中国组织工程研究 ›› 2019, Vol. 23 ›› Issue (23): 3643-3648.doi: 10.3969/j.issn.2095-4344.1309

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

姜黄素后处理肢体缺血再灌注肾损伤模型大鼠基质金属蛋白酶9及基质金属蛋白酶组织抑制因子1平衡的变化

邹海波,孙晓峰   

  1. (沈阳医学院附属中心医院麻醉科,辽宁省沈阳市  110024)
  • 收稿日期:2019-02-18 出版日期:2019-08-18 发布日期:2019-08-18
  • 通讯作者: 孙晓峰,博士,主任医师,沈阳医学院附属中心医院麻醉科,辽宁省沈阳市 110024
  • 作者简介:邹海波,男,1981年生,辽宁省沈阳市人,2008年中国医科大学毕业,硕士,副主任医师,主要从事临床危重症患者麻醉的器官保护性研究。
  • 基金资助:

    辽宁省教育厅一般项目(L2015540),项目负责人:孙晓峰;沈阳医学院青年基金项目(20162029),项目负责人:邹海波

Effects of curcumin post-conditioning on the balance of matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 in a rat model of renal injury induced by limb ischemia/reperfusion

Zou Haibo, Sun Xiaofeng   

  1.  (Department of Anesthesiology, Affiliated Central Hospital of Shenyang Medical College, Shenyang 110024, Liaoning Province, China)
  • Received:2019-02-18 Online:2019-08-18 Published:2019-08-18
  • Contact: Sun Xiaofeng, MD, Chief physician, Department of Anesthesiology, Affiliated Central Hospital of Shenyang Medical College, Shenyang 110024, Liaoning Province, China
  • About author:Zou Haibo, Master, Associate chief physician, Department of Anesthesiology, Affiliated Central Hospital of Shenyang Medical College, Shenyang 110024, Liaoning Province, China
  • Supported by:

    the General Project of Education Department of Liaoning Province, No. L2015540 (to SXF); the Youth Foundation of Shenyang Medical College, No. 20162029 (to ZHB)

摘要:

文章快速阅读:

文题释义:

姜黄素:是从姜科、南星科中的一些植物的根茎中提取的一种化学成分,具有抗氧化应激、抑制炎症细胞因子释放、抗细胞凋亡等广泛的药理活性。
基质金属蛋白酶9/基质金属蛋白酶组织抑制因子1:基质金属蛋白酶9在肢体缺血再灌注致肾损伤的病理发展过程中发挥着重要作用,而基质金属蛋白酶组织抑制因子1是基质金属蛋白酶9的抑制物,基质金属蛋白酶9/基质金属蛋白酶组织抑制因子1比值的平衡对维持肾脏组织正常生理功能和结构稳定起极其重要的作用。
摘要
背景:
姜黄素预处理可减轻肢体缺血再灌注对肾脏的损伤,但姜黄素后处理是否对肾脏损伤有保护作用及其机制目前研究甚少。
目的:探讨大鼠肢体缺血再灌注肾组织损伤时姜黄素后处理对基质金属蛋白酶9/基质金属蛋白酶组织抑制因子1平衡的影响。
方法:成年雄性SD大鼠80只由中国医科大学实验室中心提供。实验方案经沈阳医学院附属中心医院动物实验伦理委员会批准。采用随机数字表法,将80只SD大鼠分为4组(n=20):假手术组、缺血再灌注组、姜黄素后处理组和地塞米松组。除假手术组外,其余大鼠采用夹闭双侧股动脉4 h,再灌注4 h的方法制备肢体缺血再灌注模型。在大鼠肢体缺血后4 h,姜黄素后处理组经腹腔注射姜黄素200 mg/kg,地塞米松组经腹腔注射地塞米松1.5 mg/kg,假手术组、缺血再灌注组以等量生理盐水替代。于再灌注4 h后,经颈动脉采集血样,检测肌酐、尿素氮;麻醉下取大鼠肾脏组织检测丙二醛水平,苏木精-伊红染色检测病理变化和肾小管间质半定量评分(SSRT);应用Westen blot法和RT-PCR法分别测定基质金属蛋白酶9、基质金属蛋白酶组织抑制因子1蛋白和核因子κB p65 mRNA含量,计算基质金属蛋白酶9/基质金属蛋白酶组织抑制因子1比值;应用ELESA法检测肺组织肿瘤坏死因子α和白细胞介素1β等炎性因子的水平。
结果与结论:①与假手术组比较,缺血再灌注组血浆肌酐、尿素氮和丙二醛均升高(P < 0.05);肾苏木精-伊红染色示肾小管扩张,管腔内可见管型,刷状缘消失、肾间质炎症细胞浸润等病变; SSRT升高(P < 0.05);基质金属蛋白酶9、基质金属蛋白酶9/基质金属蛋白酶组织抑制因子1比值和核因子κB p65 mRNA含量均升高(P < 0.05);肿瘤坏死因子α和白细胞介素1β炎性因子释放增加(P < 0.05);②与缺血再灌注组比较,姜黄素后处理组血浆肌酐、尿素氮和丙二醛降低(P < 0.05);肾组织炎性细胞浸润数量减少,管型消失,偶可见肾小管扩张;SSRT降低(P < 0.05);基质金属蛋白酶9蛋白、基质金属蛋白酶9/基质金属蛋白酶组织抑制因子1比值和NF-κB p65 mRNA含量降低(P < 0.05);肿瘤坏死因子α和白细胞介素1β炎性因子释放降低(P < 0.05);③与姜黄素后处理组与地塞米松组比较各指标差异无显著性意义;④结果表明,姜黄素后处理可减轻大鼠肢体缺血再灌注造成的肾损伤,其机制与降低组织基质金属蛋白酶9/基质金属蛋白酶组织抑制因子1比值有关。

