-
摘要:
目的 探讨白细胞介素（IL）-17C在小鼠肾移植中的作用及其机制。 方法 以Balb/c（H-2Kd）小鼠为供体，IL-17C基因敲除（IL-17CKO）小鼠（敲除组）、C57BL/6J（H-2Kb）小鼠（野生组）为受体，建立小鼠生命支持型肾移植模型。术后比较两组小鼠的体质量及存活时间。采用苏木素-伊红（HE）染色及过碘酸-雪夫（PAS）染色对移植肾进行病理学检查。采用逆转录聚合酶链反应（RT-PCR）检测移植肾组织中颗粒酶B、干扰素（IFN）-γ、肿瘤坏死因子（TNF）-α、IL-6及IL-1β的信使核糖核酸（mRNA）表达水平，采用流式细胞术检测移植肾组织中炎症细胞浸润情况。 结果 移植术后敲除组小鼠存活时间显著短于野生组小鼠（P=0.031），且体质量下降程度更明显，但差异无统计学意义。病理分析发现敲除组小鼠移植肾损伤较野生组小鼠明显加重。敲除组小鼠移植肾组织中颗粒酶B、IFN-γ、TNF-α、IL-6的mRNA表达水平均显著高于野生组（均为P < 0.01），IL-1β mRNA表达呈降低趋势，但差异无统计学意义（P=0.16）。流式细胞分析发现敲除组小鼠移植肾组织中CD45+CD11b+Ly6G+中性粒细胞和CD45+CD11b+Ly6Chi单核细胞浸润较野生组小鼠明显增加（分别为P < 0.05，P < 0.01），而CD45+ Ly6Chi F4/80+巨噬细胞浸润无明显变化（P > 0.05）。 结论 IL-17C参与肾移植后炎症反应的调控，可能通过降低促炎细胞因子的表达及炎症细胞的浸润来减轻急性排斥反应，改善移植肾存活情况。 Abstract:Objective To investigate the effect and mechanism of interleukin (IL)-17C in mice undergoing kidney transplantation. Methods The life-supporting kidney transplantation mice models were established using Balb/c (H-2Kd) mice as the donors, IL-17C gene knock out (IL-17CKO) mice (knockout group) and C57BL/6J(H-2Kb) mice (wild group) were chosen as the recipients. The postoperative body mass and survival time of mice were statistically compared between two groups. Pathological examination of the kidney graft was performed by using hematoxylin-eosin (HE) staining and periodic acid-Schiff (PAS) staining. The expression levels of granzyme B, interferon (IFN)-γ, tumor necrosis factor (TNF)-α, IL-6 and IL-1β messenger ribonucleic acid (mRNA) in the kidney graft tissue were quantitatively measured by reverse transcription polymerase chain reaction (RT-PCR). The proportion of inflammatory cell infiltration in the kidney graft tissue was detected by flow cytometry. Results In the knockout group, the survival time of mice after kidney transplantation was significantly shorter than that of the wild mice (P=0.031). The body mass was more evidently decreased in the knockout group with no statistical significance from that in the wild group. Pathological examination demonstrated that the kidney graft injury in the knockout group was significantly worse than that in the wild group. The mRNA expression levels of granzyme B, IFN-γ, TNF-α, IL-6 mRNA in the knockout group were significantly up-regulated compared with those in the wild group (all P < 0.01). The mRNA expression level of IL-1β showed a decreasing trend with no statistical significance (P=0.16). Flow cytometry analysis revealed that the infiltration of CD45+CD11b+Ly6G+ neutrophil and CD45+CD11b+Ly6Chi monocyte in the kidney graft of knockout mice was significantly higher compared with that of the wild mice (P < 0.05, P < 0.01), whereas the infiltration of CD45+Ly6ChiF4/80+ macrophage did not significantly differ between two groups (P > 0.05). Conclusions IL-17C participates in the regulation of inflammatory response after kidney transplantation. It can alleviate acute rejection and improve the survival of kidney graft by down-regulating the expression of pro-inflammatory cytokines and infiltration of inflammatory cells. -
图 3 小鼠移植肾病理学改变（HE，×200）
