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目的观察大鼠缺血再灌注损伤模型中,瞬时受体电位通道M7(trallsient receptor potential melastatiIl related7,TRPM7)的表达变化及其与肾组织病理损伤、程序性坏死标志物RIP1、聚腺苷酸二磷酸核糖转移酶-1(PARP-1)表达变化的关系,初步探讨TRPM7在肾脏缺血再灌注损伤细胞程序性坏死中的作用及其可能的机制。方法 55只雄性SD大鼠建立缺血再灌注模型后,于再灌注后24、48 h、5、7 d取动脉血检测大鼠血肌酐、尿素氮水平;肾组织标本HE染色观察肾组织病理损伤;免疫组化和Western blot检测肾组织TRPM7、PARP-1、RIP1定位及表达变化。结果对照组和假手术组相比,血肌酐尿素氮水平、肾脏组织形态无明显变化;与对照组、假手术组相比,缺血再灌注后大鼠血肌酐尿素氮水平明显升高,再灌注后24 h达到最高值,随后逐渐下降,7 d左右降至接近正常。HE染色显示肾组织在再灌注24、48 h均出现明显急性肾小管损伤,5 d时开始修复,至第7天基本修复。免疫组化及Western blot显示,TRPM7主要在肾小管上皮细胞表达,再灌注后TRPM7表达显著升高,其中24 h达高峰,随后逐渐下降。PARP-1、RIP1的表达变化有相似的趋势。相关性分析显示TRPM7表达变化与肾小管急性损伤评分呈正相关(r=0.71,P<0.05)。结论瞬时受体电位通道M7可能参与了肾脏缺血再灌注损伤,通过下游钙依赖的信号传导途径调控PARP-1、RIP1影响程序性坏死发挥作用。
Objective To investigate the expression of TRPM7 in rat model of ischemia-reperfusion injury and its correlation with pathological changes of renal tissue, markers of programmed necrosis RIP1, poly (A) (PARP-1), and to explore the role of TRPM7 in programmed cell death induced by renal ischemia / reperfusion injury and its possible mechanism. Methods Fifty-five male Sprague-Dawley rats were used to establish the model of ischemia-reperfusion. Blood samples were collected at 24, 48, 54 and 72 hours after reperfusion for the determination of serum creatinine and blood urea nitrogen. The renal pathological changes The expression of TRPM7, PARP-1, RIP1 in renal tissue was detected by immunohistochemistry and Western blot. Results Compared with the sham-operation group, the levels of serum urea nitrogen and serum creatinine did not change significantly in the control group and the sham operation group. Compared with the control group and the sham-operated group, the serum creatinine and urea nitrogen level of the rats after ischemia-reperfusion were significantly increased 24 h after perfusion reached the highest value, then gradually decreased, around 7 d reduced to near normal. Hematoxylin and eosin staining revealed that renal tissue showed obvious acute tubular injury at 24 h and 48 h after reperfusion, repaired on day 5, and repaired on day 7. Immunohistochemistry and Western blot showed that TRPM7 was mainly expressed in renal tubular epithelial cells. After reperfusion, the expression of TRPM7 increased significantly, reaching its peak at 24 h and then gradually decreased. A similar trend was observed in the expression of PARP-1 and RIP1. Correlation analysis showed that there was a positive correlation between TRPM7 expression and acute tubular injury score (r = 0.71, P <0.05). Conclusions M7, a transient receptor potential pathway, may be involved in renal ischemia-reperfusion injury. PARP-1 may be regulated by the downstream calcium-dependent signaling pathway. RIP1 may play a role in programmed necrosis.