摘要
目的探讨高压电烧伤后大鼠肾脏氧化应激损伤的特点及灯盏花素的干预作用。方法该研究为实验研究。将160只8~10周龄雄性SD大鼠按随机数字表法分为假伤组、电烧伤组、盐水组、低灯盏花素组、中灯盏花素组、高灯盏花素组,其中假伤组、电烧伤组各60只大鼠,余4组各10只大鼠。将假伤组和电烧伤组大鼠按伤后0 h(即刻)、8 h、24 h、48 h、72 h和1周,分为每个时间点10只。将假伤组大鼠不通电致假伤,余5组大鼠致高压电烧伤。伤后对假伤组和电烧伤组大鼠不行干预,对盐水组、低灯盏花素组、中灯盏花素组、高灯盏花素组大鼠分别另经腹腔按5 mL/kg剂量注射生理盐水及0.4、1.6、4.0 g/L灯盏花素,每24小时1次,至伤后72 h。模型制作成功后14只大鼠死亡,包括伤后24 h、48 h、72 h、1周电烧伤组各1、2、2、1只,伤后72 h盐水组、低灯盏花素组、中灯盏花素组、高灯盏花素组各4、1、2、1只。取6组大鼠的肾脏组织,称重后计算肾/体比。取假伤组,伤后8 h、24 h、48 h、72 h和1周的电烧伤组,伤后72 h的盐水组、低灯盏花素组、中灯盏花素组、高灯盏花素组各4只大鼠的左肾上极组织,行苏木精-伊红染色后采用半定量病理评分系统行肾小管与肾间质损伤评分。取6组大鼠下腔静脉血,采用肌氨酸氧化酶法检测血清肌酐水平,采用脲酶法检测血清尿素氮水平;取6组大鼠右肾皮质,采用钼酸铵法检测肾组织上清液中过氧化氢酶(CAT)活力,采用酶联免疫吸附测定法检测肾组织上清液中晚期氧化蛋白产物(AOPP)水平和Klotho蛋白水平。结果伤后8 h、48 h、72 h、1周,电烧伤组大鼠肾/体比均明显高于假伤组(t值分别为-0.52、-3.75、-4.05、-2.25,P<0.05)。伤后72 h,与电烧伤组、盐水组、低灯盏花素组、中灯盏花素组比较,高灯盏花素组大鼠肾/体比明显降低(P值均<0.05)。与假伤组比较,电烧伤组大鼠伤后48 h、72 h、1周肾小管与肾间质损伤评分均明显升高(P<0.05);与电烧伤组伤后8、24 h比较,电烧伤组大鼠伤后1周肾小管与肾间质损伤评分明显升高(P值均<0.05)。伤后72 h,5组电烧伤大鼠肾小管与肾间质损伤评分相近(P>0.05)。伤后8 h、24 h、48 h、72 h、1周,电烧伤组大鼠血清肌酐和尿素氮水平均明显高于假伤组(Z值分别为-2.00、-2.37、-2.62、-2.67、-3.67和-2.34、-3.11、-3.43、-3.11、-3.51,P<0.05)。与电烧伤组伤后0 h比较,电烧伤组大鼠伤后72 h、1周血清肌酐水平均明显升高(P<0.05);与电烧伤组伤后8 h比较,电烧伤组大鼠伤后72 h、1周血清肌酐水平均明显升高(P<0.05);与电烧伤组伤后24 h比较,电烧伤组大鼠伤后1周血清肌酐水平明显升高(P<0.05)。伤后72 h,5组电烧伤大鼠血清肌酐水平相近(P>0.05);与电烧伤组比较,低灯盏花素组、中灯盏花素组、高灯盏花素组大鼠血清尿素氮水平均明显降低(P<0.05);与盐水组比较,中灯盏花素组、高灯盏花素组大鼠血清尿素氮水平均明显降低(P<0.05)。伤后48 h、72 h、1周,电烧伤组大鼠肾组织上清液中CAT活力均明显低于假伤组(Z值分别为-2.22、-2.13、-3.51,P<0.05);伤后8 h、24 h、48 h、72 h、1周,电烧伤组大鼠肾组织上清液中AOPP水平均明显高于假伤组(Z值分别为-2.00、-3.15、-2.71、-2.04、-2.33,P<0.05);伤后0 h~1周,假伤组和电烧伤组大鼠肾组织上清液中Klotho蛋白水平均相近(P>0.05)。与电烧伤组伤后0 h比较,电烧伤组大鼠伤后72 h和1周肾组织上清液中CAT活力及伤后48 h、72 h、1周肾组织上清液中Klotho蛋白水平均明显降低(P<0.05);与电烧伤组伤后8 h比较,电烧伤组大鼠伤后72 h和1周肾组织上清液中CAT活力及伤后48 h、72 h、1周肾组织上清液中Klotho蛋白水平均明显降低(P<0.05);与电烧伤组伤后24 h比较,电烧伤组大鼠伤后72 h和1周肾组织上清液中CAT活力均明显降低(P<0.05);与电烧伤组伤后48 h比较,电烧伤组大鼠伤后1周肾组织上清液中CAT活力明显降低(P<0.05)。伤后72 h,5组电烧伤大鼠肾组织上清液中Klotho蛋白水平相近(P>0.05);与电烧伤组[14.6(12.6,23.6)U/mgprot]比较,低灯盏花素组、中灯盏花素组、高灯盏花素组大鼠肾组织上清液中CAT活力[20.5(18.0,39.8)、24.9(14.7,28.9)、28.0(21.9,39.1)U/mgprot]均明显升高(P<0.05);与盐水组[15.7(13.7,25.6)U/mgprot]比较,中灯盏花素组、高灯盏花素组大鼠肾组织上清液中CAT活力均明显升高(P<0.05);与低灯盏花素组比较,高灯盏花素组大鼠肾组织上清液中CAT活力明显升高(P<0.05);与电烧伤组及盐水组比较,中灯盏花素组、高灯盏花素组大鼠肾组织上清液中AOPP水平均明显降低(P<0.05)。结论高压电烧伤后大鼠肾脏发生氧化应激损伤,且随时间的延长呈进行性加重,应用灯盏花素可减轻高压电烧伤大鼠肾脏氧化应激损伤。
Objective To explore the characteristics of renal oxidative stress injuries in rats with high-voltage electric burns and the intervention effects of breviscapine.Methods This study was an experimental study.One hundred and sixty 8-10-week-old male Sprague Dawley rats were divided into sham injury group,electric burn group,saline group,low breviscapine group,middle breviscapine group,and high breviscapine group,with 60 rats in each of the sham injury group and electric burn group,10 rats in each of the other 4 groups,respectively.The rats in sham injury group and electric burn group were divided into 10 rats at each time point,including post injury hour(PIH)0(immediately),8,24,48,and 72,and post injury week(PIW)1.The rats in sham injury group were not conducted with electrical current to cause sham injury.The rats in the other 5 groups were caused high-voltage electric burns.The rats in sham injury group and electric burn group were not treated after injury.The rats in saline group,low breviscapine group,middle breviscapine group,and high breviscapine group were intraperitoneally injected with 5 mL/kg normal saline or 0.4,1.6,and 4.0 g/L breviscapine,repeated every 24 h until PIH 72.After the model was successfully made,14 rats died,including 