注：与野生组小鼠比较，敲除组小鼠肾间质可见大量炎症细胞浸润、血管纤维化及周围炎症浸润，同时伴有血栓形成。
Figure 3. The pathological changes of the kidney allograft in mice
图 4 小鼠移植肾病理学改变（PAS，×200）
注：与野生组小鼠比较，敲除组小鼠移植肾的肾小管细胞刷状缘明显减少甚至消失。
Figure 4. The pathological changes of the kidney allograft in mice
图 5 小鼠移植肾组织匀浆中炎症细胞因子表达情况
注：与野生组小鼠比较，aP < 0.01。
Figure 5. Expression of inflammatory cytokines in renal allograft homogenate of mice
-
[1] TAJIMA S, FU R, SHIGEMATSU T, et al. Urinary human epididymis secretory protein 4 as a useful biomarker for subclinical acute rejection three months after kidney transplantation[J]. Int J Mol Sci, 2019, 20(19):E4699. DOI: 10.3390/ijms20194699. [2] SALCEDO-HERRERA S, PINTO RAMIREZ JL, GARCíA-LOPEZ A, et al. Acute rejection in kidney transplantation and early beginning of tacrolimus[J]. Transplant Proc, 2019, 51(6):1758-1762. DOI: 10.1016/j.transproceed.2019.04.048. [3] HU X, SU M, LIN J, et al. Corin is downregulated in renal ischemia/reperfusion injury and is associated with delayed graft function after kidney transplantation[J]. Dis Markers, 2019:9429323. DOI: 10.1155/2019/9429323. [4] SU S, ZHANG P, ZHANG Q, et al. GSK-3β inhibitor induces expression of the TLR4/MyD88/NF-κB signaling pathway to protect against renal ischemia-reperfusion injury during rat kidney transplantation[J]. Inflammation, 2019, 42(6):2105-2118.DOI: 10.1007/s10753-019-01074-2. [5] 金钟大, 具春花.大鼠心肾联合移植模型的建立[J].实用医学杂志, 2017, 33(16):2639-2642. DOI: 10.3969/j.issn.1006-5725.2017.16.008.JIN ZD, JU CH. Establishment of a rat model of heart and kidney transplantation[J]. J Pract Med, 2017, 33(16):2639-2642.DOI: 10.3969/j.issn.1006-5725.2017.16.008. [6] CHANG SH, REYNOLDS JM, PAPPU BP, et al. Interleukin-17C promotes Th17 cell responses and autoimmune disease via interleukin-17 receptor E[J]. Immunity, 2011, 35(4):611-621. DOI: 10.1016/j.immuni. 2011.09.010. [7] SONG X, ZHU S, SHI P, et al. IL-17RE is the functional receptor for IL-17C and mediates mucosal immunity to infection with intestinal pathogens[J]. Nat Immunol, 2011, 12(12):1151-1158. DOI: 10.1038/ni.2155. [8] LI H, CHEN J, HUANG A, et al. Cloning and characterization of IL-17B and IL-17C, two new members of the IL-17 cytokine family[J]. Proc Natl Acad Sci U S A, 2000, 97(2):773-778. doi: 10.1073/pnas.97.2.773 [9] KROHN S, NIES JF, KAPFFER S, et al. IL-17C/IL-17 receptor E signaling in CD4+ T cells promotes Th17 cell-driven glomerular inflammation[J]. J Am Soc Nephrol, 2018, 29(4):1210-1222. DOI: 10.1681/ASN.2017090949. [10] AGGARWAL S, GHILARDI N, XIE MH, et al. Interleukin-23 promotes a distinct CD4 T cell activation state characterized by the production of interleukin-17 [J]. J Biol Chem, 2003, 278(3):1910-1914. doi: 10.1074/jbc.M207577200 [11] LOVERRE A, DIVELLA C, CASTELLANO G, et al. T helper 1, 2 and 17 cell subsets in renal transplant patients with delayed graft function[J]. Transpl Int, 2011, 24(3):233-242. DOI: 10.1111/j.1432-2277.2010.01157.x. [12] TURNER JE, PAUST HJ, STEINMETZ OM, et al. The Th17 immune response in renal inflammation[J]. Kidney Int, 2010, 