小鼠胸腺碎片中的细胞呼吸。

Cellular Respiration in Thymic Fragments from Mice.

机构信息

Department of Microbiology and Immunology, College of Medicine and Health Sciences, UAE University, 15551 Al Ain, Abu Dhabi, United Arab Emirates.

Department of Pediatrics, College of Medicine and Health Sciences, UAE University, 15551 Al Ain, Abu Dhabi, United Arab Emirates.

出版信息

Front Biosci (Landmark Ed). 2022 Jul 29;27(8):230. doi: 10.31083/j.fbl2708230.

DOI:10.31083/j.fbl2708230

PMID:36042174

Abstract

BACKGROUND

This report aims to detail the use of the phosphorescence oxygen analyzer for investigation of thymic responses to pharmaceutical agents, in particular immunosuppressants and immunomodulators. Sirolimus (a highly specific inhibitor of the 'molecular target of rapamycin', mTOR) and ozanimod (an agonist of the sphingosine 1-phosphate receptor, recently approved for treatment of multiple sclerosis and ulcerative colitis) are used for this purpose.

METHODS

Thymic fragments from mice were placed in glass vials containing phosphate-buffered saline, bovine albumin, and Pd(II) meso-tetra (sulfophenyl) tetrabenzoporphyrin. The vials were sealed from air, and the cellular oxygen consumption was monitored as function of time.

RESULTS

The decline of dissolved oxygen concentration with time (d[O2]/d) was linear; thus, its rate (thymocyte respiration) was expressed as μM O2 min-1. Cyanide inhibited respiration, confirming the oxygen consumption was in cytochrome oxidase. In age-matched mice, the rate of thymocyte respiration (mean ± SD, in μM O2 min-1 mg-1) was 0.046 ± 0.011 (median = 0.043, range = 0.028 to 0.062, n = 10). In thymic fragments from littermates, this rate was inhibited in the presence of sirolimus (16% lower) or ozanimod (29% lower).

CONCLUSIONS

Thymocyte respiration can serve as a surrogate biomarker for studying the mode-of-action and the cytotoxicity of immunotoxins and immunosuppressants.

摘要

背景

本报告旨在详细介绍磷光氧分析仪在研究药物对胸腺的作用，特别是免疫抑制剂和免疫调节剂方面的应用。本研究使用了西罗莫司（一种高度特异性的雷帕霉素“分子靶标”抑制剂，mTOR）和奥扎尼莫德（一种鞘氨醇 1-磷酸受体激动剂，最近被批准用于治疗多发性硬化症和溃疡性结肠炎）。

方法

将来自小鼠的胸腺片段置于含有磷酸盐缓冲盐水、牛白蛋白和 Pd(II)meso-四（磺苯基）四苯并卟啉的玻璃小瓶中。小瓶与空气隔绝，监测细胞耗氧量随时间的变化。

结果

溶解氧浓度随时间的下降（d[O2]/d）呈线性关系；因此，其速率（胸腺细胞呼吸）表示为μM O2 min-1。氰化物抑制呼吸，证实氧消耗发生在细胞色素氧化酶中。在年龄匹配的小鼠中，胸腺细胞呼吸的速率（平均值±SD，以μM O2 min-1 mg-1表示）为 0.046±0.011（中位数=0.043，范围=0.028 至 0.062，n=10）。在同窝仔鼠的胸腺片段中，西罗莫司（降低 16%）或奥扎尼莫德（降低 29%）存在时，该速率受到抑制。