茵陈提取物对糖尿病大鼠肾组织中miRNAs表达谱的影响及其肾脏保护作用
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摘要
目的:观察茵陈提取物(HACE)对糖尿病大鼠肾组织中微小核糖核酸(miRNAs)表达谱的影响,从miRNA角度探讨HACE对糖尿病大鼠的肾脏保护作用及其机制。方法:采用链脲佐菌素(STZ)复制糖尿病大鼠模型,将60只雄性Wistar大鼠分为对照组(12只)、模型组(24只)和HACE组(24只)。分别于给药8和16周后检测各组大鼠尿白蛋白排泄率和尿蛋白排泄率;HE染色检测大鼠肾组织形态表现;miRNA芯片初步筛查各组大鼠肾组织中miRNAs差异表达谱,对于差异表达在1.74倍以上高丰度的miRNAs筛选阳性miRNAs,采用RT-qPCR验证筛选的miRNAs的相对表达量。结果:与模型组比较,给药8和16周后HACE组大鼠肾小球系膜聚集等现象明显减轻,尿白蛋白排泄率明显降低(P<0.05)。miRNA芯片初筛,对照组和模型组大鼠肾组织中有35个差异表达的miRNAs,模型组和HACE组大鼠肾组织中有17个差异表达的miRNAs。RT-qPCR法检测,给药8周后,与对照组比较,模型组大鼠肾组织中miR-1306-3p(P<0.05)、miR-672-5p(P<0.05)和miR-3550(P<0.01)相对表达量明显降低;与模型组比较,HACE组大鼠肾组织中miR-672-5p相对表达量升高(P<0.05)。给药16周后,与对照组比较,模型组大鼠肾组织中miR-21-5p相对表达量升高(P<0.01),miR-1306-3p(P<0.05)、miR-672-5p(P<0.01)和miR-466d(P<0.05)相对表达量降低;与模型组比较,HACE组大鼠肾组织中miR-21-5p(P<0.05)和miR-1306-3p(P<0.05)相对表达量降低,miR-672-5p相对表达量升高(P<0.05)。结论:HACE作用后糖尿病大鼠肾组织中miRNAs表达谱出现变化。HACE对糖尿病大鼠的肾脏具有保护作用,其机制可能与miRNAs有关。
Objective:To observe the effect of herba artemisiae capillaris extracts(HACE)on the expression profile of microRNA(miRNAs)in the kidney tissue of the diabetic rats,and to explore the protective effect of HACE on the kidney tissue of diabetic rats from the miRNA angle.Methods:A total of 60 male Wistar rats were divided into control group(n=12),model group(n=24)and HACE group(n=24).The urinary albumin excretion rate and urinary protein excretion rate of the rats in various groups were detected.HE staining was used to detect the morphology of the kidney tissue of the rats in various groups after treated for 8 and 16 weeks;miRNAs chips were used to screen the expressions of miRNAs in the kidney tissue of the rats.The positive miRNAs were screened out from those with differentially expression amounts higher than 1.74 times as well as high abundance,and RT-qPCR was used to verify the relative expression amounts of miRNAs.Results:Compared with model group,the glomeruli matrix accumulation of the rats in HACE group(after treated for 8 and 16 weeks)was significantly improved and the urinary albumin excretion rates were decreased(P<0.05).The miRNAs chips results showed that there were 35 differentially expressed miRNAs in control and model groups and there were17 differentially expressed miRNAs in model and HACE groups.The RT-qPCR results showed that after treated for8 weeks,compared with control group,the relative expression amounts of miR-1306-3 p(P<0.05),miR-672-5 p(P<0.05)and miR-3550(P<0.01)in the kidney tissue of the rats in model group were decreased;compared with model group,the relative expression amount of miR-672-5 p in the kidney tissue of the rats in HACE group was increased(P<0.05).After treated for 16 weeks,compared with control group,the relative expression amount of miR-21-5 p in the kidney tissue of the rats in model group was increased(P <0.01),the relative expression amounts of miR-1306-3 p(P <0.05),miR-672-5 p(P <0.01)and miR-466 d(P <0.05)were decreased;compared with model group,the relative expression amounts of miR-21-5 p(P<0.05)and miR-1306-3 p(P<0.05)in the kidney tissue of the rats in HACE group were decreased,and the relative expression amount of miR-672-5 p was increased(P<0.05).Conclusion:The miRNAs expression profile in kidney tissue of diabetic rats were changed after HACE treatment.HACE has the protectively effect on the kidney tissue of diabetic rats and the mechanism may be associated with miRNAs.
