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目的:探讨钙卫蛋白(CALP)在肾脏缺血再灌注损伤(IRI)大鼠中的表达及其在炎症反应中的作用。方法:50只雄性SD大鼠随机分为IRI组和假手术组(sham组),每组25只。各组分别于缺血再灌注后6 h、12h、24 h、48 h和72 h观察肾脏病理改变,检测大鼠血清尿素氮(BUN)和肌酐(SCr)水平,ELISA法检测大鼠血清CALP、肿瘤坏死因子(TNF)-α和白细胞介素(IL)-6的含量,免疫组织化学法和Western blot法分别检测大鼠肾脏CALP、Toll样受体4(TLR4)和NF-κB p65蛋白表达水平。结果:IRI组大鼠肾脏呈现不同程度的肾小管上皮细胞和间质损伤,伴炎细胞浸润。血清BUN、SCr、CALP及促炎细胞因子TNF-α、IL-6的含量在各时点均显著高于sham组(P<0.05)。CALP、TLR4和NF-κB p65主要表达于肾小管上皮细胞胞质中,在IRI组呈高表达,在各时点各因子的表达与sham组相比均显著增强(P<0.05)。结论:肾脏IRI大鼠的血清CALP、TNF-α和IL-6水平升高,CALP、TLR4和NF-κB p65在肾组织中表达增强。CALP在大鼠肾脏IRI的炎症反应中可能起重要作用。
Objective: To investigate the expression of calprotectin (CALP) in renal ischemia-reperfusion injury (IRI) rats and its role in the inflammatory response. Methods: Fifty male Sprague-Dawley rats were randomly divided into IRI group and sham operation group (sham group), 25 rats in each group. The pathological changes of kidney were observed at 6 h, 12 h, 24 h, 48 h and 72 h after ischemia-reperfusion. Serum urea nitrogen (BUN) and creatinine (SCr) were measured in all groups. Serum CALP , Tumor necrosis factor (TNF) -alpha and interleukin (IL) -6. Immunohistochemistry and Western blot were used to detect the expressions of CALP, Toll like receptor 4 (TLR4) and NF- The expression level. Results: The kidneys of IRI group showed different degrees of renal tubular epithelial cells and interstitial injury with infiltration of inflammatory cells. Serum levels of BUN, SCr, CALP and proinflammatory cytokines TNF-α and IL-6 were significantly higher than sham group at each time point (P <0.05). CALP, TLR4 and NF-κB p65 were mainly expressed in the cytoplasm of renal tubular epithelial cells, and were highly expressed in IRI group. The expression of CALP, TLR4 and NF-κB p65 in each group were significantly higher than those in sham group at each time point (P <0.05). CONCLUSIONS: Serum levels of CALP, TNF-α and IL-6 in renal IRI rats increased, while expression of CALP, TLR4 and NF-κB p65 in renal tissues increased. CALP may play an important role in the inflammatory reaction of rat kidney IRI.