辣椒素减轻大鼠肾缺血-再灌注损伤的作用
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摘要
目的探讨辣椒素减轻大鼠肾缺血-再灌注损伤(ischemia-reperfusion injury,IRI)的作用及其机制。方法成年健康清洁雄性SD大鼠40只,体重200～250 g,随机分为四组:假手术组(S组)、缺血-再灌注组(IR组)、辣椒素组(CAP组)和辣椒素+辣椒平组(CPZ组),每组10只。S组仅分离双侧肾蒂不结扎,IR组、CAP组和CPZ组通过动脉夹夹闭双侧肾蒂45 min,再灌注24 h的方法建立肾缺血-再灌注模型。S组与IR组在同时点注射等容量生理盐水,CAP组和CPZ组大鼠于缺血前30 min经腹腔注射辣椒素30 mg/kg。CPZ组大鼠于缺血前45 min经腹腔注射辣椒平20 mg/kg。于再灌注24 h后取血标本检测大鼠血清肌酐(Cr)、尿素氮(BUN)、白细胞介素-6(IL-6),肿瘤坏死因子-α(TNF-α)和白细胞介素-10(IL-10)浓度。采用Western blot法检测Janus蛋白酪氨酸激酶2(JAK2)、信号转导子及转录激活子3(STAT3)、磷酸化Janus蛋白酪氨酸激酶2(P-JAK2)、磷酸化信号转导子及转录激活子3(P-STAT3)和caspase-3的蛋白含量。取肾组织观察组织病理学改变,检测凋亡率(AI)。结果与S组比较,IR组、CAP组和CPZ组血清Cr、BUN、IL-6、TNF-α、IL-10浓度、P-JAK2/JAK2、P-STAT3/STAT3和caspase-3蛋白含量明显升高(P<0.05),肾小管上皮细胞出现不同程度的肿胀,空泡变性,刷状缘脱落,部分肾小管管腔扩张,肾小管损伤评分和AI明显升高(P<0.05);与IR组和CPZ组比较,CAP组血清Cr、BUN、IL-6、TNF-α浓度、P-JAK2/JAK2、P-STAT3/STAT3和caspase-3含量明显降低(P<0.05),肾小管损伤减轻,肾小管损伤评分和AI明显降低(P<0.05),IL-10浓度明显升高(P<0.05)。结论辣椒素可减轻缺血-再灌注引起的大鼠肾组织损伤,其机制可能与抑制IL-6/JAK2/STAT3通路,抑制炎症反应,减少细胞凋亡和组织损伤有关。
Objective To investigate the effect and mechanism of capsaicin on renal ischemia-reperfusion injury in rats. Methods Forty healthy male SD rats weighing 200-250 g were randomly divided into 4 groups(n= 10): sham operation group(group S), ischemia-reperfusion group(group IR),capsaicin treatment group(group CAP) and both capsazepine and capsaicin treatment group(group CPZ). The renal ischemia-reperfusion injury model was established by clamping bilateral renal pedicle for 45 min and reperfusing for 24 h. Rats in the group CAP and group CPZ were administrated with capsaicin 30 mg/kg via intraperitoneal injection 30 min before renal ischemia. Rats in group CPZ were treated with capsazepine 20 mg/kg 15 min prior to administration of capsaicin via intraperitoneal injection.Rats in group S and group IR were administrated with saline. At 24 h after reperfusion, the blood sample was obtained to detect the level of serum concentration of creatinine(Cr), blood urea nitrogen(BUN), tumor necrosis factor-α(TNF-α),interleukin-6(IL-6) and interleukin-10(IL-10).Renal tissue was obtained to observe the pathological changes(via HE stain), the apoptotic rate(by TUNEL) and expression of Janus kinase 2(JAK2), signal transducer and activator of transcription 3(STAT3), phosphorylated Janus kinase 2(P-JAK2), phosphorylated signal transducer and activator of transcription 3(P-STAT3) and caspase-3(with Western blotting). Results Compared with group S, the levels of Cr, BUN, IL-6, TNF-α, IL-10 and the expression of P-JAK2/JAK2, P-STAT3/STAT3, caspase-3 were increased significantly. Besides, tubule epithelial cells showed drfferent degrees of swelling, vacuolow degeneration, brush margin shelding, part of the tubular cavity diluted, renal tubular damage score and AI were significantly increased in group IR(P < 0.05). Compared with group IR and CPZ, the levels of Cr, BUN, IL-6, TNF-α and the expression of P-JAK2/JAK2, P-STAT3/STAT3, caspase-3 decreased significantly(P < 0.05). Renal tubule inijury allevicated significantly. Renal tubular damage score and AI in group CAP decreased significantly(P < 0.05) and IL-10 increased significantly(P < 0.05). Conclusion Administration of capsaicin alleviated renal IRI which might attenuate inflammatory responses via IL-6/JAK2/STAT3 signaling pathway.
