茵陈提取物对糖尿病大鼠肾组织中PTEN蛋白表达的影响及其肾脏保护作用
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摘要
目的:探讨茵陈提取物(HACE)对糖尿病大鼠肾组织中第10号染色体缺失的磷酸酶及张力蛋白同源基因(PTEN)表达的影响,阐明HACE对糖尿病大鼠肾脏保护作用的药理学机制。方法:采用链脲佐菌素(STZ)制备糖尿病大鼠模型。30只Wistar大鼠随机分为对照组(6只)、模型组(12只)和HACE组(12只),HACE组大鼠每天1次灌胃给予HACE (5g·kg-1),对照组和模型组大鼠每天1次灌胃给予等量生理盐水。给药4个月后检测各组大鼠尿白蛋白排泄率和尿总蛋白排泄率,HE染色观察大鼠肾组织病理形态表现,PAS和Masson染色检测肾小球细胞外基质(ECM)分布情况,免疫组织化学染色法检测大鼠肾组织中PTEN蛋白表达分布情况,Western blotting法检测各组大鼠肾脏组织中PTEN蛋白表达水平。结果:与模型组比较,HACE组大鼠肾小球系膜区扩大、PAS及Masson阳性染色物质聚集和基底膜增厚等现象明显减轻,24h尿白蛋白排泄率明显降低(P<0.05),尿总蛋白排泄率差异无统计学意义(P>0.05)。免疫组织化学染色检测,对照组大鼠肾组织PTEN蛋白表达量较高,模型组大鼠肾组织中PTEN蛋白表达量降低,HACE组大鼠肾组织中PTEN蛋白表达量趋向正常。Western blotting法检测,与对照组比较,模型组大鼠肾皮质中PTEN蛋白表达水平明显降低(P<0.05);与模型组比较,HACE组大鼠肾皮质组织中PTEN蛋白表达水平明显升高(F=5.06,P<0.05)。结论:HACE提高大鼠肾脏组织中PTEN蛋白的表达可能是其对糖尿病大鼠肾脏保护作用机制之一。
Objective:To investigate the effect of herba artemisiae capillaris extracts(HACE)on the expression of phosphatase and tensin homologue deleted chromatosome 10(PTEN)in the kidney tissue of the diabetic rats,and to clarify the pharmacological mechanism of protective effect of HACE on the kidney of the diabetic rats.Methods:Thirty Wistar rats were randomly divided into control group(n=6),model group(n=12)and HACE group(n=12).The rats in HACE group were intragastrically administrated with HACE(5 g·kg-1)one time every day,and the rats in control group and model group were intragastrically administrated with the same volume of normal saline.The diabetic rat models were duplicated by streptozotocin(STZ). After 4 months of administration,the urinary albumin excretion rates and urinary total protein excretion rates of the rats in various groups were detected.The pathomorphology of the kindey tissue of the rats in various groups were observed by HE staining and the distribution of renal extracellular matrix(ECM)was detected by PAS and Masson staining.The expressions of PTEN protein in kidney tissue of the rats in various groups were detected by immunohistostaining and the expression levels of PTEN protein in kidney tissue of the rats in various groups were detected by Western blotting method.Results:Compared with model group,the glomerular mesangial aggregation,positive staining of PAS as well as Masson aggregation and basement membrane thickening of the rats in HACE group were significantly reduced,and the 24 hurinary albumin excretion rate was significantly decreased(P<0.05),and the urinary total protein excretion rate had no significant difference(P>0.05).The immunohistochemistry staining results showed that the expression level of PTEN protein in kidney tissue of the rats in control group was higher,the expression level of PTEN protein in the kidney tissue of the rats in model group was decreased,and the expression level of PTEN protein in kidney tissue of the rats in HACE group was increased to normal.The Western blotting results showed that the expression level of PTEN protein in the renal cortex tissue tissue of the rats in model group was significantly decreased compared with control group(P<0.05);compared with model group,the expression level of PTEN protein in renal cortex tissue of the rats in HACE group was significantly increased(P<0.05).Conclusion:HACE can increase the expression of PTEN protein in kidney tissue of the rats,which may be one of the mechanisms of its protective effect on the kidney in the diabetic rats.