77(12):1070-1075. DOI: 10.1038/ki.2010.102. [13] HAOUAMI Y, DHAOUADI T, SFAR I, et al. The role of IL-23/IL-17 axis in human kidney allograft rejection[J]. J Leukoc Biol, 2018, 104(6):1229-1239. DOI: 10.1002/JLB.5AB0318-148R. [14] LOVERRE A, TATARANNI T, CASTELLANO G, et al. IL-17 expression by tubular epithelial cells in renal transplant recipients with acute antibody-mediated rejection[J]. Am J Transplant, 2011, 11(6):1248-1259. DOI: 10.1111/j.1600-6143.2011.03529.x. [15] LI J, BASLER M, ALVAREZ G, et al. Immunoproteasome inhibition prevents chronic antibody-mediated allograft rejection in renal transplantation[J]. Kidney Int, 2018, 93(3):670-680. DOI: 10.1016/j.kint. 2017.09.023. [16] HWANG SY, KIM HY. Expression of IL-17 homologs and their receptors in the synovial cells of rheumatoid arthritis patients[J]. Mol Cells, 2005, 19(2):180-184. http://cn.bing.com/academic/profile?id=31356686141710a81df0f65a4130c149&encoded=0&v=paper_preview&mkt=zh-cn [17] BUTCHER MJ, WASEEM TC, GALKINA EV. Smooth muscle cell-derived interleukin-17C plays an atherogenic role via the recruitment of proinflammatory interleukin-17A+ T cells to the aorta[J]. Arterioscler Thromb Vasc Biol, 2016, 36(8):1496-1506. DOI: 10.1161/ATVBAHA. 116.307892. [18] RAMIREZ-CARROZZI V, SAMBANDAM A, LUIS E, et al. IL-17C regulates the innate immune function of epithelial cells in an autocrine manner[J]. Nat Immunol, 2011, 12(12):1159-1166. DOI: 10.1038/ni.2156. [19] TANAKA S, NAGASHIMA H, CRUZ M, et al. Interleukin-17C in human helicobacter pylori gastritis[J]. Infect Immun, 2017, 85(10):e00389-17. DOI: 10.1128/IAI.00389-17. [20] REYNOLDS JM, MARTINEZ GJ, NALLAPARAJU KC, et al. Cutting edge: regulation of intestinal inflammation and barrier function by IL-17C[J]. J Immunol, 2012, 189(9):4226-4230. DOI: 10.4049/jimmunol.1103014. [21] JAMIESON KC, TRAVES SL, KOOI C, et al. Rhinovirus and bacteria synergistically induce IL-17C release from human airway epithelial cells to promote neutrophil recruitment[J].J Immunol, 2018, 202(1): 160-170. DOI: 10.4049/jimmunol.1800547. [22] HIROTA K, YOSHITOMI H, HASHIMOTO M, et al. Preferential recruitment of CCR6-expressing Th17 cells to inflamed joints via CCL20 in rheumatoid arthritis and its animal model[J]. J Exp Med, 2007, 204(12):2803-2812. doi: 10.1084/jem.20071397 [23] WANG JC, LEE JY, CHRISTIAN S, et al. The Rap1-cofilin-1 pathway coordinates actin reorganization and MTOC polarization at the B cell immune synapse[J]. J Cell Sci, 2017, 130(6):1094-1109. DOI: 10.1242/jcs.191858. [24] IBAÑEZ-VEGA J, DEL VALLE BATALLA F, SAEZ JJ, et al. Proteasome dependent actin remodeling facilitates antigen extraction at the immune synapse of B cells[J]. Front Immunol, 2019, 10:225. DOI: 10.3389/fimmu.2019.00225. [25] GONG H, MA S, LIU S, et al. IL-17C mitigates murine acute graft-vs.-host disease by promoting intestinal barrier functions and treg differentiation[J]. Front Immunol, 2018, 9:2724. DOI: 10.3389/fimmu.2018.02724. -
下载:
点击查看大图
图(6)
计量
- 文章访问数: 190
- HTML全文浏览量: 106
- PDF下载量: 16
- 被引次数: 0
