Comparative Study of Effect of Poly I:C on PKR/eIF2α Signal Path in Naked Mole-rats and Mouse Macrophages

doi:10.3969/j.issn.1674-5817.2019.06.009

Abstract

Abstract: Objective To compare the affect of polyinosinic-polycytidylic acid(Poly I:C) on double-stranded RNA-dependent protein kinase R (PKR)/eukaryotic initiation factor-2α (EIF2α) signaling pathway in macrophages of naked mole-rats and mouse. Methods The naked mole-rat and C57BL/6J mice which physiological age-matched were randomly divided into control group, Poly I:C administration group, 2,6-Diaminopurine (2-AP) administration group and Poly I:C+2-AP administration group. After administration, the expression of phosphorylated PKR (Pho-PKR), PKR, phosphorylated EIF2α (Pho-EIF2α), EIF2α and Caspase-8 protein in the cells were detected by Western blotting. Results After Poly I:C administration, the phosphorylation levels of PKR and eIF2α in mice macrophages were significantly decreased (P<0.01), while those in naked mole-rat were significantly increased (P<0.01).This phenomelon indicated that Poly I:C could inhibit PKR activity in mice, but activite PKR in naked mole-rat; Poly I:C and 2-AP intervention group showed that PKR and phosphorylated PKR were down-regulated both in mice and naked mole-rats (P<0.01), especially PKR in naked mole-rats. Active PKR and active eIF2α was significantly decreased (P<0.01) in mouse macrophages, after 2-AP administration. Active PKR in naked mole-rat was significantly decreased (P<0.01) too. After inhibition of PKR activity by 2-AP, Caspase-8 expression levels in naked mole-rat were significantly reduced (P<0.01), but there were no significant differences in mouse cells. Conclusion Compared to mouse, naked mole-rat's cells were highly resistant to Poly I:C stimulation,PKR in naked mole-rats was more sensitive to 2-AP, PKR activity in naked-mole rat cells promoted the expression of Caspase-8.

Key words: Naked mole-rat, Polyinosinic-polycytidylic acid (Poly I:C), Double-stranded RNA-dependent protein kinase R (PKR), Caspase-8

CLC Number:

Q95-33

LIN Li-fang, ZHANG Cheng-cai, LI Yu, ZHANG Qian-qian, CHEN Chao, LI Lei-cheng, YANG Wen-jing, LI Li, CUI Shu-fang. Comparative Study of Effect of Poly I:C on PKR/eIF2α Signal Path in Naked Mole-rats and Mouse Macrophages[J]. Laboratory Animal and Comparative Medicine, 2019, 39(6): 473-478.

References

[1] Edrey YH, Park TJ, Kang H, et al.Endocrine function and neurobiology of the longest-living rodent, the naked mole-rat[J]. Exp Gerontol, 2011, 46(2-3):116-123.
[2] Buffenstein R.Negligible senescence in the longest living rodent, the naked mole-rat: insights from a successfully aging species[J]. J Comp Physiol B, 2008, 178(4):439-445.
[3] Lewis KN, Mele J, Hornsby PJ, et al.Stress resistance in the naked mole-rat: the bare essentials a mini-review[J]. Gerontology, 2012, 58(5):453-462.
[4] Salmon AB, Sadighi Akha AA, Buffenstein R, et al.Fibroblasts from naked mole-rats are resistant to multiple forms of cell injury, but sensitive to peroxide, ultraviolet light, and endoplasmic reticulum stress[J]. J Gerontol A Biol Sci Med Sci, 2008, 63(3):232-241.
[5] 林丽芳, 肖邦, 赵善民, 等. Poly I:C刺激对裸鼹鼠体内自噬水平的影响[J]. 实验动物与比较医学, 2014, 34(5):411-416.
[6] Arredouani MS, Palecanda A, Koziel H, et al.MARCO is the major binding receptor for unopsonized particles and bacteria on human alveolar macrophages[J]. Immunol, 2005,75(9):6058-6064.
[7] Swanson JA, Baer SC.Phagocytosis by zippers and triggers[J]. Trends Cell Biol, 1995, 3:89-93.
[8] 蒯守刚, 俞用芳, 朱启贯. 巨噬细胞研究进展[J]. 中国实验诊断, 2008, 12(2):266-272.
[9] Soilleux EJ, Morris LS, Leslie G, et al.Constitutive and induced expression of DC-SIGN on dendritic cell and macrophage subpopulations in situ and in vitro[J]. J Leukoc Biol, 2002, 71(3):445-457.
[10] Relloso M, Puig-Kröger A, Pello OM, et al.DC-SIGN (CD209) expression is IL-4 dependent and is negatively regulated by IFN,TGF-beta, and anti-inflammatory agents[J]. J Immunol, 2002, 168(6):2634-2643.
[11] Arrode-Brusés G, Brusé JL.Maternal immune activation by poly I:C induces expression of cytokines IL-1beta and IL-13, chemokine MCP-1 and colony stimulating factor VEGF in fetal mouse brain[J]. J Neuroinflammation, 2012, 9:83.
[12] Fortier ME, Kent S, Ashdown H, et al.The viral mimic, polyinosinic:polycytidylic acid, induces fever in rats via an interleukin-1-dependent mechanism[J]. Am J Physiol Regul Integr Comp Physiol, 2004, 287(4):759-766.
[13] Nanduri S, Rahman F, Williams BR, et al.A dynamically tuned double-stranded RNA binding mechanism for the activation of antiviral kinase PKR[J]. EMBO J, 2000, 19(20):5567-5574.
[14] Levin DH, Petryshyn R, London IM.Characterization of double-stranded-RNA-activated kinase that phosphorylates alpha subunit of eukaryotic initiation factor 2(Eif-2 alpha) in reticulacyte lysates[J]. Proc Natl Acad Sci U S A, 1980, 77(2):832-836.
[15] Elia A, Vyas J, Laing KG, et al.Ribosomal protein L22 inhibits regulation of cellular activities by the Epstein-Barr virus small RNA EBER-1[J]. Eur J Biochem, 2004, 271(10):1895-1905.