白藜芦醇联合内皮祖细胞移植修复镉致大鼠的肾损伤
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摘要
背景:研究表明白藜芦醇能有效提高大鼠近曲小管上皮细胞存活率,抑制细胞凋亡,对肾小管上皮细胞损伤具有保护作用,但白藜芦醇治疗大鼠肾损伤的具体机制尚有待进一步明确。目的:探讨白藜芦醇联合内皮祖细胞移植对慢性镉暴露致大鼠肾脏损伤的保护作用及机制。方法:体外复苏培养中国医学科学院提供的大鼠内皮祖细胞,并制备其细胞悬液,对内皮祖细胞行CM-Dil荧光标记。将购自北京维通利华动物实验技术有限公司的Wistar大鼠随机分5组,对照组皮下注射生理盐水,其他4组均皮下注射6μmol/kg的CdCl_2,每日1次,连续染毒6周,建立肾损伤大鼠模型。建模后模型组大鼠尾静脉注射0.5 mL的L-DMEM,内皮祖细胞组大鼠尾静脉注射0.5 mL的1×10~(10) L~(-1)内皮祖细胞组悬液;白藜芦醇组大鼠灌胃120 mg/kg白藜芦醇,联合组大鼠同时给予以上2种干预,每日1次,连续治疗7 d。结果与结论:①血清及尿液指标检测:与模型组比较,白藜芦醇组、内皮祖细胞组及联合组尿素氮、肌酐、尿乙酰氨基葡糖苷酶、尿γ-L-谷氨酰转肽酶及尿蛋白水平均降低(P <0.05),联合组降低更明显(P <0.01);②肾脏组织指标检测:与模型组比较,白藜芦醇组、内皮祖细胞组及联合组肾皮质中活性氧及丙二醛水平降低(P <0.05),超氧化物歧化酶和谷胱甘肽过氧化物酶活力升高(P <0.05),联合组上述指标变化更为明显(P <0.01);③肾脏组织病理变化:联合组的肾脏组织病理改善情况明显优于内皮祖细胞组、白藜芦醇及模型组,荧光显微镜观察显示,联合组CM-Dil标记的内皮祖细胞多于内皮祖细胞组(P <0.05);④结果证实,白藜芦醇联合内皮祖细胞移植对慢性镉暴露致大鼠肾脏损伤具有保护作用。
BACKGROUND: Studies have shown that resveratrol can effectively improve the survival rate of rat proximal tubular epithelial cells and inhibit cell apoptosis to protect against renal tubular epithelial cell injury. However, the specific mechanism of resveratrol in the treatment of renal injury in rats remains to be further clarified.OBJECTIVE: To investigate the protective effect and mechanism of resveratrol combined with endothelial progenitor cell transplantation on renal injury caused by chronic cadmium exposure in rats.METHODS: Rat endothelial progenitor cells provided by the Chinese Academy of Medical Sciences were resuscitated and cultured in vitro,and cell suspensions were then prepared. Endothelial progenitor cells were labeled with CM-Dil. Wistar rats purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. in China were randomly divided into control group, pure cadmium poisoning group(model group),resveratrol group, endothelial progenitor cell group and combined treatment group, with 15 rats in each group. In the control group,physiological saline was injected subcutaneously. In the other four groups, the rats were subcutaneously administered with 6 μmol/kg cadmium chloride once daily for 6 weeks to establish animal models of renal injury. After modeling, the rat models in the latter four groups were given 0.5 mL of L-DMEM via the tail vein, 0.5 mL of endothelial progenitor cell suspension(1×10~(10) cell/L) via the tail vein, 120 mg/kg resveratrol via gavage or endothelial progenitor cell suspension plus resveratrol. The administration in each group was taken once a day for 7 consecutive days.RESULTS AND CONCLUSION:(1) Compared with the model group, the levels of urea nitrogen, creatinine, urinary acetylglucosaminidase,urinary γ-L-glutamyl transpeptidase and urine protein decreased significantly in the resveratrol group, endothelial progenitor cell group(P <0.05) and dramatically in the combined treatment group(P < 0.01).(2) Compared with the model group, the levels of reactive oxygen species and malondialdehyde were reduced in the resveratrol group and endothelial progenitor cell group, whereas the activities of superoxide dismutase and glutathione peroxidase increased significantly(P < 0.05). Compared with the other groups, the combined treatment group had a greater improvement in the above-mentioned renal tissue indicators(P < 0.01).(3) The histopathological improvement of the renal tissue in the combined treatment group was markedly better than that in the endothelial progenitor cell group, resveratrol and model group. Under the fluorescence microscopy, the number of CM-Dil-labeled endothelial progenitor cells was significantly higher in the combined treatment group than the endothelial progenitor cell group(P < 0.05). To conclude, resveratrol combined with endothelial progenitor cell transplantation protects against renal injury induced by chronic cadmium exposure.
