摘要
目的 通过研究肾缺血再灌注损伤时肾组织中胆绿素还原酶(biliverdin reductase, BVR)的表达与巨噬细胞的关系,为进一步研究BVR与巨噬细胞在肾脏纤维化中的作用提供依据。方法 16只C57BL/6雄性小鼠随机分为假手术组(Sham组)和肾脏缺血再灌注组(ischemia-reperfusion injury,IRI组),取肾组织切片行PAS、Masson、Sirius red染色,免疫组织化学检测α-SMA、PDGF-β和Collagen IV的表达,RT-PCR检测α-SMA、FN、Collagen I和KIM-1 mRNA表达,流式细胞术和免疫荧光染色检测BVR与巨噬细胞的关系。结果 在缺血再灌注小鼠损伤肾组织内,PAS染色发现IRI组肾小球硬化明显,系膜细胞增生,肾小管管腔扩张,小管上皮细胞肿胀或发生空泡变性、细胞脱落、基膜裸露。Masson和Sirius Red染色发现肾间质胶原纤维表达明显增加;免疫组织化学染色显示IRI组α-SMA、PDGF-β和collagen IV免疫反应性增强,RTPCR检测显示IRI组α-SMA、FN、collagen I和KIM-1 mRNA水平增高;流式细胞术检测发现,在肾缺血再灌注小鼠,血液、骨髓和肾组织BVR阳性巨噬细胞显著增加,免疫荧光双标进一步证实缺血再灌注肾损伤组织BVR阳性巨噬细胞明显增多。结论 BVR阳性巨噬细胞参与肾缺血再灌注损伤后肾纤维化过程。
Objective By studying the relationship between the expression of biliverdin reductase(BVR) in renal tissue and macrophages during renal ischemia-reperfusion injury, to provide a basis for further study on the role of BVR and macrophages in renal fibrosis. Methods Sixteen C57BL/6 male mice were randomly divided into two groups: sham operation group(Sham group,n = 8), renal ischemia-reperfusion injury group(IRI group, n = 8). Kidney tissues were used for HE, Masson and Sirius red stains to assess histological changes, immunohistochemical detection of α-SMA, PDGF-β, Collagen I expression in ischemia-reperfusion injury group and sham operation group, RT-PCR method for α-SMA, FN, Collagen I, KIM-1expression in IRI group and Sham group, flow cytometry and immunofluorescence to detect the relationship between BVR and macrophages. Results Compared with the sham-operated group, in the ischemia-reperfusion model, renal interstitial fibrosis and inflammatory cell infiltration were significantly worse.Immunohistochemistry showed increased expression of α-SMA, PDGF-β and collagen IV in the IRI group. RT-PCR showed increased expression of α-SMA, FN, Collagen I, and KIM-1 in the IRI group. In mice with renal ischemia-reperfusion injury, flow cytometry analysis found that the number of BVR positive macrophages in blood, bone marrow and kidney tissue increased significantly, and the double immunofluorescence staining further confirmed the increase of BVR positive macrophages in the renal tissue. Conclusion BVR positive macrophages might participate in the process of renal fibrosis after renal ischemia-reperfusion injury.
作者
胡芝芝
裴广畅
曾锐
徐钢
Hu Zhizhi;Pei Guangchang;Zeng Rui;Xu Guang(Division of nephrology,Department of internal medicine,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China)
出处
《中国组织化学与细胞化学杂志》
CAS
CSCD
2022年第6期551-557,574,共8页
Chinese Journal of Histochemistry and Cytochemistry
基金
国家自然科学基金青年项目(81700653)。
关键词
肾缺血再灌注损伤
胆绿素还原酶
巨噬细胞
肾纤维化
Renal ischemia-reperfusion injury
biliverdin reductase
macrophage
renal fibrosis
