Effect of stretching on inflammation in a subcutaneous carrageenan mouse model analyzed at single-cell resolution

Understanding the factors that influence the biological response to inflammation is crucial, due to its involvement in physiological and pathological processes, including tissue repair/healing, cancer, infections, and autoimmune diseases. We have previously demonstrated that in vivo stretching can reduce inflammation and increase local pro-resolving lipid mediators in rats, suggesting a direct mechanical effect on inflammation resolution. Here, we aimed to explore further the effects of stretching at the cellular/molecular level in a mouse subcutaneous carrageenan-inflammation model. Stretching for 10 minutes twice a day reduced inflammation, increased the production of pro-resolving mediator pathway intermediate 17-HDHA at 48h post carrageenan injection, and decreased both pro-resolving and pro-inflammatory mediators (e.g., PGE2 and PGD2) at 96h. ScRNAseq analysis of inflammatory lesions at 96h showed that stretching increased the expression of both pro-inflammatory (Nos2) and pro-reso..., All ultrasound data acquisition and measurements were performed by investigators blinded to intervention condition. Ultrasound images of the back were acquired under isoflurane anesthesia. A high-frequency ultrasound scanner (Vevo 2100, Fujifilm VisualSonics, Toronto, Canada) in B mode with a 21 MHz transducer (MS 250) was used for optimal spatial resolution. A conductive gel was centrifuged for 5 minutes to remove air bubbles and spread over the skin. The transducer was stabilized with a clamp and mounted into an articulated arm to control the distance and the angle between the transducer and the skin surface. the transducer was oriented transversal or sagittal perpendicular to the skin of the back and centered on the lesion area. Total lesion area was calculated by averaging the lesion area measured at transversal and sagittal positions. Flow cytometry Inflammatory lesions were excised and minced in 5% FBS-DMEM, using a scalpel, then the suspension was filtered through a 70mm filter...., , # README \#**For the Ultrasound area:** \#Berrueta et al_Area measured by US (ultrasound) carrageenan inflammation 24-96h. The data is organized as follow: There are 6 columns, column A describe the code assigned to each individual animal. Column B represent treatment: 0=No stretch, 1= stretch. Column C refers to the time for the treatment: from 24h to 96h. Column D refers to the batch or group of samples. Column E correspond to the weight of the tissue at the time of euthanasia in mg. Column F represent the ultrasound area of the inflammatory lesion per each sample. \#**For the flow cytometry data** **Neutrophils and macrophage subpopulation**. Analysis was performed using Flow cytometry. The data set is organized as follow: \#**Berrueta et al_neutrophils dynamics**: There are 9 columns, column A indicates the code assigned to each individual animal. Column B represent treatment: 0=No stretch, 1= stretch. Column C refers to the time for the treatment: from 24h to 96h. Column D refe...阐明影响炎症生物学应答的关键因素具有重要意义，因为炎症广泛参与诸多生理与病理过程，涵盖组织修复、癌症、感染以及自身免疫性疾病等。此前我们的研究已证实，在体拉伸可降低大鼠体内炎症水平，并提升局部促消退脂质介质的表达，提示机械刺激可直接调控炎症消退过程。本研究旨在进一步探究拉伸在细胞/分子层面的调控效应，采用小鼠皮下角叉菜胶炎症模型开展实验。每日两次、每次10分钟的拉伸干预可减轻炎症反应：在角叉菜胶注射后48小时，可提升促消退介质通路中间产物17-HDHA的生成量；而在96小时时，可同时降低促消退与促炎介质（如PGE2、PGD2）的水平。对96小时炎症病灶的单细胞RNA测序（single-cell RNA sequencing, scRNAseq）分析显示，拉伸可同时上调促炎基因（Nos2）与促消退相关基因的表达…… 所有超声数据的采集与测量均由对干预分组不知情的实验人员完成。背部超声成像在异氟烷麻醉状态下进行。高频超声扫描仪（Vevo 2100，富士胶片VisualSonics公司，加拿大多伦多）搭配21 MHz换能器（MS 250）并启用B型模式，以获得最优空间分辨率。使用前将导电凝胶离心5分钟以去除气泡，随后涂抹于实验动物皮肤表面。通过夹具固定换能器，并将其安装于机械臂上，以精准控制换能器与皮肤表面的距离及夹角。将换能器以横断面或矢状面方向垂直于背部皮肤，并对准病灶区域中心。通过对横断面与矢状面测量得到的病灶面积取平均值，计算得到总病灶面积。 流式细胞术 使用手术刀将炎症病灶切除并切碎于含5%胎牛血清的DMEM培养基（5% FBS-DMEM）中，随后将细胞悬液通过70mm滤膜过滤…… # README # 超声病灶面积数据： # Berrueta等_超声测量的角叉菜胶炎症模型24-96小时病灶面积 数据集组织格式如下：共包含6列，A列为每只实验动物的编号；B列为干预分组：0=未拉伸，1=拉伸；C列为干预时间点：覆盖24h至96h；D列为样本批次/组别；E列为安乐死时采集的组织重量（单位：mg）；F列为每个样本的炎症病灶超声测量面积。 # 流式细胞术数据 **中性粒细胞与巨噬细胞亚群** 采用流式细胞术完成数据分析。数据集组织格式如下： # Berrueta等_中性粒细胞动态变化：共包含9列，A列为每只实验动物的编号；B列为干预分组：0=未拉伸，1=拉伸；C列为干预时间点：覆盖24h至96h；D列为……