The role of sub-transform of macrophages in renal ischemia/reperfusion injury in rats

doi:10.13459/j.cnki.cjap.2016.04.014

Abstract

Abstract: Objective:To investigate the role of sub-transform macrophage in ischemia/reperfusion renal injury in rats, as well as under-lying mechanisms. Methods:Thirty male Sprague-Dawley rats were randomly divided into ischemia/reperfusion (IRI, n=24, renal artery was occluded for 45 min) group and sham-operation (Sham, n=6) group. The kidneys in IRI group were collected at 0, 6, 24 and 72 h after operation (6 rats for each time point). The injury of the kidney was detected with HE staining. Immunohistochemistry staining was performed to evaluate the expression of proliferating cell nuclear antigen (PCNA). Real-time PCR was used to detect the mRNA expression of macrophage migration inhibitory factor (MIF). Moreover, the expression and location of MIF, monocyte chemoattractant protein-1 (MCP-1) and macrophage marker CD68 were examined by immunofluorescence staining. Most importantly, the distribution of macrophage subtypes M1 and M2 was analyzed by flow cytometry. Results:The worst pathologic damage of the renal tissues, as well as infiltration of inflammatory cells, was observed at 24 h after operation in IRI rats, with obvious recovery afterwards. Immunohistochemistry staining showed that the expression of PCNA was significantly increased after the ischemia/reperfusion, peaking at 6 h and reducing at 72 h after operation. Compared with sham group, the levels of MIF at mRNA and protein levels were both significantly increased after the ischemia/reperfusion, while the expression of MCP-1 was peaked at 6 h and decreased afterwards. Moreover, the expression of CD68-positive macrophages were significantly increased in IRI rats, with peaking at 24 h and reducing at 72 h. Furthermore, after 6 h of reperfusion, the percentage of M1 macrophages reached the peak, and thereafter the relative expression of M1 and M2 was reduced and increased, respectively. Conclusion:In the early phase of ischemia/per-fu sion renal injury, M1 macrophage results in renal damage, and afterwards the M2 macrophage is increased and repairs the renal damage by improving the cell proliferation.

Key words: kidney, ischemia/reperfusion, macrophage, injury and recovery

LIN Cheng-cheng, LU Hong, WU Lian-feng, LIANG Yong, CHEN Bi-cheng, BAI Yong-heng. The role of sub-transform of macrophages in renal ischemia/reperfusion injury in rats[J]. CJAP, 2016, 32(4): 338-342.

References

[1] Kucuk A, Kabadere S, Tosun M, et al. Protective effects of doxycycline in ischemia/reperfusion injury on kidney[J]. J Physiol Biochem, 2009, 65(2):183-191.
[2] Liangos O, Tighiouart H, Perianayagam MC, et al. Compar-ative analysis of urinary biomarkers for early detection of acute kidney injury following cardiopulmonary bypass[J]. Biomarkers, 2009, 14(6):423-431.
[3] Humphreys BD, Valerius MT, Kobayashi A, et al. Intrinsic epithelial cells repair the kindey after injury[J]. Cell Stem Cell, 2008, 2(3):284-291.
[4] 王玉, 李晓玫. 肾脏固有细胞的表型转化特征及其病理生理意义[J]. 中国病理生理杂志, 2002, 18(10):1303-1307.
[5] Lee S, Huen S, Nishio H, et al. Distinct macrophage phe-notypes contribute to kindey injury and repair[J]. J Am Soc Nephrol, 2011, 22(2):317-326.
[6] 赵雅妮, 李惊子, 王海燕. 巨噬细胞在肾脏损伤中的作用[J]. 中国病理生理杂志, 2004, 20(11):2147-2150.
[7] Perico N, Cattaneo D, Sayegh MH, et al. Delayed graft fu nction in kidney transplantation[J]. Lancet, 2004, 364(9447):1814-1827.
[8] 魏日胞, 李平, 丁瑞, 等. 葡天胶囊对大鼠肾脏缺血再灌注损伤的保护作用[J]. 中国中西医结合肾病杂志, 2010, 11(4):295-297.
[9] Nelson PJ, Rees AJ, Griffin MD, et al. The renal mononu-clear phagocytic system[J]. J Am Soc Nephrol, 2012, 23(2):194-203.
[10] 赵春玲, 张春来, 李莉华, 等. IL-1受体拮抗剂减轻离体猪肾缺血/再灌注损伤的作用及其机制[J]. 中国应用生理学杂志, 2001, 17(3):247-250.
[11] 张翠萍, 谢印芝, 陈鹏, 等. 低氧对巨噬细胞分泌TNF-α和IL-6的影响及其机制[J]. 中国应用生理学杂志, 2005, 21(3):281-284.
[12] Foote A, Briganti EM, Kipen Y, et al. Macrophage migra-tion inhibitory factor in systemic lupus erythematosus[J]. J Rheumatol, 2004, 31(2):268-273.
[13] 杨立, 秦大山, 岳中瑾. 肾缺血再灌注中单核细胞趋化蛋白-1的表达与细胞凋亡的相关作用[J]. 第四军医大学学报, 2003, 24(18):1659-1661.
[14] Cao Q, Wang Y, Zheng D, et al. IL-10/TGF-beta-modified macrophages induce regulatory T cells and protect against adriamycin nephrosis[J]. J Am Soc Nephrol, 2010, 21(6):933-942.
[15] 魏盼, 黄文娟, 万辛, 等. 巨噬细胞在小鼠肾脏缺血/再灌注损伤修复中的作用[J]. 中华肾脏病杂志, 2014, 30(10):757-762.