骨髓间充质干细胞条件培养基可改善大鼠糖尿病足溃疡。
Bone marrow-derived mesenchymal stem cell-conditioned medium ameliorates diabetic foot ulcers in rats.
机构信息
Department of Endocrinology, Air Force Hospital of Northern Theater Command of PLA, China.
Department of Endocrinology, Air Force Hospital of Northern Theater Command of PLA, China.
出版信息
Clinics (Sao Paulo). 2023 Mar 20;78:100181. doi: 10.1016/j.clinsp.2023.100181. eCollection 2023.
OBJECTIVES
This study aimed to explore the effects of bone marrow-derived Mesenchymal Stem Cell-Conditioned Medium (MSC-CM) treating diabetic foot ulcers in rats.
METHODS
Models of T2DM rats were induced by a high-fat diet and intraperitoneal injection of STZ in SD rats. Models of Diabetic Foot Ulcers (DFUs) were made by operation on hind limbs in diabetic rats. Rats were divided into four groups (n = 6 for each group), i.e., Normal Control group (NC), Diabetes Control group (DM-C), MSC-CM group and Mesenchymal Stem Cells group (MSCs). MSC-CM group was treated with an injection of conditioned medium derived from preconditioned rats' bone marrow MSCs around ulcers. MSCs group were treated with an injection of rats' bone marrow MSCs. The other two groups were treated with an injection of PBS. After the treatment, wound closure, re-epithelialization (thickness of the stratum granulosums of the skin, by H&E staining), cell proliferation (Ki67, by IHC), angiogenesis (CD31, by IFC), autophagy (LC3B, by IFC and WB; autolysosome, by EM) and pyroptosis (IL-1β, NLRP3, Caspase-1, GSDMD and GSDMD-N, by WB) in ulcers were evaluated.
RESULTS
After the treatment wound area rate, IL-1β by ELISA, and IL-1β, Caspase-1, GSDMD and GSDMD-N by WB of MSC-CM group were less than those of DM group. The thickness of the stratum granulosums of the skin, proliferation index of Ki67, mean optic density of CD31 and LC3B by IFC, and LC3B by WB of MSC-CM group were more than those of DM group. The present analysis demonstrated that the injection of MSC-CM into rats with DFUs enhanced the wound-healing process by accelerating wound closure, promoting cell proliferation and angiogenesis, enhancing cell autophagy, and reducing cell pyroptosis in ulcers.
CONCLUSIONS
Studies conducted indicate that MSC-CM administration could be a novel cell-free therapeutic approach to treat DFUs accelerating the wound healing process and avoiding the risk of living cells therapy.
目的
本研究旨在探讨骨髓间充质干细胞条件培养基（MSC-CM）治疗大鼠糖尿病足溃疡的作用。
方法
SD 大鼠给予高脂饮食联合腹腔注射 STZ 诱导 2 型糖尿病模型，糖尿病大鼠后腿手术制作糖尿病足溃疡模型。将大鼠分为四组（每组 n=6），即正常对照组（NC）、糖尿病对照组（DM-C）、MSC-CM 组和骨髓间充质干细胞组（MSCs）。MSC-CM 组于溃疡周围注射预处理大鼠骨髓 MSC 条件培养基，MSCs 组于溃疡周围注射大鼠骨髓 MSC，另两组注射 PBS。治疗后，观察创面愈合、再上皮化（皮肤颗粒层厚度，H&E 染色）、细胞增殖（Ki67，免疫组化）、血管生成（CD31，免疫荧光）、自噬（LC3B，免疫荧光和 WB；自噬溶酶体，电镜）和细胞焦亡（IL-1β、NLRP3、Caspase-1、GSDMD 和 GSDMD-N，WB）。
结果
治疗后，MSC-CM 组创面面积率、ELISA 检测的 IL-1β、WB 检测的 IL-1β、Caspase-1、GSDMD 和 GSDMD-N 均低于 DM 组，皮肤颗粒层厚度、Ki67 增殖指数、CD31 平均光密度、免疫荧光 LC3B 及 WB 检测 LC3B 均高于 DM 组。本研究表明，DFU 大鼠注射 MSC-CM 可通过加速创面闭合、促进细胞增殖和血管生成、增强细胞自噬、减少细胞焦亡来促进创面愈合。
结论
研究表明，MSC-CM 给药可能是一种新型无细胞治疗方法，可加速糖尿病足溃疡的愈合过程，避免活细胞治疗的风险。
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