1,2,2,and 1 rat(s)at PIH 24,48,and 72 and PIW 1 in electric burn group,4,1,2,and 1 rat(s)at PIH 72 in saline group,low breviscapine group,middle breviscapine group,and high breviscapine group,respectively.The kidney tissue collected from rats in the 6 groups was weighed and the kidney/body weight ratio was calculated.The left upper pole tissue of kidney was collected from each 4 rats in sham injury group,and in electric burn group at PIH 8,24,48,and 72 and PIW 1,and in saline group,low breviscapine group,middle breviscapine group,and high breviscapine group at PIH 72.The renal tubular and renal interstitial injury was evaluated by a semi-quantitative histological scoring system after hematoxylin-eosin staining.The inferior vena cava blood samples were collected from rats in the 6 groups to measure the serum creatinine levels via sarcosine oxidase method,and serum urea nitrogen levels via urease method.The right renal cortices were collected from rats in the 6 groups to measure the catalase(CAT)activity in the supernatant of renal tissue via molybdic acid method,and the levels of advanced oxidation protein product(AOPP)and Klotho protein in the supernatant of renal tissue via enzyme-linked immunosorbent assay.Results At PIH 8,48,and 72 and PIW 1,the kidney/body weight ratios of rats in electric burn group were significantly higher than those in sham injury group(with t values of-0.52,-3.75,-4.05,and-2.25,respectively,P<0.05).At PIH 72,compared with those in electric burn group,saline group,low breviscapine group,and middle breviscapine group,the kidney/body weight ratio of rats in high breviscapine group was significantly decreased(with P values all<0.05).Compared with those in sham injury group,the renal tubular and renal interstitial injury scores of rats in electric burn group at PIH 48 and 72 and PIW 1 were significantly increased(P<0.05).Compared with those in electric burn group at PIH 8 and 24,the renal tubular and renal interstitial injury score of rats in electric burn group at PIW 1 was significantly increased(with P values all<0.05).At PIH 72,the renal tubular and renal interstitial injury scores of rats in the 5 groups of rats with electric burns were similar(P>0.05).At PIH 8,24,48,and 72 and PIW 1,the levels of serum creatinine and serum urea nitrogen of rats in electric burn group were significantly higher than those in sham injury group(with Z values of-2.00,-2.37,-2.62,-2.67,-3.67,-2.34,-3.11,-3.43,-3.11,and-3.51,respectively,P<0.05).Compared with that in electric burn group at PIH 0,the levels of serum creatinine of rats in electric burn group at PIH 72 and PIW 1 were significantly increased(P<0.05).Compared with that in electric burn group at PIH 8,the levels of serum creatinine of rats in electric burn group at PIH 72 and PIW 1 were significantly increased(P<0.05).Compared with that in electric burn group at PIH 24,the level of serum creatinine of rats in electric burn group at PIW 1 was significantly increased(P<0.05).At PIH 72,the levels of serum creatinine of rats in the 5 groups of rats with electric burns were similar(P>0.05).Compared with that in electric burn group,the levels of serum urea nitrogen of rats in low breviscapine group,middle breviscapine group,and high breviscapine group were significantly decreased(P<0.05).Compared with that in saline group,the levels of serum urea nitrogen in middle breviscapine group and high breviscapine group were significantly decreased(P<0.05).At PIH 48 and 72 and