大黄素对小鼠肾脏毒性表现的凋亡机制
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摘要
目的:观察大黄素长期给药对小鼠肾脏毒性的影响,探讨其可能的毒性机制,为临床合理用药及进一步研究提供一定依据。方法:昆明种小鼠30只,随机分为空白组和大黄素低、高剂量(0.8,1.6 g·kg~(-1))组,每组10只,雌雄各半。连续灌胃给药11周。给药期间,观察记录小鼠的一般情况;停止给药后检测动物血清尿素氮(BUN),肌酐(SCr),丙二醛(MDA),超氧化物歧化酶(SOD),肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)等指标。计算肾脏指数并检测肾脏中谷胱甘肽过氧化物酶(GSH-Px)和还原型谷胱甘肽/氧化型谷胱甘肽(GSH/GSSG),取肾脏进行病理组织学检查,免疫组化法检测肾脏转化生长因子-β_1(TGF-β_1)及半胱氨酸蛋白酶-3(Caspase-3)蛋白表达。结果:与同性别空白组比较,给药组小鼠体质量下降明显,肾脏指数降低,BUN和SCr升高明显(P<0.05,P<0.01),SOD活力明显降低(P<0.05,P<0.01),TNF-α含量明显增多(P<0.05,P<0.01),大黄素高剂量组GSH-Px含量和GSH/GSSG明显下降(P<0.05),Caspase-3表达明显上升(P<0.05)。肾脏病理组织学检测,大黄素高剂量组可见肾小管上皮细胞肿胀、肾小管管腔中蛋白管型、充血和淋巴细胞小灶性增生明显,大黄素低剂量组以上表现较高剂量组不明显。结论:大黄素长期大剂量给药对小鼠肾脏存在一定毒性,在1.6 g·kg~(-1)·d~(-1)剂量下毒性作用明显,无明显性别差异。其潜在毒性机制可能为引起氧化系统紊乱,诱发氧化应激损伤,触发炎症反应,进而使细胞凋亡。
Objective:To observe the effect of long-term administration of emodin on the kidney toxicity of mice,explore its possible toxic mechanism,and provide some basis for clinical rational drug use and further research.Method:The 30 Kunming mice,half male and half female,were randomly divided into 3 groups:control group,emodin low dose group and emodin high dose groups(0.8,1.6 g·kg~(-1)),10 mice in each group.Continuous intragastric administration was given for 11 weeks.During administration,the general situation of the mice was observed and recorded.After treatment,the serum urea nitrogen(BUN),creatinine(SCr),malondialdehyde(MDA),superoxide dismutase(SOD)tumor necrosis factor(TNF-α)and interleukin-6(IL-6)were detected.Kidney index was calculated and glutathione peroxidase(GSH-Px)and reduced glutathione/oxidized glutathione(GSH/GSSG)ratio were measured.The kidneys were taken for histopathological examination and the protein expression levels of transforming growth factor-β_1(TGF-β_1)and cysteine aspartic acid specific protease-3(Caspase-3)were then detected by immunohistochemistry assay.Result:As compared with control group of the same sex,the weight of mice in the administration groups was decreased significantly,renal index was decreased while BUN and SCr levels were increased significantly(P<0.05,P<0.01);activity of SOD was decreased significantly(P<0.05,P<0.01);content of TNF-αwas increased significantly(P<0.05,P<0.01),while the content of GSH-Px and the ratio of GSH/GSSG were decreased significantly in high emodin group(P<0.05),and the expression of Caspase-3 was increased significantly in high emodin group(P<0.05).Renal histopathology:tubule epithelial cells swelling,tubulointerstitial protein tube type,hyperemia and lymphocytic small focal hyperplasia were observed in emodin high dose group,but the performance above was not obvious in low dose group.Conclusion:The long-term administration of emodin at a large dose would show toxicity effect on mice kidney,and the toxicity was obvious at the dose of 1.6 g·kg~(-1)·d~(-1),but there was no significant difference between the sexes.The mechanism of its potential toxicity may be related to the disorder of oxidation system,the injury of oxidative stress,the triggering of inflammatory reaction,and the apoptosis of cells.
