摘要
目的:探究萝卜硫素通过Nrf2信号通路对大鼠草酸钙肾结石形成的作用和机制。方法:选取7~8周龄Wistar健康雄性大鼠30只,随机分为空白对照组、模型组和药物干预组。空白对照组给予正常饮用水,标准饲料;模型组给予1%乙二醇饮用水+2%氯化铵2 mL/d灌胃,标准饲料;药物干预组给予1%乙二醇饮用水+2%氯化铵2 mL/d灌胃+0.2 mg萝卜硫素腹腔注射,标准饲料。灌胃给药均连续28天。于第29天收分别集各组大鼠24 h尿液及肾脏标本,使用全自动生化分析仪检测尿液中K^(+)、Na^(+)、Cl^(+)、Ca^(2+)、Mg^(2+)、P^(5+)、Fe^(2+)含量,使用ELISA法测定尿液中丙二醛(malondialdehyde,MDA)与草酸水平,使用HE染色分析大鼠肾脏标本病理损伤情况,Von Kossa染色比较各组大鼠钙盐沉积情况,免疫组化法测定谷胱甘肽(glutathione,GSH)、核因子E_(2)相关因子2(nuclear factor erythroid-2 related factor 2,Nrf2)、8-羟基脱氧鸟苷(8-hydroxy-2 deoxyguanosine,8-OHDG)蛋白表达水平。结果:模型组大鼠尿液P^(5+)、Ca^(2+)、草酸、MDA含量显著高于空白对照组,HE染色示模型组肾小管较空白对照组变性严重,Von Kossa染色结果显示模型组钙盐沉积较空白对照组明显增多,模型组GSH、Nrf2免疫组化评分显著低于空白对照组,8-OHdG显著高于空白对照组,差异均有统计学意义(P<0.05);药物干预组大鼠尿液P^(5+)、Ca^(2+)、草酸、MDA含量显著低于模型组,HE染色、Von Kossa染色显示肾小管较模型组损伤减轻,钙盐沉积较模型组减轻,免疫组化显示GSH、Nrf2蛋白表达显著高于模型组,8-OHdG显著低于空白对照组,差异均有统计学意义(P<0.05)。结论:萝卜硫素对大鼠肾小管具有抗氧化损伤作用,可降低草酸钙晶体的释放,抑制草酸钙结石的形成,该作用可能是通过激活Nrf2蛋白及其下游信号通路实现的。
Objective:To investigate the effect of sulforaphane on the formation of calcium oxalate nephrolithiasis in rats through the nuclear factor E2-related factor 2(Nrf2)signaling pathway and its mechanism.Methods:Thirty healthy male Wistar rats aged 7-8weeks were selected and randomly divided into blank control group,model group and drug intervention group.The blank control group was given normal drinking water and standard feed.The model group was given drinking water containing 1%ethylene glycol+gavage with 2 mL/d 2%ammonium chloride and standard feed.The drug intervention group was given drinking water containing 1%ethylene glycol+gavage with 2 mL/d 2%ammonium chloride+intraperitoneal injection with 0.2 mg sulforaphane and standard feed.Gavage was treated for 28 d.On the 29th d,24 h urine and kidney specimens of rats in each group were collected,respectively.The contents of K^(+),Na^(+),Cl^(+),Ca^(2+),Mg^(2+),P^(5+)and Fe^(2+)in urine were detected by a full-automatic biochemical analyzer.The levels of malondialdehyde(MDA)and oxalic acid in urine were determined using ELISA.The pathological injury of rat kidney specimens was analyzed with HE staining.The calcium deposition of rats in each group was compared using Von Kossa staining.The protein expressions of glutathione(GSH),Nrf2 and 8-hydroxydeoxyguanosine(8-OHdG)were measured by immunohistochemistry.Results:The contents of P^(5+),Ca^(2+),oxalic acid and MDA in the urine of the model group were significantly higher than those in the blank control group.HE staining showed severer degeneration of the renal tubules in the model group compared with the blank control group.Von Kossa staining revealed that calcium salt deposition in the model group increased significantly than that in the blank control group.The immunohistochemical scores of GSH and Nrf2 in the model group were significantly lower than those in the blank control group,while 8-OHdG was significantly higher than that in the blank control group(P<0.05).In the drug intervention group,the contents of urine P^(5+),Ca^(2+),oxalic acid and MDA were significantly lower than those in the model group.HE staining and Von Kossa staining presented that the injury of the renal tubules and calcium salt deposition in the drug intervention group were milder than those in the model group.Immunohistochemistry demonstrated that the protein expressions of GSH and Nrf2 were significantly higher than those in the model group,while 8-OHdG was significantly lower than that in the blank control group(P<0.05).Conclusion:Sulforaphane has a protective effect on oxidant damage to rat renal tubules,as well as can reduce the release of calcium oxalate crystals and inhibit the formation of calcium oxalate stones,which may be achieved by activating Nrf2 protein and its downstream signaling pathway.
作者
刘若天
木拉提·马合木提
郜乐
杨奇
陈策
拜合提亚尔·艾合买提江
麦吾拉江·买合木提
LIU Ruo-tian;MULATI·Mahemuti;GAO Le;YANG Qi;CHEN Ce;BAIHETIYAER·Aihemaitijiang;MAIWULAJIANG·Maihemuti(The Second Affiliated Hospital of Xinjiang Medical University,Urumqi,Xinjiang,830000,China)
出处
《现代生物医学进展》
CAS
2022年第11期2028-2033,共6页
Progress in Modern Biomedicine
基金
新疆维吾尔自治区自然科学基金项目(2017D01C254)。