PIW 1,the CAT activities in the supernatant of renal tissue of rats in electric burn group were significantly lower than those in sham injury group(with Z values of-2.22,-2.13,and-3.51,respectively,P<0.05).At PIH 8,24,48,and 72 and PIW 1,the levels of AOPP in the supernatant of renal tissue of rats in electric burn group were significantly higher than those in sham injury group(with Z values of-2.00,-3.15,-2.71,-2.04,and-2.33,respectively,P<0.05).At PIH 0-PIW 1,the levels of Klotho protein in the supernatant of renal tissue of rats in sham injury group and electric burn group were all similar(P>0.05).Compared with that in electric burn group at PIH 0,the CAT activities in the supernatant of renal tissue of rats in electric burn group at PIH 72 and PIW 1 and the levels of Klotho protein in the supernatant of renal tissue of rats in electric burn group at PIH 48 and 72 and PIW 1 were significantly decreased(P<0.05).Compared with that in electric burn group at PIH 8,the CAT activities in the supernatant of renal tissue of rats in electric burn group at PIH 72 and PIW 1 and the levels of Klotho protein in the supernatant of renal tissue of rats in electric burn group at PIH 48 and 72 and PIW 1 were significantly decreased(P<0.05).Compared with that in electric burn group at PIH 24,the CAT activities in the supernatant of renal tissue of rats in electric burn group at PIH 72 and PIW 1 were significantly decreased(P<0.05).Compared with that in electric burn group at PIH 48,the CAT activity in the supernatant of renal tissue of rats in electric burn group at PIW 1 was significantly decreased(P<0.05).At PIH 72,the levels of Klotho protein in the supernatant of renal tissue of rats in the 5 groups of rats with electric burns were similar(P<0.05).Compared with 14.6(12.6,23.6)U/mgprot in electric burn group,the CAT activities in the supernatant of renal tissue of rats in low breviscapine group(20.5(18.0,39.8)U/mgprot),middle breviscapine group(24.9(14.7,28.9)U/mgprot),and high breviscapine group(28.0(21.9,39.1)U/mgprot)were significantly increased(P<0.05).Compared with 15.7(13.7,25.6)U/mgprot in saline group,the CAT activities in the supernatant of renal tissue of rats in middle breviscapine group and high breviscapine group were significantly increased(P<0.05).Compared with that in low breviscapine group,the CAT activity in the supernatant of renal tissue of rats in high breviscapine group was significantly increased(P<0.05).Compared with those in electric burn group and saline group,the levels of AOPP in the supernatant of renal tissue of rats in middle breviscapine group and high breviscapine group were significantly decreased(P<0.05).Conclusions After high-voltage electric burns,oxidative stress injury occur in the kidneys of rats,which is aggravated with time extension.Breviscapine can alleviate oxidative stress injuries in the kidneys of rats with high-voltage electric burns.
作者
李聪颖
赵学刚
郝嘉文
盖晨阳
鲁梦远
张晶
张庆富
冯建科
屠丽红
Li Congying;Zhao Xuegang;Hao Jiawen;Ge Chenyang;Lu Mengyuan;Zhang Jing;Zhang Qingfu;FengJianke;TuLihong(Burn and Wound Repair Center,the Third Hospital of Hebei Medical University,Hebei Burn Treatment Technology Innovation Center,Shijiazhuang 050051,China)
出处
《中华烧伤与创面修复杂志》
CAS
CSCD
北大核心
2024年第8期746-755,共10页
Chinese Journal of Burns And Wounds
基金
河北省自然科学基金面上项目(H2024206495)
河北省创新能力提升计划项目(225676120H)。
关键词
烧伤
电
氧化性应激
肾
晚期氧化蛋白产物
过氧化氢酶
灯盏花素
Burns,electric
Oxidative stress
Kidney
Advanced oxidation protein products
Catalase
Breviscapine