Objective:To observe the effect of long-term administration of emodin on the kidney toxicity of mice,explore its possible toxic mechanism,and provide some basis for clinical rational drug use and further research.Method:The 30 Kunming mice,half male and half female,were randomly divided into 3 groups:control group,emodin low dose group and emodin high dose groups(0.8,1.6 g·kg~(-1)),10 mice in each group.Continuous intragastric administration was given for 11 weeks.During administration,the general situation of the mice was observed and recorded.After treatment,the serum urea nitrogen(BUN),creatinine(SCr),malondialdehyde(MDA),superoxide dismutase(SOD)tumor necrosis factor(TNF-α)and interleukin-6(IL-6)were detected.Kidney index was calculated and glutathione peroxidase(GSH-Px)and reduced glutathione/oxidized glutathione(GSH/GSSG)ratio were measured.The kidneys were taken for histopathological examination and the protein expression levels of transforming growth factor-β_1(TGF-β_1)and cysteine aspartic acid specific protease-3(Caspase-3)were then detected by immunohistochemistry assay.Result:As compared with control group of the same sex,the weight of mice in the administration groups was decreased significantly,renal index was decreased while BUN and SCr levels were increased significantly(P<0.05,P<0.01);activity of SOD was decreased significantly(P<0.05,P<0.01);content of TNF-αwas increased significantly(P<0.05,P<0.01),while the content of GSH-Px and the ratio of GSH/GSSG were decreased significantly in high emodin group(P<0.05),and the expression of Caspase-3 was increased significantly in high emodin group(P<0.05).Renal histopathology:tubule epithelial cells swelling,tubulointerstitial protein tube type,hyperemia and lymphocytic small focal hyperplasia were observed in emodin high dose group,but the performance above was not obvious in low dose group.Conclusion:The long-term administration of emodin at a large dose would show toxicity effect on mice kidney,and the toxicity was obvious at the dose of 1.6 g·kg~(-1)·d~(-1),but there was no significant difference between the sexes.The mechanism of its potential toxicity may be related to the disorder of oxidation system,the injury of oxidative stress,the triggering of inflammatory reaction,and the apoptosis of cells.
引文
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[18]兰美华,吴红彦.中药植物多糖抗氧化作用研究进展[J].实用中医药杂志,2012,28(4):326-327.
[19]汪志峰,仲崇俊,张毅,等.缺血后处理对大鼠局灶性脑缺血再灌注损伤保护作用的实验研究[J].南通大学学报:医学版,2012,32(6):504-506,440.
[20]XIONG J,SUN Q,JI K,et al. Epidermal growth factor promotes transforming growth factor-β1-induced epithelial-mesenchymal transition in HK-2 cells through a synergistic effect on Snail[J]. Mol Biol Rep,2013,41(1):241-250.
[21]MENG X M,HUANG X R,XIAO J,et al. Diverse roles of TGF-βreceptorⅡin renal fibrosis and inflammation in vivo and in vitro[J]. J Pathol,2012,227(2):175-188.
[22]Dizin E,Hasler U,Nlandukhodo S,et al. Albuminuria induces a proinflammatory and profibrotic response in cortical collecting ducts via the 24p3 receptor[J]. Am J Physiol Renal,2013,305(7):1053-1063.
[23]何文俊,张涛,蒋学俊,等.炎症因子与慢性心力衰竭[J].中华临床医师杂志:电子版,2011,5(2):457-460.
[2]MA J W,HUANG C M,LIN Y C,et al. Aloe-emodin inhibits HER-2 expression through the downregulation of Y-box binding protein-1 in HER-2-overexpressing human breast cancer cells[J]. Oncotarget,2016,7(37):58915-58930.
[3]张耀雷,李昆,杨炯,等.大黄素缓解球囊损伤致大鼠颈动脉狭窄及其机制[J].第三军医大学学报,2017,39(1):48-53.