Objective:To observe the effect of herba artemisiae capillaris extracts(HACE)on the expression profile of microRNA(miRNAs)in the kidney tissue of the diabetic rats,and to explore the protective effect of HACE on the kidney tissue of diabetic rats from the miRNA angle.Methods:A total of 60 male Wistar rats were divided into control group(n=12),model group(n=24)and HACE group(n=24).The urinary albumin excretion rate and urinary protein excretion rate of the rats in various groups were detected.HE staining was used to detect the morphology of the kidney tissue of the rats in various groups after treated for 8 and 16 weeks;miRNAs chips were used to screen the expressions of miRNAs in the kidney tissue of the rats.The positive miRNAs were screened out from those with differentially expression amounts higher than 1.74 times as well as high abundance,and RT-qPCR was used to verify the relative expression amounts of miRNAs.Results:Compared with model group,the glomeruli matrix accumulation of the rats in HACE group(after treated for 8 and 16 weeks)was significantly improved and the urinary albumin excretion rates were decreased(P<0.05).The miRNAs chips results showed that there were 35 differentially expressed miRNAs in control and model groups and there were17 differentially expressed miRNAs in model and HACE groups.The RT-qPCR results showed that after treated for8 weeks,compared with control group,the relative expression amounts of miR-1306-3 p(P<0.05),miR-672-5 p(P<0.05)and miR-3550(P<0.01)in the kidney tissue of the rats in model group were decreased;compared with model group,the relative expression amount of miR-672-5 p in the kidney tissue of the rats in HACE group was increased(P<0.05).After treated for 16 weeks,compared with control group,the relative expression amount of miR-21-5 p in the kidney tissue of the rats in model group was increased(P <0.01),the relative expression amounts of miR-1306-3 p(P <0.05),miR-672-5 p(P <0.01)and miR-466 d(P <0.05)were decreased;compared with model group,the relative expression amounts of miR-21-5 p(P<0.05)and miR-1306-3 p(P<0.05)in the kidney tissue of the rats in HACE group were decreased,and the relative expression amount of miR-672-5 p was increased(P<0.05).Conclusion:The miRNAs expression profile in kidney tissue of diabetic rats were changed after HACE treatment.HACE has the protectively effect on the kidney tissue of diabetic rats and the mechanism may be associated with miRNAs.
引文
[1]Vaca L.Point-of-care diagnostic tools to detect circulating microRNAS as biomarkers of disease[J].Sensors(Basel),2014,14(5):9117-9131.
[2]Wu H,Kong L,Zhou S,et al.The role of microRNAs in diabetic nephropathy[J].J Diabetes Res,2014,2014:920134.
[3]Castro NE,Kato M,Park JT,et al.Transforming growth factorβ1(TGF-β1)enhances expression of profibrotic genes through a novel signaling cascade and microRNAs in renal mesangial cells[J].J Biol Chem,2014,289(42):29001-29013.
[4]Long J,Wang Y,Wang W,et al.MicroRNA-29cis a signature microRNA under high glucose conditions that targets Sprouty homolog 1,and its in vivo knockdown prevents progression of diabetic nephropathy[J].J Biol Chem,2011,286(13):11837-11848.
[5]Kato M,Zhang J,Wang M,et al.MicroRNA-192 in diabetic kidney glomeruli and its function in TGF-beta-induced collagen expression via inhibition of E-box repressors[J].Proc Natl Acad Sci U S A,2007,104(9):3432-3437.