Objective To investigate the effect and mechanism of capsaicin on renal ischemia-reperfusion injury in rats. Methods Forty healthy male SD rats weighing 200-250 g were randomly divided into 4 groups(n= 10): sham operation group(group S), ischemia-reperfusion group(group IR),capsaicin treatment group(group CAP) and both capsazepine and capsaicin treatment group(group CPZ). The renal ischemia-reperfusion injury model was established by clamping bilateral renal pedicle for 45 min and reperfusing for 24 h. Rats in the group CAP and group CPZ were administrated with capsaicin 30 mg/kg via intraperitoneal injection 30 min before renal ischemia. Rats in group CPZ were treated with capsazepine 20 mg/kg 15 min prior to administration of capsaicin via intraperitoneal injection.Rats in group S and group IR were administrated with saline. At 24 h after reperfusion, the blood sample was obtained to detect the level of serum concentration of creatinine(Cr), blood urea nitrogen(BUN), tumor necrosis factor-α(TNF-α),interleukin-6(IL-6) and interleukin-10(IL-10).Renal tissue was obtained to observe the pathological changes(via HE stain), the apoptotic rate(by TUNEL) and expression of Janus kinase 2(JAK2), signal transducer and activator of transcription 3(STAT3), phosphorylated Janus kinase 2(P-JAK2), phosphorylated signal transducer and activator of transcription 3(P-STAT3) and caspase-3(with Western blotting). Results Compared with group S, the levels of Cr, BUN, IL-6, TNF-α, IL-10 and the expression of P-JAK2/JAK2, P-STAT3/STAT3, caspase-3 were increased significantly. Besides, tubule epithelial cells showed drfferent degrees of swelling, vacuolow degeneration, brush margin shelding, part of the tubular cavity diluted, renal tubular damage score and AI were significantly increased in group IR(P < 0.05). Compared with group IR and CPZ, the levels of Cr, BUN, IL-6, TNF-α and the expression of P-JAK2/JAK2, P-STAT3/STAT3, caspase-3 decreased significantly(P < 0.05). Renal tubule inijury allevicated significantly. Renal tubular damage score and AI in group CAP decreased significantly(P < 0.05) and IL-10 increased significantly(P < 0.05). Conclusion Administration of capsaicin alleviated renal IRI which might attenuate inflammatory responses via IL-6/JAK2/STAT3 signaling pathway.
引文
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[8] Correa-Costa M,Azevedo H,Amano MT,et al.Transcriptome analysis of renal ischemia/reperfusion injury and its modulation by ischemic pre-conditioning or hemin treatment.PLoS One,2012,7(11):e49569.
[9] 斯妍娜,鲍红光,张勇,等.右美托咪定对大鼠肾缺血-再灌注损伤的保护作用.临床麻醉学杂志,2013,29(3):261-264.
[10] Zhao H,Alam A,Soo AP,et al.Ischemia-reperfusion injury reduces long term renal graft survival:mechanism and beyond.EBioMedicine,2018,28:31-42.
[11] Bonventre JV,Yang L.Cellular pathophysiology of ischemic acute kidney injury.J Clin Invest,2011,121(11):4210-4221.
[12] Brosius FC,Tuttle KR,Kretzler M.JAK inhibition in the treatment of diabetic kidney disease.Diabetologia,2016,59(8):1624-1627.
[13] Lu TC,Wang ZH,Feng X,et al.Knockdown of Stat3 activity in vivo prevents diabetic glomerulopathy.Kidney Int,2009,76(1):63-71.
[2] Kusch A,Hoff U,Bubalo G,et al.Novel signalling mechanisms and targets in renal ischaemia and reperfusion injury.Acta physiol(Oxf),2013,208(1):25-40.
[3] 张炯,王佳,王芳,等.辣椒素减轻肾缺血再灌注损伤的线粒体相关机制.中国动脉硬化杂志,2018,26(6):583-587.
[4] Huang M,Cheng G,Tan H,et al.Capsaicin protects cortical neurons against ischemia/reperfusion injury via down-regulating NMDA receptors.Exp Neurol,2017,295:66-76.
[5] Gao Y,Song J,Chen H,et al.TRPV1 activation is involved in the cardioprotection of remote limb ischemic postconditioning in ischemia-reperfusion injury rats.Biochem Biophys Res Commun,2015,463 (4):1034-1039.
[6] Ueda K,Tsuji F,Hirata T,et al.Preventive effect of TRPV1 agonists capsaicin and resiniferatoxin on ischemia/reperfusion-induced renal injury in rats.J Cardiovasc Pharmacol,2008,51(5):513-520.
[7] Yang N,Luo M,Li R,et al.Blockage of JAK/STAT signalling attenuates renal ischaemia-reperfusion injury in rats.Nephrol Dial Transplant,2008,23(1):91-100.
[8] Correa-Costa M,Azevedo H,Amano MT,et al.Transcriptome analysis of renal ischemia/reperfusion injury and its modulation by ischemic pre-conditioning or hemin treatment.PLoS One,2012,7(11):e49569.
[9] 斯妍娜,鲍红光,张勇,等.右美托咪定对大鼠肾缺血-再灌注损伤的保护作用.临床麻醉学杂志,2013,29(3):261-264.
[10] Zhao H,Alam A,Soo AP,et al.Ischemia-reperfusion injury reduces long term renal graft survival:mechanism and beyond.EBioMedicine,2018,28:31-42.
[11] Bonventre JV,Yang L.Cellular pathophysiology of ischemic acute kidney injury.J Clin Invest,2011,121(11):4210-4221.
[12] Brosius FC,Tuttle KR,Kretzler M.JAK inhibition in the treatment of diabetic kidney disease.Diabetologia,2016,59(8):1624-1627.
[13] Lu TC,Wang ZH,Feng X,et al.Knockdown of Stat3 activity in vivo prevents diabetic glomerulopathy.Kidney Int,2009,76(1):63-71.