萝卜硫素对猪复苏后肾损伤影响

Effect of Sulforaphane on Postresuscitati on Renal Injury in Pigs

目的探讨萝卜硫素(SFN)对猪复苏后肾损伤的影响及作用机制。方法将22头国产大白猪随机分为假手术组(Sham,n=6)、心肺复苏组(CPR,n=8)和CPR+SFN组(n=8)。利用心脏骤停10 min与心肺复苏6 min建立动物模型。于复苏后5 min时,CPR+SFN组经股静脉泵入2 mg/kg SFN。于复苏后1、2、4、24 h时,采集血标本检测肌酐(SCr)和尿素氮(BUN)的血清浓度。于复苏后24 h时取左肾上极组织标本,检测肾组织的铁沉积水平,丙二醛(MDA)和还原性谷胱甘肽(GSH)含量,超氧化物歧化酶(SOD)活性;Western Blot法检测核因子E2相关因子2(Nrf2)、血红素加氧酶1(HO-1)和谷胱甘肽过氧化物酶4(GPX4)蛋白表达水平。结果与Sham组比较,CPR组SCr和BUN血清浓度在复苏后24 h观察期间的各时间点均升高,肾组织铁沉积水平和MDA含量升高,GSH含量和SOD活性降低,Nrf2和HO-1蛋白表达增加,GPX4蛋白表达减少(均P<0.05)。与CPR组比较,CPR+SFN组SCr和BUN血清浓度在复苏2 h后降低;在复苏后24 h时的肾组织铁沉积水平和MDA含量降低,GSH含量和SOD活性升高,Nrf2、HO-1和GPX4蛋白表达增加(均P<0.05)。结论SFN能减轻猪复苏后肾损伤,其保护机制可能与激活Nrf2/HO-1/GPX4通路后抑制细胞铁死亡的过程有关。

Objective To study the effect and mechanism of sulforaphane(SFN)on postresuscitation renal injury in pigs.Methods Twenty-two domestic white pigs were randomly divided into the Sham group(n=6),cardiopulmonary resuscitation group(CPR,n=8),and CPR+SFN group(n=8).The animal model was established by using the setting of 10 min of cardiac arrest and then 6 min of CPR.At 5 min after resuscitation,a dose of 2 mg/kg of SFN was pumped into pigs of the CPR+SFN group through femoral veins.At respectively 1,2,4,and 24 h after resuscitation,blood samples were collected for the measurement of concentrations of serum creatinine(SCr)and blood urea nitrogen(BUN).At 24 h after resuscitation,the left renal upper pole tissue specimens was taken to measure the iron deposition level,malondialdehyde(MDA)and glutathione(GSH)contents and the superoxide dismutase(SOD)activity in kidneys.Western Blot was used to test the protein expression levels of nuclear factor E2-related factor 2(Nrf2),heme oxygenase 1(HO-1),and glutathione peroxidase 4(GPX4).Results Compared with the Sham group,the CPR group showed increased serum concentrations of SCr and BUN at all time points during 24 h of post-resuscitation observation,and raised iron deposition level and MDA content in kidneys.The GSH contents and SOD activity reduced,the protein expression levels of Nrf2 and HO-1 increased,while the protein expression of GPX4 decreased(all P<0.05).Compared with CPR group,the serum concentrations of SCr and BUN decreased at 2 h after resuscitation in the CPR+SFN group.At 24 h after resuscitation,the iron deposition level and the MDA content in kidneys reduced,while the GSH content and SOD activity as well as the protein expression levels of Nrf2,HO-1,and GPX4 increased(all P<0.05).Conclusions SFN could alleviate renal injury in pigs after resuscitation.Its mechanism might be associated with the inhibition process of cell ferroptosis through activating the Nrf2/HO-1/GPX4 pathway.