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[补体C5a对三氯乙烯致敏小鼠免疫性肾损伤中MCP-1和NGAL表达的影响]

[Effect of complement C5a on the expression of MCP-1 and NGAL in immune kidney injury of trichloroethylene sensitized mice].

作者信息

Huang L P, Wang F, Dai Y Y, Xu Q Y, Zhang J X, Zhu Q X

机构信息

Department of Occupational and Environmental Health, School of Public Health, Anhui Medical University, Hefei 230032, China.

Institute of Dermatology, the First Affiliated Hospital of Anhui Medical University, Hefei 230032, China.

出版信息

Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2020 Mar 20;38(3):161-167. doi: 10.3760/cma.j.cn121094-20190717-00310.

DOI:10.3760/cma.j.cn121094-20190717-00310
PMID:32306687
Abstract

To explore the possible role of C5a in the pathogenesis of renal injury in TCE- sensitized mice, to analyze the impact of expression of neutrophil gelatinase-associated lipocalin (NGAL) and monocyte chemotactic protein-1 (MCP-1) in the presence or absence of C5a receptor antagonist (C5aRA) pretreatment. A total of 50 female specific pathogens free(SPF) BALB/c mice were randomly divided into blank control group (=5) , solvent control group (=5) , TCE group (=20) , and TCE+C5aRA group (= 20) . After one week for adaptive feeding, a mouse model of TCE-induced skin sensitization was established by treating with 50% TCE and 30% TCE in turn. The mice in solvent control group accept same reagents without TCE and the mice in blank control group underwent nothing. In TCE +C5aRA group, except for the TCE solution treatment, mice were intraperitoneally injected with 0.5 mg/kg C5aRA solution at the time of challenge. And the skin erythema and edema reaction were scored 24 h after the last challenge. The mice were divided into sensitization positive group and sensitization negative group according to the scoring result. The mice were aseptically sacrificed 72 h after the last challenge to obtain the kidneys. The structural damage of kidney was observed after histopathological staining. The levels of NGAL and MCP-1 mRNA and proteins were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry (IHC) , respectively. The sensitization rate of mice in TCE group and TCE+C5aRA group was 45.0% (9/20) and 40.0% (8/20) , respectively. No skin lesions was found in the mice of blank control group and solvent control group. The results of histopathological staining showed that the TCE sensitization positive mice showed renal tubular dilatation, vacuolar degeneration of renal tubular epithelial cells, and infiltration of interstitial cells. The pathological damage of the kidney in TCE sensitization positive group was mild, and no inflammatory cell infiltration was seen. The data of qRT-PCR showed that the expression levels of NGAL and MCP-1 mRNA in the TCE sensitization positive group were significantly increased than in solvent control group and TCE sensitization negative group (<0.05) , while the levels of NGAL and MCP-1 mRNA in TCE+C5aRA sensitization positive group were decreased than TCE sensitization positive group ( <0.05) . The results of IHC showed that the expression levels of NGAL and MCP-1 in TCE protein sensitization positive group were significantly higher than those in solvent control group and TCE sensitization negative group (<0.05) . After C5aRA pretreatment, the expression levels of NGAL and MCP-1 protein were decreased than the mice in TCE sensitization positive group (<0.05) . The regulation of C5a on the expression of MCP-1 and NGAL may participate in TCE- induced mice kidney damage, and pharmacological inhibition of C5a seems to be an effective way to protect the kidney injury in TCE-sensitized mice.

摘要

为探讨C5a在三氯乙烯(TCE)致敏小鼠肾损伤发病机制中的可能作用,分析在存在或不存在C5a受体拮抗剂(C5aRA)预处理的情况下中性粒细胞明胶酶相关脂质运载蛋白(NGAL)和单核细胞趋化蛋白-1(MCP-1)表达的影响。将50只雌性无特定病原体(SPF)BALB/c小鼠随机分为空白对照组(n = 5)、溶剂对照组(n = 5)、TCE组(n = 20)和TCE + C5aRA组(n = 20)。适应性饲养1周后,依次用50% TCE和30% TCE处理建立TCE诱导的皮肤致敏小鼠模型。溶剂对照组小鼠接受不含TCE的相同试剂,空白对照组小鼠不做任何处理。在TCE + C5aRA组中,除TCE溶液处理外,小鼠在激发时腹腔注射0.5 mg/kg C5aRA溶液。在最后一次激发后24小时对皮肤红斑和水肿反应进行评分。根据评分结果将小鼠分为致敏阳性组和致敏阴性组。在最后一次激发后72小时无菌处死小鼠获取肾脏。组织病理学染色后观察肾脏的结构损伤。分别通过定量实时聚合酶链反应(qRT-PCR)和免疫组织化学(IHC)检测NGAL和MCP-1 mRNA及蛋白水平。TCE组和TCE + C5aRA组小鼠的致敏率分别为45.0%(9/20)和40.0%(8/20)。空白对照组和溶剂对照组小鼠未发现皮肤病变。组织病理学染色结果显示,TCE致敏阳性小鼠表现为肾小管扩张、肾小管上皮细胞空泡变性和间质细胞浸润。TCE致敏阳性组肾脏的病理损伤较轻,未见炎性细胞浸润。qRT-PCR数据显示,TCE致敏阳性组中NGAL和MCP-1 mRNA的表达水平显著高于溶剂对照组和TCE致敏阴性组(P < 0.05),而TCE + C5aRA致敏阳性组中NGAL和MCP-1 mRNA的水平低于TCE致敏阳性组(P < 0.05)。IHC结果显示,TCE蛋白致敏阳性组中NGAL和MCP-1的表达水平显著高于溶剂对照组和TCE致敏阴性组(P < 0.05)。C5aRA预处理后,NGAL和MCP-1蛋白的表达水平低于TCE致敏阳性组小鼠(P < 0.05)。C5a对MCP-1和NGAL表达的调节可能参与TCE诱导的小鼠肾损伤,对C5a的药理学抑制似乎是保护TCE致敏小鼠肾损伤的有效方法。

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