五味子乙素对缺血再灌注致小鼠急性肾损伤的影响
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摘要
目的观察五味子乙素(SchB)对肾缺血再灌注致小鼠急性肾损伤的影响,并探讨其可能作用机制。方法将45只C57BL/6小鼠随机分成假手术组、模型组和五味子乙素组,每组15只。模型组和五味子乙素组小鼠采用夹闭双侧肾蒂40 min以建立肾缺血再灌注模型,五味子乙素组造模前用80 mg/(kg·d)的SchB预先连续灌胃给药7天。各组于造模后第1、2、7天检测小鼠血肌酐(Scr)与尿素氮(BUN),PCR法检测肾组织中肾损伤分子(Kim-1)、嗜中性粒细胞明胶酶相关载脂蛋白(NGAL) mRNA表达,Western blot法检测肾组织中天冬氨酸特异性半胱氨酸蛋白酶(Caspase-3)、B细胞淋巴瘤/白血病基因2 (Bcl-2)、Bcl-2相关X蛋白(Bax)蛋白表达,HE染色观察肾组织的病理变化,原位末端标记法(TUNEL)检测肾小管细胞的凋亡情况。结果第1、2天,与假手术组比较,模型组肾组织凋亡指数、Caspase-3蛋白表达、血Scr与BUN水平及肾组织中Kim-1、NGAL mRNA表达均显著升高(P <0. 05或P <0. 01),肾组织的病理损伤明显加重,Bcl-2/Bax值降低(P <0. 01)。与模型组比较,五味子乙素组凋亡指数、Bcl-2/Bax值、Caspase-3蛋白表达,血Scr与BUN水平及肾组织中Kim-1、NGAL mRNA表达均降低(P <0. 05或P <0. 01),肾组织的病理损伤明显减轻。第7天,模型组与五味子乙素组各项观察指标差异均无统计学意义(P> 0. 05)。结论 SchB对肾缺血再灌注所致急性肾损伤小鼠的肾功能、病理损伤及细胞凋亡情况有一定的改善作用,其作用机制可能与下调肾组织Caspase-3和上调Bcl-2/Bax值有关。
Objective To observe the effects of schisandrin B( SchB) on acute kidney injury induced by renal ischemia-reperfusion( I/R) in mice,and to explore its possible mechanism. Methods A total of 45 C57 BL/6 mice were randomly divided into sham operation group,model group and SchB group,with 15 mice in each group.The model group and SchB group were treated with clamping bilateral renal pedicles for 40 min to establish I/R model. Before modeling,SchB group was pretreated with 80 mg/( kg·d) of SchB by gavage for 7 days. The levels of serum creatinine( Scr) and urea nitrogen( BUN) of each group were detected on the 1 st,2 nd and 7 th day after modeling. The expression of kidney injury molecule 1( Kim-1) and neutrophil gelatinase-associated lipocalin( NGAL)mRNA in kidney tissues were detected by PCR. The expression of cysteine-containing aspartate-specific proteases 3( caspase-3),B cell lymphoma/promyelocytic leukemia( Bcl-2) and Bcl-2 related protein were detected by Western blot. HE staining was used to observe the pathological changes of kidney tissues. Terminal-deoxynucleoitidyl Transferase Mediated Nick End Labeling( TUNEL) was used to detect the apoptosis of renal tubular cell. Results On the1 st and 2 nd day,compared with sham operation group,renal tissue apoptosis index,Caspase-3 protein expression,blood Scr and BUN levels,and expression of Kim-1 and NGAL mRNA in renal tissue were significantly increased in the model group( P < 0. 05 or P < 0. 01). The pathological damage of renal tissue was obviously aggravated. The Bcl-2/Bax value decreased( P < 0. 01). Compared with the model group,the apoptotic index,Bcl-2/Bax value,Caspase-3 protein expression,blood Scr and BUN levels and the expression of Kim-1 and NGAL mRNA in renal tissue were decreased in SchB group( P < 0. 05 or P < 0. 01),and the pathological damage of renal tissue was significantly alleviated. On the 7 th day,there was no significant difference between the model group and the SchB group( P >0. 05). Conclusion SchB can improve the renal function,pathological damage and apoptosis of acute kidney injury induced by ischemia-reperfusion. It may be related to down-regulation of caspase-3 and up-regulation of Bcl-2/Bax ratio.