BACKGROUND: Studies have shown that resveratrol can effectively improve the survival rate of rat proximal tubular epithelial cells and inhibit cell apoptosis to protect against renal tubular epithelial cell injury. However, the specific mechanism of resveratrol in the treatment of renal injury in rats remains to be further clarified.OBJECTIVE: To investigate the protective effect and mechanism of resveratrol combined with endothelial progenitor cell transplantation on renal injury caused by chronic cadmium exposure in rats.METHODS: Rat endothelial progenitor cells provided by the Chinese Academy of Medical Sciences were resuscitated and cultured in vitro,and cell suspensions were then prepared. Endothelial progenitor cells were labeled with CM-Dil. Wistar rats purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. in China were randomly divided into control group, pure cadmium poisoning group(model group),resveratrol group, endothelial progenitor cell group and combined treatment group, with 15 rats in each group. In the control group,physiological saline was injected subcutaneously. In the other four groups, the rats were subcutaneously administered with 6 μmol/kg cadmium chloride once daily for 6 weeks to establish animal models of renal injury. After modeling, the rat models in the latter four groups were given 0.5 mL of L-DMEM via the tail vein, 0.5 mL of endothelial progenitor cell suspension(1×10~(10) cell/L) via the tail vein, 120 mg/kg resveratrol via gavage or endothelial progenitor cell suspension plus resveratrol. The administration in each group was taken once a day for 7 consecutive days.RESULTS AND CONCLUSION:(1) Compared with the model group, the levels of urea nitrogen, creatinine, urinary acetylglucosaminidase,urinary γ-L-glutamyl transpeptidase and urine protein decreased significantly in the resveratrol group, endothelial progenitor cell group(P <0.05) and dramatically in the combined treatment group(P < 0.01).(2) Compared with the model group, the levels of reactive oxygen species and malondialdehyde were reduced in the resveratrol group and endothelial progenitor cell group, whereas the activities of superoxide dismutase and glutathione peroxidase increased significantly(P < 0.05). Compared with the other groups, the combined treatment group had a greater improvement in the above-mentioned renal tissue indicators(P < 0.01).(3) The histopathological improvement of the renal tissue in the combined treatment group was markedly better than that in the endothelial progenitor cell group, resveratrol and model group. Under the fluorescence microscopy, the number of CM-Dil-labeled endothelial progenitor cells was significantly higher in the combined treatment group than the endothelial progenitor cell group(P < 0.05). To conclude, resveratrol combined with endothelial progenitor cell transplantation protects against renal injury induced by chronic cadmium exposure.
引文
[1]余娜,钟志勇,唐小江,等.镉引起肾脏毒性的细胞凋亡通路[J].生态毒理学报,2014,9(3):407-412.
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[9]Pangeni R,Sahni JK,Ali J,et al.Resveratrol:review on therapeutic potential and recent advances in drug delivery.Expert Opin Drug Deliv.2014;11(8):1285.
[10]李艳,张颖,冯锦红,等.白藜芦醇对高脂饮食诱导肾损伤小鼠肾组织单核细胞趋化蛋白1及转化生长因子β1表达的影响[J].中国血液净化,2017,16(7):482-487.
[11]单娟萍,董梁,董志超,等.白藜芦醇抑制顺铂诱导肾小管上皮细胞凋亡的体外研究[J].中华全科医学,2017,15(8):1334-1336.
[12]周磊,李海伦,郑东辉.白藜芦醇在肾缺血再灌注损伤中的研究进展[J].徐州医学院学报,2016,36(1):64-67.
[13]Lu C,Zhang J,Zhang D,et al.EPCs in vascular repair:how can we clear the hurdles between bench and bedside?Front Biosci.2014;19(1):34-48.
[14]Rurali E,Bassetti B,Perrucci GL,et al.BM ageing:Implication for cell therapy with EPCs.Mech Ageing Dev.2016;159:4-13.