Objective:To investigate the effect of herba artemisiae capillaris extracts(HACE)on the expression of phosphatase and tensin homologue deleted chromatosome 10(PTEN)in the kidney tissue of the diabetic rats,and to clarify the pharmacological mechanism of protective effect of HACE on the kidney of the diabetic rats.Methods:Thirty Wistar rats were randomly divided into control group(n=6),model group(n=12)and HACE group(n=12).The rats in HACE group were intragastrically administrated with HACE(5 g·kg-1)one time every day,and the rats in control group and model group were intragastrically administrated with the same volume of normal saline.The diabetic rat models were duplicated by streptozotocin(STZ). After 4 months of administration,the urinary albumin excretion rates and urinary total protein excretion rates of the rats in various groups were detected.The pathomorphology of the kindey tissue of the rats in various groups were observed by HE staining and the distribution of renal extracellular matrix(ECM)was detected by PAS and Masson staining.The expressions of PTEN protein in kidney tissue of the rats in various groups were detected by immunohistostaining and the expression levels of PTEN protein in kidney tissue of the rats in various groups were detected by Western blotting method.Results:Compared with model group,the glomerular mesangial aggregation,positive staining of PAS as well as Masson aggregation and basement membrane thickening of the rats in HACE group were significantly reduced,and the 24 hurinary albumin excretion rate was significantly decreased(P<0.05),and the urinary total protein excretion rate had no significant difference(P>0.05).The immunohistochemistry staining results showed that the expression level of PTEN protein in kidney tissue of the rats in control group was higher,the expression level of PTEN protein in the kidney tissue of the rats in model group was decreased,and the expression level of PTEN protein in kidney tissue of the rats in HACE group was increased to normal.The Western blotting results showed that the expression level of PTEN protein in the renal cortex tissue tissue of the rats in model group was significantly decreased compared with control group(P<0.05);compared with model group,the expression level of PTEN protein in renal cortex tissue of the rats in HACE group was significantly increased(P<0.05).Conclusion:HACE can increase the expression of PTEN protein in kidney tissue of the rats,which may be one of the mechanisms of its protective effect on the kidney in the diabetic rats.
引文
[1]潘竞锵,刘广南,刘惠纯,等.茵陈蒿对小鼠血糖、血脂的影响[J].中药材,1998,21(8):408-411.
[2]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.
[3]袁效涵,石鹤峰,韩伟峰.加味茵陈蒿汤治疗2型糖尿病40例[J].中医研究,2007,20(2):43-44.
[4]GENG J N,YU X Y,LIU C Y,et al.Herba artemisiae capillaris extract prevents the development of streptozotocininduced diabetic nephropathy of rat[J].Evid Based Complement Alternat Med,2018,2018:5180165.
[5]孙成博,孙波,刘春禹,等.茵陈提取物对糖尿病大鼠肾组织miRNAs表达谱的影响及其肾脏保护作用[J].吉林大学学报:医学版,2018,44(3):493-498.
[6]MAEHAMA T,DIXON J E.The tumor suppressor,PTEN/MMAC1,dephosphorylates the lipid second messenger,phosphatidylinositol 3,4,5-trisphosphate[J].J Biol Chem,1998,273(22):13375-13378.
[7]刘伦华,楼丽广.PTEN功能调节的研究进展[J].中国药理学通报,2005,21(7):778-781.
[8]HADDADI N,LIN Y,TRAVIS G,et al.PTEN/PTENP1:Regulating the regulator of RTK-dependent PI3K/Akt signalling,new targets for cancer therapy[J].Mol Cancer,2018,17(1):37.
[9]LEE Y J,HAN H J.Troglitazone ameliorates high glucoseinduced EMT and dysfunction of SGLTs through PI3K/Akt,GSK-3β,Snail1,andβ-catenin in renal proximal tubule cells[J].Am J Physiol Renal Physiol,2010,29 8(5):F1263-F1275.
[10]HAO J,LIU S,ZHAO S,et al.PI3K/Akt pathway mediates high glucose-induced lipogenesis and extracellular matrix accumulation in HKC cells through regulation of SREBP-1 and TGF-β1[J].Histochem Cell Biol,2011,135(2):173-181.
[11]WANG X M,YAO M,LIU S X,et al.Interplay between the Notch and PI3K/Akt pathways in high glucose-induced podocyte apoptosis[J].Am J Physiol Renal Physiol,2014,306(2):F205-F213.
[12]LI X Y,WANG S S,HAN Z,et al.Triptolide restores autophagy to alleviate diabetic renal fibrosis through the miR-141-3p/PTEN/Akt/mTOR pathway[J].Mol Ther Nucleic Acids,2017,9:48-56.
[13]WANG H,FENG Z,XIE J,et al.Podocyte-specific knockin of PTEN protects kidney from hyperglycemia[J].Am J Physiol Renal Physiol,2018,314(6):F1096-F1107.
[14]SUN J,LI Z P,ZHANG R Q,et al.Repression of miR-217protects against high glucose-induced podocyte injury and insulin resistance by restoring PTEN-mediated autophagy pathway[J].Biochem Biophys Res Commun,2017,483(1):318-324.
[15]邢玲玲,傅淑霞,杨林,等.PTEN在糖尿病肾病患者足细胞损伤中的作用[J].临床与实验病理学杂志,2014,30(12):1375-1378.
[16]MAHIMAINATHAN L,DAS F,VENKATESAN B,et al.Mesangial cell hypertrophy by high glucose is mediated by downregulation of the tumor suppressor PTEN[J].Diabetes,2006,55(7):2115-2125.
