胞治疗的一种替代方法。本研究的目的是探讨 TCs 的直接作用以及 TC 衍生的外泌体在支持体内或体外实验性急性肺损伤（ALI）中的作用。
首先评估 TCs 在实验性 ALI 中的作用。在 LPS 刺激下，使用磷酸肌醇 3-激酶（PI3K）p110δ 和 α/δ/β 同工型抑制剂研究 TCs 本身及其与 TC 共培养的气道上皮细胞的生物行为和功能因子表达的动态变化。此外，通过激活或非激活 TCs 与上皮细胞的相互作用，对 TC 驱动的外泌体进行细胞间通讯的特征描述。
我们的结果表明，TCs 主要防止肺组织水肿和出血，并降低 LPS 诱导的 VEGF-A 和 MMP9 水平。用 CAL101（PI3K p110δ 抑制剂）和 LY294002（PI3Kα/δ/β 抑制剂）处理可抑制 TC 运动和分化，并增加 TC 死亡数量。LPS 刺激后 TCs 中 Mtor、Hif1α、Vegf-a 或 Mmp9 mRNA 的表达增加，而添加 CAL101 或 LY 后 Mtor 甚至更多增加。在 LPS 刺激或加入 LPS 和 CAL101 或 LY294002 时，与单独培养的 HBE 相比，HBE 与 TCs 共培养时上皮细胞的增殖率更高。与载体相比，用 LPS 预处理的 TC 衍生的外泌体上调了 LPS 刺激的单核或共培养细胞中 AKT、p-AKT、HIF1α 和 VEGF-A 蛋白的表达。
本研究表明，腹腔内给予 TCs 可改善肺组织水肿的严重程度，同时伴有 VEGF-A 表达升高。TCs 可以通过自身产生的营养物质滋养气道上皮细胞，增加上皮细胞的增殖和分化，以及对 LPS 刺激和 PI3K p110δ 和 α/δ/β 抑制剂的敏感性，部分通过 TCs 释放的外泌体。