[6]Wang Q,Wang Y,Minto AW,et al.MicroRNA-377is upregulated and can lead to increased fibronectin production in diabetic nephropathy[J].FASEB J,2008,22(12):4126-4135.
[7]Zhao B,Li H,Liu J,et al.MicroRNA-23btargets ras GTPase-activating protein SH3 domain-binding protein 2to alleviate fibrosis and albuminuria in diabetic nephropathy[J].J Am Chem Soc,2016,27(9):2597-2608.
[8]Koga K,Yokoi H,Mori K,et al.MicroRNA-26ainhibits TGF-β-induced extracellular matrix protein expression in podocytes by targeting CTGF and is downregulated in diabetic nephropathy[J].Diabetologia,2015,58(9):2169-2180.
[9]Wang J,Duan L,Guo T,et al.Downregulation of miRNA-30c promotes renal fibrosis by target CTGF in diabetic nephropathy[J].J Diabetes Complicat,2016,30(3):406-414.
[10]Etikala DM,Liu R,Wang L.FOXP3-micro RNA-146-NFkappa B as oncotarget[J].Oncoscience,2015,2(10):839-840.
[11]Wang X,Shen E,Wang Y,et al.MiR-196aregulates high glucose-induced mesangial cell hypertrophy by targeting p27kip1[J].J Lab Autom,2015,20(4):491-499.
[12]Zhang Z,Luo X,Ding S,et al.MicroRNA-451regulates p38 MAPK signaling by targeting of Ywhaz and suppresses the mesangial hypertrophy in early diabetic nephropathy[J].FEBS Lett,2012,586(1):20-26.
[13]Baker MA,Davis SJ,Liu P,et al.Tissue-specific microRNA expression patterns in four types of kidney disease[J].J Am Soc Nephrol,2017,28(10):2985-2992.
[14]潘竞锵,刘广南.茵陈蒿对小鼠血糖、血脂的影响[J].中药材,1998,21(8):408-411.
[15]潘竞锵,韩超,刘惠纯,等.茵陈蒿汤对正常和多种糖尿病模型动物血糖的影响[J].中药材,2001,24(2):128-131.
[16]栾智杰.茵陈蒿汤对腺嘌呤肾衰大鼠CTGF和TGF-β1表达的影响[D].哈尔滨:黑龙江省中医研究院,2011.
[17]耿嘉男.茵陈提取物对糖尿病肾病防治作用的研究[D].长春:吉林大学,2015.
[18]彭睿,查何,周吉,等.糖尿病肾病循环microRNA差异表达谱的分析[J].西南大学学报:自然科学版,2014,36(11):75-82.
[19]张政,罗勇军,彭惠民,等.早期糖尿病肾病相关微小RNA在小鼠肾脏的表达变化[J].第三军医大学学报,2010,32(13):1383-1386.
[20]杨雪唯,梁小弟,努尔斯曼古丽·奥斯曼,等.MicroRNA及其靶基因在糖尿病肾病小鼠肾脏的表达[J].科技导报,2017,35(4):84-89.
[21]Campion CG,Sanchez-Ferras O,Batchu SN.Potential role of serum and urinary biomarkers in diagnosis and prognosis of diabetic nephropathy[J].Can J Kidney Health Dis,2017,22(4):205435811770537.
[22]郝传明,张敏.糖尿病肾病的防治策略—三级预防一体化治疗[J].中国实用内科杂志,2017,37(3):195-198.
[23]孙赟.复方积雪草合剂治疗早期糖尿病肾病临床疗效观察[J].中国实用内科杂志,2017,37(5):467-468.
[2]Wu H,Kong L,Zhou S,et al.The role of microRNAs in diabetic nephropathy[J].J Diabetes Res,2014,2014:920134.
[3]Castro NE,Kato M,Park JT,et al.Transforming growth factorβ1(TGF-β1)enhances expression of profibrotic genes through a novel signaling cascade and microRNAs in renal mesangial cells[J].J Biol Chem,2014,289(42):29001-29013.
