三七颗粒对STZ诱导的糖尿病模型大鼠足细胞转分化的影响
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摘要
目的:研究三七颗粒对链脲佐菌素(STZ)诱导的1型糖尿病模型大鼠的肾脏保护作用和足细胞上皮-间充质转分化(EMT)的影响.方法:24只8周龄雄性大鼠,随机选取8只为对照组,其余大鼠1次性腹腔注射质量分数为55 mg/kg的STZ构建1型糖尿病大鼠模型.随后,将糖尿病模型大鼠随机分为模型组和三七颗粒组(8只/组).三七颗粒组大鼠每天给予质量分数为0.4 g/kg的三七颗粒水溶液灌胃,对照组、模型组给予等量蒸馏水灌胃,连续12周.灌胃12周末测定大鼠血糖,检测尿微量白蛋白、尿肌酐、血肌酐及肾脏病理损伤,采用免疫组化法检测肾组织足细胞转分化骨架蛋白(desmin)、肾病蛋白(nephrin)和α平滑肌动蛋白(α-SMA)的表达.结果:与对照组比较,造模72 h后模型大鼠血糖浓度≥16.7 mmol/L,表明1型糖尿病大鼠模型建立成功.与模型组比较,三七颗粒组大鼠的尿微量白蛋白/尿肌酐的值显著降低(P<0.05);肾脏系膜增生面积率明显减少,肾脏足细胞足突融合数量明显减少(P<0.05);肾组织中的desmin蛋白和α-SMA蛋白的表达显著降低(P<0.05);nephrin蛋白的表达显著升高(P<0.05).结论:三七颗粒可改善1型糖尿病模型大鼠足细胞的转分化程度,缓解肾脏病理损伤,降低尿微量白蛋白/尿肌酐的值,可能因此而延缓糖尿病肾病的进展.
Objective:To study the renal protection of the Panax Notoginseng Granule-and its effect on podocyte epithelial-mesenchymal transition(EMT)in a streptozotocin(STZ)-induced type 1 diabetic rat model. Methods: Twenty-four male rats aged 8 weeks were used in this study. Eight rats were randomly selected as the control group. The rest of the rats were induced by a single intraperitoneal injection of STZ(55 mg/kg) for diabetic modeling. Then the diabetic rats were randomly divided into model group and Panax notoginseng granule group(n=8). Panax notoginseng granules group was given 0.4 g/kg aqueous solution by gavage every day, control group and modle group were given the same volume distilled water by gavage. The treatment lasted for 12 weeks, then fasting blood glucose(BG), 24 h Urinary albumin, Urinary creatinine, serum creatinine(Cr) and renal histopathology were examined. Immunohistochemistry was used to detect the protein expressions of desmin, nephrin and α-SMA. Results: Compared with the control group, 72 h after STZ injection, blood glucose of rats injected were higher than 16.7 mmol/L, indicating the model was established. Compared with the model group, Panax notoginseng granule group significantly decreased in the urinary albumin/urinary creatinine ratio(P<0.05), the area of mesangial hyperplasia in the kidney, the number of podocyte foot processes in the kidney(P<0.05), and the protein expression of desmin and α-SMA, whereas the protein expression of nephrin was up-regulated in Panax notoginseng granule group(P<0.05). Conclusion:Panax notoginseng granule can ameliorate podocyte EMT in STZ-induced type 1 diabetic rats, alleviate the pathological injury of the kidney, and reduce urinary albumin/creatinine ratio, so as to improve the progression of diabetic nephropathy.
