健脾清化方对急性肾缺血再灌注SD大鼠氧化应激损伤的影响
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摘要
目的:探讨健脾清化方对急性肾缺血再灌注SD大鼠氧化应激损伤的影响。方法:将40只SD大鼠随机分成正常组、急性肾缺血再灌注模型组、健脾清化方干预组和盐酸罗格列酮干预组,每组10只。健脾清化方干预组给予健脾清化方灌胃(19. 3 g/kg,20倍人体剂量),盐酸罗格列酮组给予罗格列酮灌胃(1. 3 mg/kg,20倍人体剂量),其他组给予生理盐水,1次/d,连续给药1周后,造模并取材。观察各组SD大鼠血肌酐(Scr)、尿素氮(BUN)水平,观察各组大鼠血清超氧化物歧化酶(SOD)、一氧化氮(NO)、一氧化氮合酶(NOS)、羟自由基(OH)抑制能力的水平,ELISA检测各组大鼠血清IL-1β、IL-10水平,免疫印迹法观察各组大鼠肾组织TNF-α、NF-κB蛋白表达水平。结果:与正常组比较,模型组的Scr、BUN均显著增加(P <0. 05),抑制羟自由基能力、NOS、NO水平提升(P <0. 05),SOD水平明显下降(P <0. 05)。与模型组比较,健脾清化方干预组及罗格列酮干预组Scr、BUN(P <0. 05)水平均明显降低,NOS(P <0. 05)、NO显著下降,抑制羟自由基能力与SOD水平显著提升(P <0. 05)。模型组、盐酸罗格列酮组与健脾清化方组IL-1β、IL-10水平均较正常组明显升高(P <0. 05),与模型组比较,盐酸罗格列酮组与健脾清化方组的IL-1β水平更低,IL-10的水平更高。盐酸罗格列酮组的IL-1β水平比健脾清化方组更高,IL-10水平更低。与正常组比较,模型组、盐酸罗格列酮组与健脾清化方组大鼠肾组织的NF-κB、TNF-α蛋白表达量均显著提升。与模型组比较,盐酸罗格列酮组与健脾清化方组NF-κB、TNF-α蛋白表达量均显著下调。与盐酸罗格列酮组比较,健脾清化方组的NF-κB、TNF-α蛋白表达量更低。结论:健脾清化方可明通过降低NO、NOS水平,增强SOD和抑制OH自由基能力,从而延缓急性肾缺血再灌注SD大鼠氧化应激损伤水平,抑制炎性反应,从而减少对肾脏的损伤。
Objective: To investigate the effects of Jianpi Qinghua decoction on oxidative stress injury in SD rats with acute renal ischemia reperfusion. Methods: A total of 40 SD rats were randomly divided into a normal group( n = 10),an acute renal ischemiareperfusion model group( n = 10),a Jianpi Qinghua decoction intervention group( n = 10),a rosiglitazone hydrochloride intervention group( n = 10). The Jianpi Qinghua decoction intervention group was given Jianpi Qinghua recipe( 19. 3 mg·kg-1,20 times human dose),and the rosiglitazone hydrochloride intervention group was given rosiglitazone( 1. 3 mg·kg-1,20 times human dose). The other groups were given normal saline once a day,and after one week of continuous administration. The rats was molded and the materials were taken. The levels of serum creatinine( Scr) and urea nitrogen( BUN) in SD rats were observed. The serum superoxide dismutase( SOD),nitric oxide( NO),nitric oxide synthase( NOS) were observed in each group. The levels of hydroxyl-free radical( OH) inhibition ability,serum IL-1β,IL-10 levels in each group were detected by ELISA. The expressions of TNF-α and NF-KB in renal tissues of each group were observed by Western blot. Results: Compared with the normal group,the Scr and BUN of the model group were significantly increased( P < 0. 05). The ability of inhibiting hydroxyl radicals,the levels of NOS and NO were significantly increased( P < 0. 05),and the level of SOD was significantly decreased( P < 0. 05). Compared with the model group,the Scr and BUN levels in the Jianpi Qinghua decoction group and the rosiglitazone treatment group were significantly decreased( P <0. 05),NOS( P < 0. 05) and NO decreased significantly,and the level of ability to inhibit hydroxyl free radicals and SOD was significantly increased( P < 0. 05). The levels of IL-1β and IL-10 in the model group,the rosiglitazone hydrochloride group and the Jianpi Qinghua decoction group were significantly increased than those in the normal group( P < 0. 05). Compared with the model group,the level of IL-1β in the rosiglitazone hydrochloride group and the Jianpi Qinghua decoction group was lower and the level of IL-10 was higher. The level of IL-1β in the rosiglitazone hydrochloride group was higher than that in the Jianpi Qinghua decoction group while the IL-10 level was lower. Compared with the normal group,the expression of NF-KB and TNF-α protein in the renal tissue of the model group,rosiglitazone hydrochloride group and Jianpi Qinghua decoction group were significantly increased. Compared with the model group,the expression levels of NF-KB and TNF-α protein in the rosiglitazone hydrochloride group and Jianpi Qinghua decoction group were significantly down-regulated. Compared with the rosiglitazone hydrochloride group,the expression of NF-KB and TNF-α protein in Jianpi Qinghua decoction group was lower. Conclusion: Jianpi Qinghua decoction can reduce the levels of NO and NOS,enhance SOD and inhibit OH free radicals,thereby delaying the level of oxidative stress injury in SD rats with acute renal ischemia-reperfusion and inhibiting inflammatory reaction,thereby reducing the kidney damage.