Objective To observe the effects of schisandrin B( SchB) on acute kidney injury induced by renal ischemia-reperfusion( I/R) in mice,and to explore its possible mechanism. Methods A total of 45 C57 BL/6 mice were randomly divided into sham operation group,model group and SchB group,with 15 mice in each group.The model group and SchB group were treated with clamping bilateral renal pedicles for 40 min to establish I/R model. Before modeling,SchB group was pretreated with 80 mg/( kg·d) of SchB by gavage for 7 days. The levels of serum creatinine( Scr) and urea nitrogen( BUN) of each group were detected on the 1 st,2 nd and 7 th day after modeling. The expression of kidney injury molecule 1( Kim-1) and neutrophil gelatinase-associated lipocalin( NGAL)mRNA in kidney tissues were detected by PCR. The expression of cysteine-containing aspartate-specific proteases 3( caspase-3),B cell lymphoma/promyelocytic leukemia( Bcl-2) and Bcl-2 related protein were detected by Western blot. HE staining was used to observe the pathological changes of kidney tissues. Terminal-deoxynucleoitidyl Transferase Mediated Nick End Labeling( TUNEL) was used to detect the apoptosis of renal tubular cell. Results On the1 st and 2 nd day,compared with sham operation group,renal tissue apoptosis index,Caspase-3 protein expression,blood Scr and BUN levels,and expression of Kim-1 and NGAL mRNA in renal tissue were significantly increased in the model group( P < 0. 05 or P < 0. 01). The pathological damage of renal tissue was obviously aggravated. The Bcl-2/Bax value decreased( P < 0. 01). Compared with the model group,the apoptotic index,Bcl-2/Bax value,Caspase-3 protein expression,blood Scr and BUN levels and the expression of Kim-1 and NGAL mRNA in renal tissue were decreased in SchB group( P < 0. 05 or P < 0. 01),and the pathological damage of renal tissue was significantly alleviated. On the 7 th day,there was no significant difference between the model group and the SchB group( P >0. 05). Conclusion SchB can improve the renal function,pathological damage and apoptosis of acute kidney injury induced by ischemia-reperfusion. It may be related to down-regulation of caspase-3 and up-regulation of Bcl-2/Bax ratio.
引文
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[12]CAI Z,LIU J,BIAN H,et al. Suppression of P2X7/NF-κB pathways by Schisandrin B contributes to attenuation of lipopolysaccharide-induced inflammatory responses in acute lung injury[J]. Arch Pharm Res,2016,39(4):499-507.
[13]吴银娜.五味子乙素干预顺铂诱导的肾小管上皮细胞凋亡与自噬的研究[D].天津:天津医科大学,2015.
[14]HAVASI A,BORKAN SC. Apoptosis and acute kidney injury[J]. Kidney Int,2011,80(1):29-40.
[15]周奥飞,倪钊,李强,等. 17β-雌二醇对大鼠肾缺血再灌注损伤肾小管细胞凋亡及炎症的影响[J].临床与实验病理学杂志,2018,34(3):300-305.
[16]VISCONTI R,D'ADAMIO L. Functional cloning of genes regulating apoptosis in neuronal cells[J]. Methods Mol Biol,2007,399:125-131. doi:10. 1007/978-1-59745-504-6_9.
[17]RENHUA L,MIAOLIN C,JUNLIN W,et al. The level of the biomarkers at the time of nephrology consultation might predict the prognosis of acute kidney injury in hospitalized patients[J]. Blood Purif,2014,38(2):89-95.
