Rahbarghazi Reza, Keyhanmanesh Rana, Rezaie Jafar, Mirershadi Fatemeh, Heiran Hossain, Saghaei Bagheri Hesam, Saberianpour Shirin, Rezabakhsh Aysa, Delkhosh Aref, Bagheri Yasin, Rajabi Hadi, Ahmadi Mahdi
Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Department of Applied Cell Sciences, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran.
Iran J Basic Med Sci. 2021 Mar;24(3):369-376. doi: 10.22038/ijbms.2021.49008.11231.
There are still challenges regarding c-kit+ cells' therapeutic outcome in the clinical setting. Here, we examined the c-kit+ cell effect on the alleviation of asthma by modulating miRNAs expression.
To induce asthma, male rats were exposed to ovalbumin. Bone marrow-derived c-kit+ cells were enriched by MACS. Animals were classified into four groups (6 rats each). Control rats received PBS intratracheally; Ovalbumin-sensitized rats received PBS intratracheally; Ovalbumin-sensitized rats received PBS intratracheally containing 3×105 c-kit+ and c-kit- cells. Cells were stained with Dil fluorescent dye to track in vivo condition. Pathological changes were monitored in asthmatic rats after transplantation of c-kit+ and c-kit- cells. Serum levels of IL-4 and INF-γ were measured by ELISA. Transcription of miRNAs (-126 and 133) was assessed by real-time PCR analysis.
Pathological examination and Th1 and Th2 associated cytokine fluctuation confirmed the occurrence of asthma in rats indicated by chronic changes and prominent inflammation compared with the control group (<0.05). Both c-kit+ and c-kit- cells were verified in pulmonary niche. Administration of c-kit positive cells had the potential to change INF-γ/IL-4 ratio close to the normal values compared with matched-control asthmatic rats (<0.05). We also found that c-kit+ cells regulated the expression of miRNA-126 and -133, indicated by an increase of miRNA-133 and decrease of miRNA-126 compared with cell-free sensitized groups (<0.05).
c-kit- cells were unable to promote any therapeutic outcomes in the asthmatic milieu. c-kit+ cells had the potential to diminish asthma-related pathologies presumably by controlling the transcription of miRNA-126 and -133.
在临床环境中，c-kit+细胞的治疗效果仍存在挑战。在此，我们通过调节微小RNA（miRNA）的表达来研究c-kit+细胞对哮喘缓解的影响。
为诱导哮喘，将雄性大鼠暴露于卵清蛋白。通过磁珠分选法富集骨髓来源的c-kit+细胞。动物被分为四组（每组6只大鼠）。对照组大鼠经气管内给予磷酸盐缓冲液（PBS）；卵清蛋白致敏大鼠经气管内给予PBS；卵清蛋白致敏大鼠经气管内给予含3×105个c-kit+细胞和c-kit-细胞的PBS。细胞用Dil荧光染料染色以追踪其体内情况。在移植c-kit+细胞和c-kit-细胞后，监测哮喘大鼠的病理变化。通过酶联免疫吸附测定（ELISA）测量血清白细胞介素-4（IL-4）和干扰素-γ（INF-γ）水平。通过实时聚合酶链反应（PCR）分析评估miRNA（-126和133）的转录情况。
病理检查以及Th1和Th2相关细胞因子的波动证实，与对照组相比，大鼠出现了由慢性变化和明显炎症所表明的哮喘（<0.05）。在肺微环境中证实了c-kit+细胞和c-kit-细胞的存在。与匹配的对照哮喘大鼠相比，给予c-kit阳性细胞有可能使INF-γ/IL-4比值接近正常值（<0.05）。我们还发现，与无细胞致敏组相比，c-kit+细胞调节了miRNA-126和-133的表达，表现为miRNA-133增加而miRNA-126减少（<0.05）。
在哮喘环境中，c-kit-细胞无法促进任何治疗效果。c-kit+细胞有可能通过控制miRNA-126和-133的转录来减轻与哮喘相关的病理变化。