Objective: To investigate the effects of Jianpi Qinghua decoction on oxidative stress injury in SD rats with acute renal ischemia reperfusion. Methods: A total of 40 SD rats were randomly divided into a normal group( n = 10),an acute renal ischemiareperfusion model group( n = 10),a Jianpi Qinghua decoction intervention group( n = 10),a rosiglitazone hydrochloride intervention group( n = 10). The Jianpi Qinghua decoction intervention group was given Jianpi Qinghua recipe( 19. 3 mg·kg-1,20 times human dose),and the rosiglitazone hydrochloride intervention group was given rosiglitazone( 1. 3 mg·kg-1,20 times human dose). The other groups were given normal saline once a day,and after one week of continuous administration. The rats was molded and the materials were taken. The levels of serum creatinine( Scr) and urea nitrogen( BUN) in SD rats were observed. The serum superoxide dismutase( SOD),nitric oxide( NO),nitric oxide synthase( NOS) were observed in each group. The levels of hydroxyl-free radical( OH) inhibition ability,serum IL-1β,IL-10 levels in each group were detected by ELISA. The expressions of TNF-α and NF-KB in renal tissues of each group were observed by Western blot. Results: Compared with the normal group,the Scr and BUN of the model group were significantly increased( P < 0. 05). The ability of inhibiting hydroxyl radicals,the levels of NOS and NO were significantly increased( P < 0. 05),and the level of SOD was significantly decreased( P < 0. 05). Compared with the model group,the Scr and BUN levels in the Jianpi Qinghua decoction group and the rosiglitazone treatment group were significantly decreased( P <0. 05),NOS( P < 0. 05) and NO decreased significantly,and the level of ability to inhibit hydroxyl free radicals and SOD was significantly increased( P < 0. 05). The levels of IL-1β and IL-10 in the model group,the rosiglitazone hydrochloride group and the Jianpi Qinghua decoction group were significantly increased than those in the normal group( P < 0. 05). Compared with the model group,the level of IL-1β in the rosiglitazone hydrochloride group and the Jianpi Qinghua decoction group was lower and the level of IL-10 was higher. The level of IL-1β in the rosiglitazone hydrochloride group was higher than that in the Jianpi Qinghua decoction group while the IL-10 level was lower. Compared with the normal group,the expression of NF-KB and TNF-α protein in the renal tissue of the model group,rosiglitazone hydrochloride group and Jianpi Qinghua decoction group were significantly increased. Compared with the model group,the expression levels of NF-KB and TNF-α protein in the rosiglitazone hydrochloride group and Jianpi Qinghua decoction group were significantly down-regulated. Compared with the rosiglitazone hydrochloride group,the expression of NF-KB and TNF-α protein in Jianpi Qinghua decoction group was lower. Conclusion: Jianpi Qinghua decoction can reduce the levels of NO and NOS,enhance SOD and inhibit OH free radicals,thereby delaying the level of oxidative stress injury in SD rats with acute renal ischemia-reperfusion and inhibiting inflammatory reaction,thereby reducing the kidney damage.
引文
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[2]任冰霜,雷艳,黄梁浒.肾脏缺血再灌注损伤的分子机制[J].实用医学杂志,2016,32(10):1710-1712.
[3]罗京.PPARγ激动剂罗格列酮对大鼠肾脏再灌注损伤的保护作用[J].海南医学,2017,28(12):1901-1904.
[4]邹赟,朱祎,邵命海,等.健脾清化方对5/6肾切除大鼠ATII/NADPH氧化应激通路的干预作用[J].中南大学学报:医学版,2013,38(8):779-784.
[5]张蕾,梁立峰.高糖环境下肾小球内皮细胞损伤与糖尿病肾病的关系[J].现代临床医学,2018,44(2):148-151.
[6]杨雅楠.大蒜素对慢性肾衰大鼠氧化应激和炎性反应的影响[J].实用药物与临床,2018,21(1):24-28.
[7]Weiss MF,Scivittaro V,Anderson JM.Oxidative stress and increased expression of growth factors in lesions of failed hemodialysis access[J].Am J Kidney Dis,2001,37(5):970-980.
[8]Guo J,Guan Q,Liu X,et al.Relationship of clusterin with renal inflammation and fibrosis after the recovery phase of ischemia-reperfusion injury[J].BMC Nephrol,2016,17(1):133.
[9]麻志恒,彭文,倪兆慧,等.健脾清化方治疗慢性肾脏病(3期)脾虚湿热型患者的临床疗效观察[J].中华中医药杂志,2016,31(10):4333-4337.
[10]王新陆.血浊证的辨证治疗[J].山东中医药,2007,26(1):3-5.
[11]孙大伟,高青,齐新.从脾论治心肌缺血再灌注损伤无复流现象[J].山东中医杂志,2017,36(8):635-637,641.
[12]徐艳秋,杨超茅,顾向晨,等.慢性肾衰竭早期从脾论治的临床疗效分析[J].中国中西医结合肾病杂志,2016,17(6):502-505.
[13]赵维,吴广礼.补气类单味中药及其提取物防治急性肾损伤研究进展[J].解放军医药杂志,2016,28(4):109-113.
[14]谭婷婷.补脾肾活血法对糖尿病肾病患者氧化蛋白产物的研究[D].广州:广州中医药大学,2015.