Objective:To study the renal protection of the Panax Notoginseng Granule-and its effect on podocyte epithelial-mesenchymal transition(EMT)in a streptozotocin(STZ)-induced type 1 diabetic rat model. Methods: Twenty-four male rats aged 8 weeks were used in this study. Eight rats were randomly selected as the control group. The rest of the rats were induced by a single intraperitoneal injection of STZ(55 mg/kg) for diabetic modeling. Then the diabetic rats were randomly divided into model group and Panax notoginseng granule group(n=8). Panax notoginseng granules group was given 0.4 g/kg aqueous solution by gavage every day, control group and modle group were given the same volume distilled water by gavage. The treatment lasted for 12 weeks, then fasting blood glucose(BG), 24 h Urinary albumin, Urinary creatinine, serum creatinine(Cr) and renal histopathology were examined. Immunohistochemistry was used to detect the protein expressions of desmin, nephrin and α-SMA. Results: Compared with the control group, 72 h after STZ injection, blood glucose of rats injected were higher than 16.7 mmol/L, indicating the model was established. Compared with the model group, Panax notoginseng granule group significantly decreased in the urinary albumin/urinary creatinine ratio(P<0.05), the area of mesangial hyperplasia in the kidney, the number of podocyte foot processes in the kidney(P<0.05), and the protein expression of desmin and α-SMA, whereas the protein expression of nephrin was up-regulated in Panax notoginseng granule group(P<0.05). Conclusion:Panax notoginseng granule can ameliorate podocyte EMT in STZ-induced type 1 diabetic rats, alleviate the pathological injury of the kidney, and reduce urinary albumin/creatinine ratio, so as to improve the progression of diabetic nephropathy.
引文
[1] VARMA V K,KAJDACSY-BALLA A,AKKINA S K,et al.A label-free approach by infrared spectroscopic imaging for interrogating the biochemistry of diabetic nephropathy progression[J].Kidney Int,2016,89(5):1153-1159.
[2] OLATUNJI O J,CHEN H X,ZHOU Y F.Lycium chinense leaves extract ameliorates diabetic nephropathy by suppressing hyperglycemia mediated renal oxidative stress and inflammation[J].Biomed Pharmacother,2018,102:1145-1151.
[3] ZHANG X L,HE H,LIANG D,et al.Protective effects of berberine on renal injury in streptozotocin (STZ)-induced diabetic mice[J].Int J Mol Sci,2016,17(8):E1327.
[4] SAULNIER-BLACHE J S,FEIGERLOVA E,HALIMI J M,et al.Urinary lysophopholipids are increased in diabetic patients with nephropathy[J].J Diabetes Complications,2017,31(7):1103-1108.
[5] ZHAO L,WANG X L,SUN L N,et al.Critical role of serum response factor in podocyte epithelial-mesenchymal transition of diabetic nephropathy[J].Diab Vasc Dis Res,2016,13(1):81-92.
[6] CHEN X W,DU X Y,WANG Y X,et al.Irbesartan ameliorates diabetic nephropathy by suppressing the RANKL-RANK-NF-κB pathway in type 2 diabetic db/db mice[J].Mediators Inflamm,2016,2016:1405924.
[7] LING L,CHEN L B,ZHANG C N,et al.High glucose induces podocyte epithelial-to-mesenchymal transition by demethylation-mediated enhancement of MMP9 expression[J].Mol Med Rep,2018,17(4):5642-5651.
[8] YING Q D,WU G Z.Molecular mechanisms involved in podocyte EMT and concomitant diabetic kidney diseases:an update[J].Ren Fail,2017,39(1):474-483.
[9] LIU Z H.Nephrology in China[J].Nat Rev Nephrol,2013,9(9):523-528.
[10] LI B Q,CHEN J,WU T X,et al.Fast determination of four active compounds in Sanqi Panax Notoginseng Injection samples by high-performance liquid chromatography with a chemometric method[J].J Sep Sci,2015,38(9):1449-1457.
[11] 王巧凡.银可络联合三七、黄芪颗粒治疗糖尿病肾病疗效观察[J].中国中医急症,2007,16(3):294,296.WANG Q F.Efficacy of Yinkeluo combined with panax notoginseng and Huangqi granules in the treatment of diabetic nephropathy[J].Journal of Emergency in Traditional Chinese Medicine,2007,16(3):294,296.
[12] CENTER FOR DRUG EVALUATION AND RESEARCH (CDER).Estimating the maximum safe starting dose in initial clinical for therapeutics in adult healthy volunteers[S/OL].[2019-01-20] https://www.fda.gov/regulatory-information/search-fda-guidance-documents/estimating-maximum-safe-starting-dose-initial-clinical-trials-therapeutics-adult-healthy-volunteers.