[15]Chong MS,Ng WK,Chan JK.Concise review:endothelial progenitor cells in regenerative medicine:applications and challenges.Stem Cells Transl Med.2016;5(4):530.
[16]Sun K,Zheng Z,Ju X,et al.Combined transplantation of mesenchymal stem cells and endothelial progenitor cells for tissue engineering:a systematic review and meta-analysis.Stem Cell Res Ther.2016;7(1):151.
[17]Salmina AB1.Neuron-glia interactions as therapeutic targets in neurodegeneration.J Alzheimers Dis.2009;16(3):485-502.
[18]袁娟,赖燕,张云花,等.职业性镉中毒肾功能预后分析[J].工业卫生与职业病,2016,42(6):452-454.
[19]Wang HJ,Liang R,Fu LM,et al.Nutritional aspects ofβ-carotene and resveratrol antioxidant synergism in giant unilamellar vesicles.Food Funct.2014;5(7):1573-1578.
[20]Dave M,Attur M,Palmer G,et al.The antioxidant resveratrol protects against chondrocyte apoptosis via effects on mitochondrial polarization and ATP production.Arthritis Rheum.2008;58(9):2786-2797.
[21]Bellaver B,Souza DG,Souza DO,et al.Resveratrol increases antioxidant defenses and decreases proinflammatory cytokines in hippocampal astrocyte cultures from newborn,adult and aged Wistar rats.Toxicol In Vitro.2014;28(4):479-484.
[22]Fu B,Zhao J,Peng W,et al.Resveratrol rescues cadmium-induced mitochondrial injury by enhancing transcriptional regulation of PGC-1αand SOD2 via the Sirt3/FoxO3a pathway in TCMK-1 cells.Biochem Biophys Res Commun.2017;486(1):198-204.
[23]Lee PS,Poh KK.Endothelial progenitor cells in cardiovascular diseases.World J Stem Cells.2014;6(3):355-366.
[24]李清,冯麟.尿蛋白联合检测对2型糖尿病早期肾损害的诊断价值探讨[J].临床医药文献电子杂志,2018,5(27):163-164.
[25]胡鹏华,储虹,梁馨苓,等.尿蛋白在预测老年心脏术后患者急性肾损伤中的价值[J].中华老年医学杂志,2018,37(11):1190-1195.
[26]Soler MJ,Martínez-Estrada OM,Puig-MaríJM,et al.Circulating endothelial progenitor cells after kidney transplantation.Am JTransplant.2005;5(9):2154-2159.
[27]Patschan D,Patschan S,Müller GA.Endothelial progenitor cells in acute ischemic kidney injury:strategies for increasing the cells'renoprotective competence.Int J Nephrol.2011;2011:828369.
[28]Satarug S,Vesey DA,Gobe GC.Kidney cadmium toxicity,diabetes and high blood pressure:the perfect storm.Tohoku J Exp Med.2017;241(1):65-87.
[29]王庆华,顾建红,胥清芳等.铅和(或)镉对大鼠肾细胞的毒性损伤及硒的拮抗作用[J].中国兽医学报,2014,34(8):1353-1357.
[2]金媛媛,周蓉,匡兴亚.镉致肾损伤机制的研究进展[J].环境与职业医学,2018,35(2):180-184.
[3]李静慧,徐兆发.镉致肾细胞凋亡机制的研究进展及防治[J].毒理学杂志,2014,28(4):319-322.
[4]Nazima B,Manoharan V,Miltonprabu S.Grape seed proanthocyanidins ameliorates cadmium-induced renal injury and oxidative stress in experimental rats through the up-regulation of nuclear related factor 2 and antioxidant responsive elements.Biochem Cell Biol.2015;93(3):210-226.
[5]Hagar H,Al Malki W.Betaine supplementation protects against renal injury induced by cadmium intoxication in rats:role of oxidative stress and caspase-3.Environ Toxicol Pharmacol.2014;37(2):803-811.
[6]周庆彪,刘颖,孔德钦,等.硒对慢性镉暴露致大鼠肾损伤的保护作用与机制[J].癌变·畸变·突变,2018,30(1):1-7.
[7]Prozialeck WC,Lamar PC,Edwards JR.Effects of sub-chronic Cd exposure on levels of copper,selenium,zinc,iron and other essential metals in rat renal cortex.Toxicol Rep.2016;3:740-746.
