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投稿时间：2023-04-12 网络发布日期：2023-11-20
投稿时间：2023-04-12 网络发布日期：2023-11-20
中文摘要: 目的 探讨干预P2X7受体对缺血再灌注（I/R）诱导肾脏纤维化的影响及可能的作用机制。方法 10只雄性野生型C57BL/6小鼠按随机数字表法分为假手术组（WT-Sham）、缺血再灌注损伤（IRI）组（WT-I/R）；10只以C57BL/6小鼠为背景的P2X7受体基因敲除小鼠同样随机分为假手术组(KO-Sham)、IRI伤组(KO-I/R)，每组各5只小鼠。采用一侧肾切除、对侧肾蒂夹闭法建立肾IRI模型。假手术组小鼠定位肾蒂但不进行肾切除和肾蒂夹闭。于造模或假手术42 d后收集小鼠肾组织标本。PAS染色观察肾组织结构改变，天狼猩红染色评估肾组织纤维化程度，Western blot法检测肾脏钙黏蛋白E（E-cadherin）、α-平滑肌肌动蛋白（α-SMA）的表达，免疫组化法观察肾脏F4/80阳性巨噬细胞浸润情况。结果 小鼠经过IRI 42 d后，PAS染色结果显示肾小管萎缩，间质可见大量炎症细胞浸润，天狼猩红染色提示肾组织明显纤维化。与野生型小鼠相比，P2X7受体基因敲除小鼠IRI 42d后肾小管萎缩程度减轻、炎症细胞浸润减少、肾脏纤维化情况改善。Western blot结果示，与假手术组比较，WT-I/R小鼠肾组织肾小管标记物E-cadherin表达降低，纤维化标记物α-SMA表达升高（P＜0.05）；与WT-I/R小鼠比较，KO-I/R小鼠E-cadherin降低程度和α-SMA升高程度均减轻（P＜0.05）。免疫组化结果示，小鼠肾组织经过IRI 42 d后巨噬细胞浸润增多，而敲除小鼠肾组织巨噬细胞浸润程度减轻。结论 IRI 可以导致肾组织巨噬细胞浸润和纤维化。干预P2X7受体可减少IRI 引起的巨噬细胞的浸润，改善肾组织纤维化，延缓慢性肾脏病进展。
Abstract:Objective To investigate the effects and possible mechanisms of intervening P2X7 receptors on ischemia/reperfusion (I/R)-induced renal fibrosis. Methods Ten male wild-type C57BL/6 mice were randomly assigned to the sham operation group (WT-Sham) and the renal ischemia-reperfusion injury(IRI) group (WT-I/R). Ten P2X7 receptors gene knockout mice were also randomly assigned to the sham operation group (KO-Sham) or the renal IRI group (KO-I/R), with five mice in each group. The renal IRI model was established by unilateral nephrectomy and contralateral renal pedicle clamping. In the sham operation group, mice underwent renal pedicle localization without nephrectomy or clamping. Renal tissue specimens were collected after 42 days of reperfusion or sham operation. PAS staining was used to observe changes in renal tissue structure, Picrosirius Red staining was used to assess the degree of renal fibrosis, Western blot was performed to detect the expression of renal calcium-adhering protein E-cadherin and α-smooth muscle actin （α-SMA）, and immunohistochemical staining was used to observe the infiltration of F4/80-positive macrophages in the kidneys. Results After 42 days of renal IRI, PAS staining showed tubular atrophy and significant infiltration of inflammatory cells in the interstitium, while Picrosirius Red staining indicated obvious renal fibrosis. Compared with wild-type mice, P2X7 receptors gene knockout mice showed reduced tubular atrophy, decreased infiltration of inflammatory cells, and improved renal fibrosis. Western blot results showed that compared to the sham operation group, WT-I/R mice had decreased expression of the tubular marker E-cadherin and increased expression of the fibrosis marker α-SMA (P＜0.05). Compared to WT-I/R mice, KO-I/R mice showed improvement in E-cadherin decrease and α-SMA increase (P＜0.05). Immunohistochemical results revealed increased infiltration of macrophages in the kidneys after 42 days of renal IRI, while P2X7 receptors gene knockout mice showed reduced macrophage infiltration. Conclusion Renal IRI can lead to macrophage infiltration and fibrosis in renal tissue. Intervention of P2X7 receptors can reduce macrophage infiltration caused by IRI, improve renal tissue fibrosis, and delay the progression of chronic kidney disease.
keywords: Chronic kidney disease Renal fibrosis P2X7 receptor Ischemia-reperfusion Ischemia-reperfusion injury Macrophage
文章编号： 中图分类号： 文献标志码：A
基金项目:国家自然科学基金项目（82000634，82100701）；浙江省自然科学基金项目（LQ21H050002）；浙江省医药卫生科技项目（2021427856）；上海市青年科技英才扬帆计划（20YF1425000）
附件
| Author Name | Affiliation |
| QIAN Yingying*, ZHAO Ning, WU Hongtao, WANG Ming, XIE Kewei | *Department of Nephrology, Hangzhou First People's Hospital, Hangzhou, Zhejiang 310006, China |
引用文本：
钱盈盈,赵宁,吴弘韬,等.干预P2X7受体对缺血再灌注所致肾脏纤维化的改善作用[J].中国临床研究,2023,36(11):1664-1667.
钱盈盈,赵宁,吴弘韬,等.干预P2X7受体对缺血再灌注所致肾脏纤维化的改善作用[J].中国临床研究,2023,36(11):1664-1667.