[17]CAMPION C G,SANCHEZ-FERRAS O,BATCHU S N.Potential role of serum and urinary biomarkers in diagnosis and prognosis of diabetic nephropathy[J].Can J Kidney Health Dis,2017,4:2054358117705371.
[18]王圆圆,刘瑞霞,郭兵,等.大鼠肾组织PETN表达下调在糖尿病肾病发展中的作用[J].生理学报,2011,63(4):325-332.
[19]程悦,王代红,张耀全,等.PTEN编码蛋白对TGF-β1刺激大鼠肾成纤维细胞分泌ColⅣ、FN的抑制作用[J].解放军医学杂志,2007,32(7):719-721.
[20]HLOBILKOVA,KNILLOVJ,BRTEK J,et al.The mechanism of action of the tumour suppressor gene PTEN[J].Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub,2003,147(1):19-25.
[2]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.
[3]袁效涵,石鹤峰,韩伟峰.加味茵陈蒿汤治疗2型糖尿病40例[J].中医研究,2007,20(2):43-44.
[4]GENG J N,YU X Y,LIU C Y,et al.Herba artemisiae capillaris extract prevents the development of streptozotocininduced diabetic nephropathy of rat[J].Evid Based Complement Alternat Med,2018,2018:5180165.
[5]孙成博,孙波,刘春禹,等.茵陈提取物对糖尿病大鼠肾组织miRNAs表达谱的影响及其肾脏保护作用[J].吉林大学学报:医学版,2018,44(3):493-498.
[6]MAEHAMA T,DIXON J E.The tumor suppressor,PTEN/MMAC1,dephosphorylates the lipid second messenger,phosphatidylinositol 3,4,5-trisphosphate[J].J Biol Chem,1998,273(22):13375-13378.
[7]刘伦华,楼丽广.PTEN功能调节的研究进展[J].中国药理学通报,2005,21(7):778-781.
[8]HADDADI N,LIN Y,TRAVIS G,et al.PTEN/PTENP1:Regulating the regulator of RTK-dependent PI3K/Akt signalling,new targets for cancer therapy[J].Mol Cancer,2018,17(1):37.
[9]LEE Y J,HAN H J.Troglitazone ameliorates high glucoseinduced EMT and dysfunction of SGLTs through PI3K/Akt,GSK-3β,Snail1,andβ-catenin in renal proximal tubule cells[J].Am J Physiol Renal Physiol,2010,29 8(5):F1263-F1275.
[10]HAO J,LIU S,ZHAO S,et al.PI3K/Akt pathway mediates high glucose-induced lipogenesis and extracellular matrix accumulation in HKC cells through regulation of SREBP-1 and TGF-β1[J].Histochem Cell Biol,2011,135(2):173-181.
[11]WANG X M,YAO M,LIU S X,et al.Interplay between the Notch and PI3K/Akt pathways in high glucose-induced podocyte apoptosis[J].Am J Physiol Renal Physiol,2014,306(2):F205-F213.
[12]LI X Y,WANG S S,HAN Z,et al.Triptolide restores autophagy to alleviate diabetic renal fibrosis through the miR-141-3p/PTEN/Akt/mTOR pathway[J].Mol Ther Nucleic Acids,2017,9:48-56.
[13]WANG H,FENG Z,XIE J,et al.Podocyte-specific knockin of PTEN protects kidney from hyperglycemia[J].Am J Physiol Renal Physiol,2018,314(6):F1096-F1107.
[14]SUN J,LI Z P,ZHANG R Q,et al.Repression of miR-217protects against high glucose-induced podocyte injury and insulin resistance by restoring PTEN-mediated autophagy pathway[J].Biochem Biophys Res Commun,2017,483(1):318-324.
[15]邢玲玲,傅淑霞,杨林,等.PTEN在糖尿病肾病患者足细胞损伤中的作用[J].临床与实验病理学杂志,2014,30(12):1375-1378.
[16]MAHIMAINATHAN L,DAS F,VENKATESAN B,et al.Mesangial cell hypertrophy by high glucose is mediated by downregulation of the tumor suppressor PTEN[J].Diabetes,2006,55(7):2115-2125.
[17]CAMPION C G,SANCHEZ-FERRAS O,BATCHU S N.Potential role of serum and urinary biomarkers in diagnosis and prognosis of diabetic nephropathy[J].Can J Kidney Health Dis,2017,4:2054358117705371.
[18]王圆圆,刘瑞霞,郭兵,等.大鼠肾组织PETN表达下调在糖尿病肾病发展中的作用[J].生理学报,2011,63(4):325-332.
[19]程悦,王代红,张耀全,等.PTEN编码蛋白对TGF-β1刺激大鼠肾成纤维细胞分泌ColⅣ、FN的抑制作用[J].解放军医学杂志,2007,32(7):719-721.
[20]HLOBILKOVA,KNILLOVJ,BRTEK J,et al.The mechanism of action of the tumour suppressor gene PTEN[J].Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub,2003,147(1):19-25.
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