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PTX3对重症急性胰腺炎小鼠肾损伤的作用及机制研究
Effect and mechanism of PTX3 on kidney injury in mice with severe acute pancreatitis
- 局解手术学杂志 2023年32卷第7期 页码：561-566
- 作者机构：
海南西部中心医院重症医学科，海南 儋州 571799
- 作者简介：
赵展庆，E-mail：1834163815@qq.com
- 基金信息：海南省自然科学基金面上项目(820MS154)
DOI：10.11659/jjssx.07E022071
中图分类号： R657.5+1纸质出版日期：2023-07-25，
收稿日期：2022-07-19，
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李川,钟育武,赵展庆.PTX3对重症急性胰腺炎小鼠肾损伤的作用及机制研究[J].局解手术学杂志,2023,32(07):561-566.
LI Chuan,ZHONG Yu-wu,ZHAO Zhan-qing.Effect and mechanism of PTX3 on kidney injury in mice with severe acute pancreatitis[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2023,32(07):561-566.
李川,钟育武,赵展庆.PTX3对重症急性胰腺炎小鼠肾损伤的作用及机制研究[J].局解手术学杂志,2023,32(07):561-566. DOI： 10.11659/jjssx.07E022071.
LI Chuan,ZHONG Yu-wu,ZHAO Zhan-qing.Effect and mechanism of PTX3 on kidney injury in mice with severe acute pancreatitis[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2023,32(07):561-566. DOI： 10.11659/jjssx.07E022071.
摘要
目的
2
探究PTX3对重症急性胰腺炎（SAP）小鼠肾损伤的作用及其可能的机制。
方法
2
40只小鼠随机分为空白对照组、SAP组、PTX3组和anti-PTX3组，每组10只。除空白对照组外，其余各组小鼠腹腔注射20%左旋精氨酸建立SAP小鼠模型。PTX3组和anti-PTX3组小鼠分别腹腔注射重组小鼠PTX3（0.5 mg/kg）和PTX3单克隆抗体（0.2 mg/kg），空白对照组和SAP组小鼠注射等量生理盐水，24 h后处死所有小鼠。全自动生化分析仪检测血清淀粉酶（AMY）、肌酐（Cr）和尿素氮（BUN）水平；HE染色观察小鼠胰腺和肾组织病理学变化；TUNEL染色检测肾组织细胞凋亡情况；ELISA检测血清白细胞介素（IL-6）、肿瘤坏死因子-α（TNF-α）、IL-4和IL-13水平；Western blot检测PTX3、iNOS、CD16/32、CD206和Arg-1蛋白表达；流式细胞术检测肾组织中M1型和M2型巨噬细胞比例。
结果
2
与空白对照组相比，SAP组小鼠AMY、BUN、Cr、IL-6、TNF-α水平及胰腺组织病理学评分、肾小管损伤评分、细胞凋亡率、M1型巨噬细胞比例、PTX3蛋白表达、iNOS和CD16/32蛋白表达明显升高（
P
<
0.05），IL-4和IL-13水平、CD206和Arg-1蛋白表达、M2型巨噬细胞比例明显降低（
P
<
0.05）；与SAP组相比，PTX3组小鼠AMY、BUN、Cr、IL-6、TNF-α水平及胰腺组织病理学评分、肾小管损伤评分、细胞凋亡率、M1型巨噬细胞比例、PTX3蛋白表达、iNOS和CD16/32蛋白表达明显升高（
P
<
0.05），IL-4和IL-13水平、CD206和Arg-1蛋白表达、M2型巨噬细胞比例明显降低（
P
<
0.05）；与SAP组相比，anti-PTX3组小鼠AMY、BUN、Cr、IL-6、TNF-α水平及胰腺组织病理学评分、肾小管损伤评分、细胞凋亡率、M1型巨噬细胞比例、PTX3蛋白表达、iNOS和CD16/32蛋白表达明显降低（
P
<
0.05），IL-4和IL-13水平、CD206和Arg-1蛋白表达、M2型巨噬细胞比例明显升高（
P
<
0.05）。
结论
2
PTX3能够促进SAP小鼠肾组织细胞凋亡和肾损伤，其机制可能是通过诱导巨噬细胞向M1型分化。
Abstract
Objective
2
To explore the effect of PTX3 on kidney injury in mice with severe acute pancreatitis （SAP） and its possible mechanism.
Methods
2
Forty mice were randomly divided into the blank control group， the SAP group， the PTX3 group and the anti-PTX3 group， with 10 mice in each group. Except for the blank control group， mice in the other groups were injected with 20% L-arginine intraperitoneally to establish the SAP mouse model. Mice in the PTX3 group and the anti-PTX3 group were treated by intraperitoneal injection of recombinant mouse PTX3 （0.5 mg/kg） and PTX3 monoclonal antibody （0.2 mg/kg） respectively， while mice in the blank control group and the SAP group were injected with the same amount of normal saline， and all mice were killed after 24 hours. The levels of serum amylase （AMY）， creatinine （Cr） and blood urea nitrogen （BUN） were detected by automatic biochemical analyzer. The pathological changes of pancreas and renal tissue of mice were observed by HE staining. TUNEL staining was used to detect the apoptosis of renal tissue. The levels of serum interleukin （IL）-6， tumor necrosis factor-α （TNF-α）， IL-4 and IL-13 were detected by ELISA. The expression of PTX3， iNOS， CD16/32， CD206 and Arg-1 protein were detected by Western blot. The proportion of M1 and M2 type macrophages in renal tissue was detected by flow cytometry.
Results
2
Compared with the blank control group， the levels of AMY， BUN， Cr， IL-6， TNF-α， pathological score of pancreatic tissue， renal tubular injury score， apoptosis rate， the proportion of M1 type macrophages， the expression of PTX3， iNOS and CD16/32 protein in the SAP group were significantly increased （
P
<
0.05）， while the levels of IL-4 and IL-13， the expression of CD206 and Arg-1 protein， and the proportion of M2 type macrophages were significantly decreased （
P
<
0.05）. Compared with the SAP group， the levels of AMY， BUN， Cr， IL-6， TNF-α， pathological score of pancreatic tissue， renal tubular injury score， apoptosis rate， the proportion of M1 type macrophages， the expression of PTX3， iNOS and CD16/32 protein in the PTX3 group were significantly increased （
P
<
0.05）， while the levels of IL-4 and IL-13， the expression of CD206 and Arg-1 protein， and the proportion of M2 type macrophages were significantly decreased （
P
<
0.05）. Compared with the SAP group， the levels of AMY， BUN， Cr， IL-6， TNF-α， pathological score of pancreatic tissue， renal tubular injury score， apoptosis rate， the proportion of M1 type macrophages， the expression of PTX3， iNOS and CD16/32 protein in the anti-PTX3 group were significantly decreased （
P
<
0.05）， while the levels of IL-4 and IL-13， the expression of CD206 and Arg-1 protein， and the proportion of M2 type macrophages were significantly increased （
P
<
0.05）.
Conclusion
2
PTX3 can promote renal cell apoptosis and renal injury in SAP mice， possibly by inducing macrophages to differentiate into M1 type.
关键词
PTX3重症急性胰腺炎肾损伤机制
Keywords
PTX3severe acute pancreatitiskidney injurymechanism
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