[4]Long J,Wang Y,Wang W,et al.MicroRNA-29cis a signature microRNA under high glucose conditions that targets Sprouty homolog 1,and its in vivo knockdown prevents progression of diabetic nephropathy[J].J Biol Chem,2011,286(13):11837-11848.
[5]Kato M,Zhang J,Wang M,et al.MicroRNA-192 in diabetic kidney glomeruli and its function in TGF-beta-induced collagen expression via inhibition of E-box repressors[J].Proc Natl Acad Sci U S A,2007,104(9):3432-3437.
[6]Wang Q,Wang Y,Minto AW,et al.MicroRNA-377is upregulated and can lead to increased fibronectin production in diabetic nephropathy[J].FASEB J,2008,22(12):4126-4135.
[7]Zhao B,Li H,Liu J,et al.MicroRNA-23btargets ras GTPase-activating protein SH3 domain-binding protein 2to alleviate fibrosis and albuminuria in diabetic nephropathy[J].J Am Chem Soc,2016,27(9):2597-2608.
[8]Koga K,Yokoi H,Mori K,et al.MicroRNA-26ainhibits TGF-β-induced extracellular matrix protein expression in podocytes by targeting CTGF and is downregulated in diabetic nephropathy[J].Diabetologia,2015,58(9):2169-2180.
[9]Wang J,Duan L,Guo T,et al.Downregulation of miRNA-30c promotes renal fibrosis by target CTGF in diabetic nephropathy[J].J Diabetes Complicat,2016,30(3):406-414.
[10]Etikala DM,Liu R,Wang L.FOXP3-micro RNA-146-NFkappa B as oncotarget[J].Oncoscience,2015,2(10):839-840.
[11]Wang X,Shen E,Wang Y,et al.MiR-196aregulates high glucose-induced mesangial cell hypertrophy by targeting p27kip1[J].J Lab Autom,2015,20(4):491-499.
[12]Zhang Z,Luo X,Ding S,et al.MicroRNA-451regulates p38 MAPK signaling by targeting of Ywhaz and suppresses the mesangial hypertrophy in early diabetic nephropathy[J].FEBS Lett,2012,586(1):20-26.
[13]Baker MA,Davis SJ,Liu P,et al.Tissue-specific microRNA expression patterns in four types of kidney disease[J].J Am Soc Nephrol,2017,28(10):2985-2992.
[14]潘竞锵,刘广南.茵陈蒿对小鼠血糖、血脂的影响[J].中药材,1998,21(8):408-411.
[15]潘竞锵,韩超,刘惠纯,等.茵陈蒿汤对正常和多种糖尿病模型动物血糖的影响[J].中药材,2001,24(2):128-131.
[16]栾智杰.茵陈蒿汤对腺嘌呤肾衰大鼠CTGF和TGF-β1表达的影响[D].哈尔滨:黑龙江省中医研究院,2011.
[17]耿嘉男.茵陈提取物对糖尿病肾病防治作用的研究[D].长春:吉林大学,2015.
[18]彭睿,查何,周吉,等.糖尿病肾病循环microRNA差异表达谱的分析[J].西南大学学报:自然科学版,2014,36(11):75-82.
[19]张政,罗勇军,彭惠民,等.早期糖尿病肾病相关微小RNA在小鼠肾脏的表达变化[J].第三军医大学学报,2010,32(13):1383-1386.
[20]杨雪唯,梁小弟,努尔斯曼古丽·奥斯曼,等.MicroRNA及其靶基因在糖尿病肾病小鼠肾脏的表达[J].科技导报,2017,35(4):84-89.
[21]Campion CG,Sanchez-Ferras O,Batchu SN.Potential role of serum and urinary biomarkers in diagnosis and prognosis of diabetic nephropathy[J].Can J Kidney Health Dis,2017,22(4):205435811770537.
[22]郝传明,张敏.糖尿病肾病的防治策略—三级预防一体化治疗[J].中国实用内科杂志,2017,37(3):195-198.
[23]孙赟.复方积雪草合剂治疗早期糖尿病肾病临床疗效观察[J].中国实用内科杂志,2017,37(5):467-468.