[13] GUI D K,WEI L,JIAN G H,et al.Notoginsenoside R1 ameliorates podocyte adhesion under diabetic condition through α3β1 integrin upregulation in vitro and in vivo[J].Cell Physiol Biochem,2014,34(6):1849-1862.
[14] 肖磊,王华军,郑小飞,等.糖尿病对骨科手术的不利影响[J].暨南大学学报(自然科学与医学版),2018,39(5):426-434.XIAO L,WANG H J,ZHENG X F,et al.Overview of the pathology and adverse impacts of diabetes on orthopedic surgery[J].Journal of Jinan University(Natural Science & Medicine Edition),2018,39(5):426-434.
[15] 朱炳铭,吴勇,余权,等.血清miR-21联合尿微量白蛋白、尿微量白蛋白与尿肌酐比值诊断糖尿病肾病的价值[J].暨南大学学报(自然科学与医学版),2016,37(4):318-323.ZHU B M,WU Y,YU Q,et al.Clinical values of serum miR-21(SmiR-21) combined with urinary micro-albumin (UmAlb) and UmAlb /urine creatinine (UmAlb/Ucr) for diagnosis of diabetic nephropathy[J].Journal of Jinan University(Natural Science & Medicine Edition),2016,37(4):318-323.
[16] STINGHEN A E,MASSY Z A,VLASSARA H,et al.Uremic toxicity of advanced glycation end products in CKD[J].J Am Soc Nephrol,2016,27(2):354-370.
[17] 占永力,岳玉和,周静媛,等.中药三七注射液对显性糖尿病肾病并高凝状态的影响[J].中国中西医结合肾病杂志,2002,3(1):23-25.ZHAN Y L,YUE Y H,ZHOU J Y,et al.Effect of Xue Shuan Tong injection on overt diabetic nephropathy with hypercoagulability[J].Chinese Journal of Integrated Traditional and Western Nephrology,2002,3(1):23-25.
[18] 郑丽阳,薛瑞,吕亭亭,等.三七皂苷Fc对2型糖尿病db/db小鼠的肾脏保护作用研究[J].中华中医药学刊,2017,35(3):609-611.ZHENG L Y,XUE R,Lü T T,et al.Study on renal protective effect of notoginsenoside fc on db/db mice[J].Chinese Archives of Traditional Chinese Medicine,2017,35(3):609-611.
[19] HUANG G D,LV J,LI T Y,et al.Notoginsenoside R1 ameliorates podocyte injury in rats with diabetic nephropathy by activating the PI3K/Akt signaling pathway[J].Int J Mol Med,2016,38(4):1179-1189.
[20] 王洪敏,钟志勇,吴庆洲,等.大鼠糖尿病模型早期病理观察指标的筛选[J].广东药学院学报,2011,27(1):86-90.WANG H M,ZHONG Z Y,WU Q Z,et al.Selection of pathogenic indicators in the early stage of diabetes rats[J].Journal of Guangdong Pharmaceutical College,2011,27(1):86-90.
[21] MUNDEL P.Podocytes and the quest for precision medicines for kidney diseases[J].Pflugers Arch,2017,469(7/8):1029-1037.
[22] KANG Y S,LI Y J,DAI C S,et al.Inhibition of integrin-linked kinase blocks podocyte epithelial-mesenchymal transition and ameliorates proteinuria[J].Kidney Int,2010,78(4):363-373.
[23] DAI H R,LIU Q Q,LIU B L.Research progress on mechanism of podocyte depletion in diabetic nephropathy[J].J Diabetes Res,2017,2017:2615286.
[24] 王竹,刘俊田,孙万森,等.祛风通络方对阿霉素肾病大鼠足细胞Desmin及CD2AP蛋白的影响[J].中国中西医结合杂志,2014,34(2):203-208.WANG Z,LIU J T,SUN W S,et al.Effect of Qufeng Tongluo Recipe on expression of desmin and CD2AP proteins in adriamycin-induced nephropathy rats:an experimental research[J].Chinese Journal of Integrated Traditional and Western Medicine,2014,34(2):203-208.
