微小RNA表达诱导巨噬细胞凋亡以响应(MRHO/IR/75/ER):一项体外和体内研究
MicroRNAs Expression Induces Apoptosis of Macrophages in Response to (MRHO/IR/75/ER): An In-Vitro and In-Vivo Study.
作者信息
Gholamrezaei Mostafa, Rouhani Soheila, Mohebali Mehdi, Mohammadi-Yeganeh Samira, Haji Molla Hoseini Mostafa, Haghighi Ali, Lasjerdi Zohreh, Hamidi Faezeh, Kazem Sharifi-Yazdi Mohammad
机构信息
Department of Parasitology and Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
出版信息
Iran J Parasitol. 2020 Oct-Dec;15(4):475-487. doi: 10.18502/ijpa.v15i4.4851.
BACKGROUND
We aimed to investigate the effect of miR-15a mimic and inhibitor of miR-155 expression on apoptosis induction in macrophages infected with Iranian strain of in-vitro and in-vivo.
METHODS
RAW 264.7 cells were infected with promastigotes (MRHO/IR/75/ER), and then were treated with miRNAs. For in-vivo experiment, BALB/c mice were inoculated with promastigotes, and then they were treated with miRNAs. For evaluation of miRNA therapeutic effect, in-vitro and in-vivo studies were performed using quantitative Real-time PCR, Flow cytometry, lesion size measurement, and Limiting Dilution Assay (LDA). This study was performed in Shahid Beheshti University of Medical Sciences in 2019.
RESULTS
In-vitro results of flow cytometry showed that using miR-15a mimic, miR-155 inhibitor or both of them increased apoptosis of macrophages. In in-vivo, size of lesion increased during experiment in control groups (<0.05) while application of both miR-155 inhibitor and miR-15a mimic inhibited the increase in the size of lesions within 6 wk of experiment (=0.85). LDA results showed that microRNA therapy could significantly decrease parasite load in mimic or inhibitor receiving groups compared to the control group (<0.05).
CONCLUSION
miR-155 inhibitor and miR-15a mimic in infected macrophages can induce apoptosis and reduce parasite burden. Therefore, miRNA-based therapy can be proposed as new treatment for cutaneous leishmaniasis.
背景
我们旨在研究miR-15a模拟物和miR-155表达抑制剂对体外和体内感染伊朗株利什曼原虫的巨噬细胞凋亡诱导的影响。
方法
RAW 264.7细胞用前鞭毛体(MRHO/IR/75/ER)感染,然后用miRNA处理。对于体内实验,BALB/c小鼠接种前鞭毛体,然后用miRNA处理。为评估miRNA的治疗效果,使用定量实时PCR、流式细胞术、病变大小测量和极限稀释分析(LDA)进行体外和体内研究。本研究于2019年在沙希德贝赫什提医科大学进行。
结果
流式细胞术的体外结果显示,使用miR-15a模拟物、miR-155抑制剂或两者均可增加巨噬细胞的凋亡。在体内,对照组实验期间病变大小增加(<0.05),而应用miR-155抑制剂和miR-15a模拟物在实验6周内均抑制了病变大小的增加(=0.85)。LDA结果显示,与对照组相比,接受模拟物或抑制剂的微RNA治疗组可显著降低寄生虫载量(<0.05)。
结论
利什曼原虫感染的巨噬细胞中的miR-155抑制剂和miR-15a模拟物可诱导凋亡并减轻寄生虫负担。因此,基于微RNA的治疗可作为皮肤利什曼病的新治疗方法。