百里醌预处理对大鼠肾缺血再灌注损伤的影响

Influence of Thymoquinone pretreatment on renal ischemia-reperfusion injury in rats

摘要:

探讨百里醌对大鼠肾缺血再灌注损伤的影响及机制。方法随机将60 只雄性SD 大鼠分成假手术组（Sham 组）、肾缺血再灌注损伤组（IRI 组）和百里醌不同浓度预处理组（Thy 组）。IRI 组和Thy 组用无创血管夹夹闭左侧肾蒂复制IRI 模型，Sham 组操作同上，但不夹闭左侧肾蒂。Thy 组在缺血前45 min给予百里醌5、10、20和40 mg/kg腹腔注射。Sham组和IRI组在同一时间给予等体积生理盐水腹腔注射。再灌注24 h后处死小鼠，收集血清和肾脏标本。PAS 染色检测肾脏病理改变，ELISA 检测尿素氮（BUN）、肌酐（Cr）、丙二醛（MDA）、过氧化氢酶（CAT）、谷胱甘肽过氧化物酶（GPx）、超氧化物歧化酶（SOD）的表达、白细胞介素8（IL-8）、干扰素（IFN-γ）和肿瘤坏死因子（TNF-α）的表达,Western blot 检测JAK2、STAT3、P-JAK2、P-STAT3、P53 和P21 蛋白表达，逆转录聚合酶链反应（RT-PCR）检测IL-8、IFN-γ 和TNF-α 的信使RNA 水平（mRNA）。结果缺血再灌注损伤增加BUN、Cr、P-JAK2、P-STAT3、P53、P21、MDA、IL-8、IFN-γ和TNF-α 的表达以及肾组织病理改变，同时减少CAT、GPX 与SOD表达。而百里醌治疗，可减少BUN、Cr、P-JAK2、P-STAT3、P53、P21、MDA、IL-8、IFN-γ 和TNF-α表达以及肾组织病理改变，增加CAT、GPX 与SOD 表达，3 组之间JAK2 和STAT3 表达无差异。结论百里醌预处理可减轻肾缺血再灌注损伤，其作用机制部分是通过抑制JAK2/STAT3/P53 信号通路介导的炎症和氧化应激。

Abstract:

To explore the influence of Thymoquinone(Thy) on kidney ischemia-reperfusion injury (IRI)and the mechanism. Methods Sixty male Sprague-Dawley rats were randomly divided into sham operation (sham) group, IRI group and IR plus different concentration of Thy preconditioning groups. Thy groups were preconditioned with Thy (5, 10, 20 and 40 mg/kg) by intraperitoneal injection at 45 min prior to operation, and the other 2 groups were pretreated by equal volume of saline. The rats in the IRI and Thy groups were kept in 37℃infant incubators for 45 min after their left renal pedicles were clamped with noninvasive endoclips, and then their right kidneys were removed. The rats in the sham group underwent the same process, except for clamping the left renal pedicles. After 24-h reperfusion, blood samples and renal tissues were collected. The pathological changes of kidney tissues were observed using PAS staining. The expression levels of serum creatinine (Cr), urea nitrogen (BUN), IL-8, IFN-γ, TNF-α, MDA, CAT, GPX and SOD were measured by ELISA. The expression of JAK2, STAT3, p-JAK2, p-STAT3, p53 and p21 were measured by Western blot. Moreover, the mRNA levels of IL-8, IFN-γand TNF-αwere examined by RT-PCR. Results IRI markedly enhanced renal pathological changes and the expressions of BUN, Cr,p-JAK2, p-STAT3, p53, p21, MDA, IL-8, IFN-γand TNF-α, meanwhile reduced the expressions of CAT, GPX and SOD. Thymoquinone pretreatment significantly decreased renal pathological changes and the expressions ofBUN, Cr, p-JAK2, p-STAT3, p53, p21, MDA, IL-8, IFN-γand TNF-α, at the same time up-regulated the expressions of CAT, GPX and SOD. There was no significant difference in the expression of JAK2 or STAT3 among the three groups. Conclusions Thymoquinone pretreatment can alleviate kidney ischemia-reperfusion injury in rats partly by inhibiting inflammation and oxidative stress mediated by JAK2/STAT3/p53 signaling pathway.