[25] WANG Z,LIU J T,SUN W S.Effects of asiaticoside on levels of podocyte cytoskeletal proteins and renal slit diaphragm proteins in adriamycin-induced rat nephropathy[J].Life Sci,2013,93(8):352-358.
[26] SUN D,ZHAO X,MENG L.Relationship between urinary podocytes and kidney diseases[J].Ren Fail,2012,34(3):403-407.
[27] WU X M,GAO Y B,XU L P,et al.Exosomes from high glucose-treated glomerular endothelial cells trigger the epithelial-mesenchymal transition and dysfunction of podocytes[J].Sci Rep,2017,7(1):9371.
[2] OLATUNJI O J,CHEN H X,ZHOU Y F.Lycium chinense leaves extract ameliorates diabetic nephropathy by suppressing hyperglycemia mediated renal oxidative stress and inflammation[J].Biomed Pharmacother,2018,102:1145-1151.
[3] ZHANG X L,HE H,LIANG D,et al.Protective effects of berberine on renal injury in streptozotocin (STZ)-induced diabetic mice[J].Int J Mol Sci,2016,17(8):E1327.
[4] SAULNIER-BLACHE J S,FEIGERLOVA E,HALIMI J M,et al.Urinary lysophopholipids are increased in diabetic patients with nephropathy[J].J Diabetes Complications,2017,31(7):1103-1108.
[5] ZHAO L,WANG X L,SUN L N,et al.Critical role of serum response factor in podocyte epithelial-mesenchymal transition of diabetic nephropathy[J].Diab Vasc Dis Res,2016,13(1):81-92.
[6] CHEN X W,DU X Y,WANG Y X,et al.Irbesartan ameliorates diabetic nephropathy by suppressing the RANKL-RANK-NF-κB pathway in type 2 diabetic db/db mice[J].Mediators Inflamm,2016,2016:1405924.
[7] LING L,CHEN L B,ZHANG C N,et al.High glucose induces podocyte epithelial-to-mesenchymal transition by demethylation-mediated enhancement of MMP9 expression[J].Mol Med Rep,2018,17(4):5642-5651.
[8] YING Q D,WU G Z.Molecular mechanisms involved in podocyte EMT and concomitant diabetic kidney diseases:an update[J].Ren Fail,2017,39(1):474-483.
[9] LIU Z H.Nephrology in China[J].Nat Rev Nephrol,2013,9(9):523-528.
[10] LI B Q,CHEN J,WU T X,et al.Fast determination of four active compounds in Sanqi Panax Notoginseng Injection samples by high-performance liquid chromatography with a chemometric method[J].J Sep Sci,2015,38(9):1449-1457.
[11] 王巧凡.银可络联合三七、黄芪颗粒治疗糖尿病肾病疗效观察[J].中国中医急症,2007,16(3):294,296.WANG Q F.Efficacy of Yinkeluo combined with panax notoginseng and Huangqi granules in the treatment of diabetic nephropathy[J].Journal of Emergency in Traditional Chinese Medicine,2007,16(3):294,296.
[12] CENTER FOR DRUG EVALUATION AND RESEARCH (CDER).Estimating the maximum safe starting dose in initial clinical for therapeutics in adult healthy volunteers[S/OL].[2019-01-20] https://www.fda.gov/regulatory-information/search-fda-guidance-documents/estimating-maximum-safe-starting-dose-initial-clinical-trials-therapeutics-adult-healthy-volunteers.
[13] GUI D K,WEI L,JIAN G H,et al.Notoginsenoside R1 ameliorates podocyte adhesion under diabetic condition through α3β1 integrin upregulation in vitro and in vivo[J].Cell Physiol Biochem,2014,34(6):1849-1862.
[14] 肖磊,王华军,郑小飞,等.糖尿病对骨科手术的不利影响[J].暨南大学学报(自然科学与医学版),2018,39(5):426-434.XIAO L,WANG H J,ZHENG X F,et al.Overview of the pathology and adverse impacts of diabetes on orthopedic surgery[J].Journal of Jinan University(Natural Science & Medicine Edition),2018,39(5):426-434.