[18]ICHIMURA T,BONVENTRE JV,BAILLY V,et al. Kidney injury molecule-1(KIm-1),a putative epithelial cell adhesion molecule containing a novel immunoglobulin domain,is up-regulated in renal cells after injury[J]. J Biol Chem,1998,273(7):4135-4142.
[2]SCHORTGEN F,ASFAR P. Update in sepsis and acute kidney injury 2014[J]. Am J Respir Crit Care Med,2015,191(11):1226-1231.
[3]SUSANTITAPHONG P,CRUZ D N,CERDA J,et al.World incidence of AKI:a meta-analysis[J]. Clin J Am Soc Nephrol,2013,8(9):1482-1493.
[4]WAIKAR SS,CURHAN GC,WALD R,et al. Declining mortality in patients with acute renal failure,1988 to 2002[J]. J Am Soc Nephrol,2006,17(4):1143-1150.
[5]COCA SG,SINGANAMALA S,PARIKH CR. Chronic kidney disease after acute kidney injury:a systematic review and meta-analysis[J]. Kidney Int,2012,81(5):442-448.
[6]KAUSHAL GP,SHAH SV. Challenges and advances in the treatment of AKI[J]. J Am Soc Nephrol,2014,25(5):877-883.
[7]闫荟羽,周微,曲晓宇,等.五味子乙素的药理研究进展[J].中国生化药物杂志,2014,34(2):153-156.
[8]LAI Q,LUO Z,WU C,et al. Attenuation of cyclosporine A induced nephrotoxicity by schisandrin B through suppression of oxidative stress,apoptosis and autophagy[J]. Int Immunopharmacol,2017,52:15-23. doi:10. 1016/j. intimp. 2017. 08. 019.
[9]BIENHOLZ A,REIS J,SANLI P,et al. Citrate shows protective effects on cardiovascular and renal function in ischemia-induced acute kidney injury[J]. BMC Nephrol,2017,18(1):130.
[10]ANDREOLI SP,MCATEER JA,MALLETT C. Reactive oxygen molecule-mediated injury in endothelial and renal tubular epithelial cells in vitro[J]. Kidney Int,1990,38(5):785-794.
[11]HAVASI A,BORKAN SC. Apoptosis and acute kidney injury[J]. Kidney Int,2011,80(1):29-40.
[12]CAI Z,LIU J,BIAN H,et al. Suppression of P2X7/NF-κB pathways by Schisandrin B contributes to attenuation of lipopolysaccharide-induced inflammatory responses in acute lung injury[J]. Arch Pharm Res,2016,39(4):499-507.
[13]吴银娜.五味子乙素干预顺铂诱导的肾小管上皮细胞凋亡与自噬的研究[D].天津:天津医科大学,2015.
[14]HAVASI A,BORKAN SC. Apoptosis and acute kidney injury[J]. Kidney Int,2011,80(1):29-40.
[15]周奥飞,倪钊,李强,等. 17β-雌二醇对大鼠肾缺血再灌注损伤肾小管细胞凋亡及炎症的影响[J].临床与实验病理学杂志,2018,34(3):300-305.
[16]VISCONTI R,D'ADAMIO L. Functional cloning of genes regulating apoptosis in neuronal cells[J]. Methods Mol Biol,2007,399:125-131. doi:10. 1007/978-1-59745-504-6_9.
[17]RENHUA L,MIAOLIN C,JUNLIN W,et al. The level of the biomarkers at the time of nephrology consultation might predict the prognosis of acute kidney injury in hospitalized patients[J]. Blood Purif,2014,38(2):89-95.
[18]ICHIMURA T,BONVENTRE JV,BAILLY V,et al. Kidney injury molecule-1(KIm-1),a putative epithelial cell adhesion molecule containing a novel immunoglobulin domain,is up-regulated in renal cells after injury[J]. J Biol Chem,1998,273(7):4135-4142.