关键词: 姜黄素, 缺血再灌注, 姜黄素后处理, 肾, 基质金属蛋白酶9, 基质金属蛋白组织抑制因子1, 肢体缺血再灌注, 肾损伤模型

Abstract:

BACKGROUND: Curcumin preconditioning can alleviate renal injury induced by limb ischemia/reperfusion, but whether curcumin preconditioning has protective effect on renal injury and its mechanisms are still poorly studied.
OBJECTIVE: To investigate the effects of the balance of matrix metalloproteinase-9/tissue inhibitor of metalloproteinase-1 when curcumin post-conditioning towards renal injury induced by limb ischemia/reperfusion in rats.
METHODS: Eighty adult male Sprague-Dawley rats were provided by Laboratory Center of China Medical University, and the study was approved by the Laboratory Animal Ethics Committee of Affiliated Central Hospital of Shenyang Medical College. The rats were randomly divided into four groups (n=20/group) using a random number table: sham, ischemia/reperfusion, curcumin post-conditioning and dexamethasone groups. Limb ischemia was induced by occlusion of bilateral femoral arteries of rats except for sham group for 4 hours, followed by reperfusion for 4 hours. At 4 hours after ischemia, 200 mg/kg curcumin and 1.5 mg/kg dexamethasone was immediately injected by intraperitoneal pathway into curcumin post-conditioning and dexamethasone groups, respectively. The same volume of normal saline was injected into the rats in the other two groups. Arteries blood samples were taken at 4 hours of reperfusion for examining biochemical indexes such as creatinine and blood urea nitrogen. Kidney was removed to detect the level of malondialdehyde. Hematoxylin-eosin staining was used to detect pathological changes and semi-quantitative score of renal tubulointerstitia. The expression levels of matrix metalloproteinase-9, tissue inhibitor of metalloproteinase-1 and nuclear factor-κB p65 mRNA were detected by western blot assay and RT-PCR, respectively. The levels of tumor necrosis factor-α and interleukin-1β in lung tissue were detected by ELISA.
RESULTS AND CONCLUSION: (1) Compared with sham group, the levels of creatinine, blood urea nitrogen and malondialdehyde in the ischemia/reperfusion group were significantly increased (P < 0.05). Hematoxylin-eosin staining showed obvious dilatation of renal tubules, tubular pattern, and disappearance of brush border and infiltration of inflammatory cells in renal interstitium in the lesion field. Semi-quantitative score of renal tubulointerstitia increased (P < 0.05). The expression levels of matrix metalloproteinase-9, nuclear factor-κB p65 mRNA and matrix metalloproteinase-9/ tissue inhibitor of metalloproteinase-1 were increased (P < 0.05). The release of tumor necrosis factor and interleukin-1 was increased (P < 0.05). (2) Compared with the ischemia/reperfusion group, the levels of creatinine, blood urea nitrogen and malondialdehyde in the curcumin post-conditioning group were significantly decreased (P < 0.05). Hematoxylin-eosin staining showed obvious dilatation of renal tubules, tubular pattern, and disappearance of brush border and infiltration of inflammatory cells in renal interstitium in the lesion field. Semi-quantitative score of renal tubulointerstitia decreased (P < 0.05). The expression levels of matrix metalloproteinase-9, nuclear factor-κB p65 mRNA and matrix metalloproteinase-9/ tissue inhibitor of metalloproteinase-1 were decreased (P < 0.05). The release of tumor necrosis factor and interleukin-1 was decreased (P < 0.05). (3) There was no significant difference in each index between curcumin post-conditioning and dexamethasone groups. (4) In summary, curcumin post-treatment can reduce the renal injury induced by limb ischemia/reperfusion injury in rats, which may be related to the reduction of matrix metalloproteinase-9/tissue inhibitor of metalloproteinase-1.

Key words:  curcumin, ischemia/reperfusion, curcumin post-conditioning, renal, matrix metalloproteinase 9, tissue inhibitor of metalloproteinase-1, limb ischemia/reperfusion, renal injury model

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