[15] 朱炳铭,吴勇,余权,等.血清miR-21联合尿微量白蛋白、尿微量白蛋白与尿肌酐比值诊断糖尿病肾病的价值[J].暨南大学学报(自然科学与医学版),2016,37(4):318-323.ZHU B M,WU Y,YU Q,et al.Clinical values of serum miR-21(SmiR-21) combined with urinary micro-albumin (UmAlb) and UmAlb /urine creatinine (UmAlb/Ucr) for diagnosis of diabetic nephropathy[J].Journal of Jinan University(Natural Science & Medicine Edition),2016,37(4):318-323.
[16] STINGHEN A E,MASSY Z A,VLASSARA H,et al.Uremic toxicity of advanced glycation end products in CKD[J].J Am Soc Nephrol,2016,27(2):354-370.
[17] 占永力,岳玉和,周静媛,等.中药三七注射液对显性糖尿病肾病并高凝状态的影响[J].中国中西医结合肾病杂志,2002,3(1):23-25.ZHAN Y L,YUE Y H,ZHOU J Y,et al.Effect of Xue Shuan Tong injection on overt diabetic nephropathy with hypercoagulability[J].Chinese Journal of Integrated Traditional and Western Nephrology,2002,3(1):23-25.
[18] 郑丽阳,薛瑞,吕亭亭,等.三七皂苷Fc对2型糖尿病db/db小鼠的肾脏保护作用研究[J].中华中医药学刊,2017,35(3):609-611.ZHENG L Y,XUE R,Lü T T,et al.Study on renal protective effect of notoginsenoside fc on db/db mice[J].Chinese Archives of Traditional Chinese Medicine,2017,35(3):609-611.
[19] HUANG G D,LV J,LI T Y,et al.Notoginsenoside R1 ameliorates podocyte injury in rats with diabetic nephropathy by activating the PI3K/Akt signaling pathway[J].Int J Mol Med,2016,38(4):1179-1189.
[20] 王洪敏,钟志勇,吴庆洲,等.大鼠糖尿病模型早期病理观察指标的筛选[J].广东药学院学报,2011,27(1):86-90.WANG H M,ZHONG Z Y,WU Q Z,et al.Selection of pathogenic indicators in the early stage of diabetes rats[J].Journal of Guangdong Pharmaceutical College,2011,27(1):86-90.
[21] MUNDEL P.Podocytes and the quest for precision medicines for kidney diseases[J].Pflugers Arch,2017,469(7/8):1029-1037.
[22] KANG Y S,LI Y J,DAI C S,et al.Inhibition of integrin-linked kinase blocks podocyte epithelial-mesenchymal transition and ameliorates proteinuria[J].Kidney Int,2010,78(4):363-373.
[23] DAI H R,LIU Q Q,LIU B L.Research progress on mechanism of podocyte depletion in diabetic nephropathy[J].J Diabetes Res,2017,2017:2615286.
[24] 王竹,刘俊田,孙万森,等.祛风通络方对阿霉素肾病大鼠足细胞Desmin及CD2AP蛋白的影响[J].中国中西医结合杂志,2014,34(2):203-208.WANG Z,LIU J T,SUN W S,et al.Effect of Qufeng Tongluo Recipe on expression of desmin and CD2AP proteins in adriamycin-induced nephropathy rats:an experimental research[J].Chinese Journal of Integrated Traditional and Western Medicine,2014,34(2):203-208.
[25] WANG Z,LIU J T,SUN W S.Effects of asiaticoside on levels of podocyte cytoskeletal proteins and renal slit diaphragm proteins in adriamycin-induced rat nephropathy[J].Life Sci,2013,93(8):352-358.
[26] SUN D,ZHAO X,MENG L.Relationship between urinary podocytes and kidney diseases[J].Ren Fail,2012,34(3):403-407.
[27] WU X M,GAO Y B,XU L P,et al.Exosomes from high glucose-treated glomerular endothelial cells trigger the epithelial-mesenchymal transition and dysfunction of podocytes[J].Sci Rep,2017,7(1):9371.