[4]XUE J,CHEN F,WANG J,et al. Emodin protects against concanavalin a-induced hepatitis in mice through inhibiting activation of the p38 MAPK-NF-κB signaling pathway[J]. Cell Physiol Biochem,2015,35(4):1557-1570.
[5]王少杰,李晓洁,徐志猛,等.大黄素对小鼠急性脂肪肝的改善作用[J].中国药科大学学报,2017,48(1):89-95.
[6]孙禄,袁迅玲,杨亚琦,等.大黄素对大鼠肾纤维化肾间质中骨形态发生蛋白-7表达的影响及意义[J].中医临床研究,2017,9(3):73-74,79.
[7]赵玲,胡昌江,潘新,等.长期服用生大黄、熟大黄对大鼠肝肾功能影响的比较[J].中国医院药学杂志,2015,35(15):1384-1387.
[8]邓诺,易艳,梁爱华,等.大黄肾脏毒性部分机制研究[J].中国中药杂志,2018,43(13):2777-2783.
[9]王青秀.大黄及其主要成份的毒性毒理研究[D].北京:中国人民解放军军事医学科学院,2007.
[10]WANG C,DAI X, LIU H, et al. Involvement of PPARγin emodin-induced HK-2 cell apoptosis[J].Toxicol in Vitro,2015,29(1):228-233.
[11]雷瑶,王志洁,邱国福,等.大黄素脂质体对Hep-2细胞毒的检测[J].武汉大学学报:理学版,2006,52(2):247-251.
[12]王翔飞,葛均波,孙爱军,等.大黄素通过p53途径抑制血管平滑肌细胞增殖的实验研究[J].中华心血管病杂志,2006,34(1):44-49.
[13]陈文,张祎,王沛明,等.大黄对内毒素血症模型大鼠氧化应激损伤的保护作用初探[J].中国实验方剂学杂志,2016,22(15):112-116.
[14]刘德明,周春燕,吴嘉思,等.大黄素通过线粒体通路诱导Hep G2细胞凋亡[J].中国实验方剂学杂志,2018,24(3):104-108.
[15]刘德明,周春燕,吴嘉思,等.大黄素通过激活线粒体caspase-8通路诱导L02细胞凋亡[J].中药药理与临床,2017,33(5):23-26.
[16]谭汉全,尹雪梅. PTH、BUN及SCr联合检测在肾功能损伤早期诊断中的临床价值[J].海南医学院学报,2015,21(2):197-199.
[17]YANG Y,LIU P,CHEN L,et al. Therapeutic effect of Ginkgo biloba polysaccharide in rats with focal cerebral ischemia/reperfusion(I/R)injury[J]. Carbohydr Polym,2013,98(2):1383-1388.
[18]兰美华,吴红彦.中药植物多糖抗氧化作用研究进展[J].实用中医药杂志,2012,28(4):326-327.
[19]汪志峰,仲崇俊,张毅,等.缺血后处理对大鼠局灶性脑缺血再灌注损伤保护作用的实验研究[J].南通大学学报:医学版,2012,32(6):504-506,440.
[20]XIONG J,SUN Q,JI K,et al. Epidermal growth factor promotes transforming growth factor-β1-induced epithelial-mesenchymal transition in HK-2 cells through a synergistic effect on Snail[J]. Mol Biol Rep,2013,41(1):241-250.
[21]MENG X M,HUANG X R,XIAO J,et al. Diverse roles of TGF-βreceptorⅡin renal fibrosis and inflammation in vivo and in vitro[J]. J Pathol,2012,227(2):175-188.
[22]Dizin E,Hasler U,Nlandukhodo S,et al. Albuminuria induces a proinflammatory and profibrotic response in cortical collecting ducts via the 24p3 receptor[J]. Am J Physiol Renal,2013,305(7):1053-1063.
[23]何文俊,张涛,蒋学俊,等.炎症因子与慢性心力衰竭[J].中华临床医师杂志:电子版,2011,5(2):457-460.