[8]Park EJ,Pezzuto JM.The pharmacology of resveratrol in animals and humans.Mol Basis Dis.2015;1852(6):1071-1113.
[9]Pangeni R,Sahni JK,Ali J,et al.Resveratrol:review on therapeutic potential and recent advances in drug delivery.Expert Opin Drug Deliv.2014;11(8):1285.
[10]李艳,张颖,冯锦红,等.白藜芦醇对高脂饮食诱导肾损伤小鼠肾组织单核细胞趋化蛋白1及转化生长因子β1表达的影响[J].中国血液净化,2017,16(7):482-487.
[11]单娟萍,董梁,董志超,等.白藜芦醇抑制顺铂诱导肾小管上皮细胞凋亡的体外研究[J].中华全科医学,2017,15(8):1334-1336.
[12]周磊,李海伦,郑东辉.白藜芦醇在肾缺血再灌注损伤中的研究进展[J].徐州医学院学报,2016,36(1):64-67.
[13]Lu C,Zhang J,Zhang D,et al.EPCs in vascular repair:how can we clear the hurdles between bench and bedside?Front Biosci.2014;19(1):34-48.
[14]Rurali E,Bassetti B,Perrucci GL,et al.BM ageing:Implication for cell therapy with EPCs.Mech Ageing Dev.2016;159:4-13.
[15]Chong MS,Ng WK,Chan JK.Concise review:endothelial progenitor cells in regenerative medicine:applications and challenges.Stem Cells Transl Med.2016;5(4):530.
[16]Sun K,Zheng Z,Ju X,et al.Combined transplantation of mesenchymal stem cells and endothelial progenitor cells for tissue engineering:a systematic review and meta-analysis.Stem Cell Res Ther.2016;7(1):151.
[17]Salmina AB1.Neuron-glia interactions as therapeutic targets in neurodegeneration.J Alzheimers Dis.2009;16(3):485-502.
[18]袁娟,赖燕,张云花,等.职业性镉中毒肾功能预后分析[J].工业卫生与职业病,2016,42(6):452-454.
[19]Wang HJ,Liang R,Fu LM,et al.Nutritional aspects ofβ-carotene and resveratrol antioxidant synergism in giant unilamellar vesicles.Food Funct.2014;5(7):1573-1578.
[20]Dave M,Attur M,Palmer G,et al.The antioxidant resveratrol protects against chondrocyte apoptosis via effects on mitochondrial polarization and ATP production.Arthritis Rheum.2008;58(9):2786-2797.
[21]Bellaver B,Souza DG,Souza DO,et al.Resveratrol increases antioxidant defenses and decreases proinflammatory cytokines in hippocampal astrocyte cultures from newborn,adult and aged Wistar rats.Toxicol In Vitro.2014;28(4):479-484.
[22]Fu B,Zhao J,Peng W,et al.Resveratrol rescues cadmium-induced mitochondrial injury by enhancing transcriptional regulation of PGC-1αand SOD2 via the Sirt3/FoxO3a pathway in TCMK-1 cells.Biochem Biophys Res Commun.2017;486(1):198-204.
[23]Lee PS,Poh KK.Endothelial progenitor cells in cardiovascular diseases.World J Stem Cells.2014;6(3):355-366.
[24]李清,冯麟.尿蛋白联合检测对2型糖尿病早期肾损害的诊断价值探讨[J].临床医药文献电子杂志,2018,5(27):163-164.
[25]胡鹏华,储虹,梁馨苓,等.尿蛋白在预测老年心脏术后患者急性肾损伤中的价值[J].中华老年医学杂志,2018,37(11):1190-1195.
[26]Soler MJ,Martínez-Estrada OM,Puig-MaríJM,et al.Circulating endothelial progenitor cells after kidney transplantation.Am JTransplant.2005;5(9):2154-2159.
[27]Patschan D,Patschan S,Müller GA.Endothelial progenitor cells in acute ischemic kidney injury:strategies for increasing the cells'renoprotective competence.Int J Nephrol.2011;2011:828369.
[28]Satarug S,Vesey DA,Gobe GC.Kidney cadmium toxicity,diabetes and high blood pressure:the perfect storm.Tohoku J Exp Med.2017;241(1):65-87.
[29]王庆华,顾建红,胥清芳等.铅和(或)镉对大鼠肾细胞的毒性损伤及硒的拮抗作用[J].中国兽医学报,2014,34(8):1353-1357.