加味补阳还五汤对七氟烷麻醉处理后早期糖尿病肾病大鼠肾足细胞影响实验研究
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摘要
目的:探讨加味补阳还五汤(MDIY)对七氟烷麻醉处理后的早期糖尿病肾病(DN)大鼠肾足细胞的影响。方法:将早期DN大鼠随机分为4组(每组n=16):MDIY组(A组),罗格列酮组(B组),MDIY+罗格列酮组(C组)和模型对照组(D组),另设正常大鼠(E组)为空白对照组。连续治疗7周,每天收集大鼠死亡数和葡萄糖(GLU)、肌酐(CR)、尿素氮(BUN)和尿微量白蛋白(UAL)。七氟烷麻醉处理后采集标本,通过苏木精和伊红染色(H&E)收集肾脏和肾病理变化检查。应用实RT-PCR和westernblotting检测评估肾足突细胞各m-RNA的表达水平,包括nephrin、podocin、α-SMA和desmin的表达。结果:在治疗周期中,A组、B组和C组的GLU和UAL水平均低于D组(P<0.01),且7周后3组的GLU、CR和BUN水平也都远低于D组(P<0.05)。其中,C组的上述生化指标远低于A组和B组(P<0.05)。相比于正常SD大鼠,通过H&E染色,观察各组的肾小球疾病、慢性肾小管间质损伤和肾小球足细胞损伤的发病率。然而,只有C组显示更能优化改善上述损伤的发病率。在A组、B组和C组与D组比较,α-SMA和desmin的表达减少,而nephrin和podocin的表达明显增加(P<0.05);C组不仅nephrin和podocin mRNA表达(P<0.05),而且在蛋白表达方面均观察有协同效应。结论:MDIY对早期DN引起的肾功能损害是一种保护作用,可以大大提高与罗格列酮相结合的治疗效果,其机制是通过增加nephrin、podocin的表达和减少α-SMA、desmin的表达,从而推测MDIY具有显著的协同治疗DN作用。七氟烷预处理能改善糖尿病肾病足细胞结构和功能,对围术期外科快速康复有利。
Objective:To expore the effects and mechanism of modified Buyang Huanwu Decoction reducing the risk of kidney damage caused by the early of the diabetic nephropathy(DN)with sevoflurane preconditioning. Methods:Then DN rats were randomly divided into four groups(n=16),MDIY(group A),rosiglitazone(group B),MDIY+ rosiglitazone(group C)and normal saline(group D). During the treating cycle,the glucose(GLU)and urine albumin(UAL)were detected continuously. After the treatment of 7 weeks,the rats were sacrificed and the level of GLU,serum creatinine(SCR)and blood urea nitrogen(BUN)were detected in the serum. At the same time,the kidneys were collected and the renal pathological changes were inspected by hematoxylin and eosin(H&E)staining. The expression levels of proteinuria including nephrin,podocin,α-smooth muscle actin(α-SMA)and desmin were evaluated by Real-time PCR and Western blotting. Results:During the treating cycle,GLU and UAL in the A,B and C groups were lower than group D(P<0.01),after 7 weeks the level of GLU,CR and BUN levels in group A,B and C were much lower than group D(P<0.05). Compared with group A and group B,biochemical indexes above mentioned were much lower in group C(P<0.05). Incidences of glomerular disorders,chronic tubulointerstitial injure and glomerular podocyte damage were observed in group A,B,C and D when compared to normal SD rats by H&E staining. However,group C showed optimal improvement of glomerular disorders,chronic tubulointerstitial injure and glomerular podocyte damage. The expression of α-SMA and desmin decreased,while the expression of nephrin and podocin increased markedly in group A,B and C comparing with group D(P<0.05),which were detected by Real-time PCR and Western blotting. The synergistic effect was observed not only in the mRNA expression of nephrin and podocin in group C(P<0.05),but also in the protein expression. Conclusion:MDIY was proven to be a protective effect in kidney damage caused by DN and could considerably ameliorate the therapeutic efficacy combined with rosiglitazone,which developed a treating role through increasing the expression of nephrin and podocin and decreasing the expression of α-SMA and desmin. And sevoflurane preconditioning can improve the structure and function of diabetic nephropathy,and it is beneficial to the rapid rehabilitation of perioperative period.
Objective:To expore the effects and mechanism of modified Buyang Huanwu Decoction reducing the risk of kidney damage caused by the early of the diabetic nephropathy(DN)with sevoflurane preconditioning. Methods:Then DN rats were randomly divided into four groups(n=16),MDIY(group A),rosiglitazone(group B),MDIY+ rosiglitazone(group C)and normal saline(group D). During the treating cycle,the glucose(GLU)and urine albumin(UAL)were detected continuously. After the treatment of 7 weeks,the rats were sacrificed and the level of GLU,serum creatinine(SCR)and blood urea nitrogen(BUN)were detected in the serum. At the same time,the kidneys were collected and the renal pathological changes were inspected by hematoxylin and eosin(H&E)staining. The expression levels of proteinuria including nephrin,podocin,α-smooth muscle actin(α-SMA)and desmin were evaluated by Real-time PCR and Western blotting. Results:During the treating cycle,GLU and UAL in the A,B and C groups were lower than group D(P<0.01),after 7 weeks the level of GLU,CR and BUN levels in group A,B and C were much lower than group D(P<0.05). Compared with group A and group B,biochemical indexes above mentioned were much lower in group C(P<0.05). Incidences of glomerular disorders,chronic tubulointerstitial injure and glomerular podocyte damage were observed in group A,B,C and D when compared to normal SD rats by H&E staining. However,group C showed optimal improvement of glomerular disorders,chronic tubulointerstitial injure and glomerular podocyte damage. The expression of α-SMA and desmin decreased,while the expression of nephrin and podocin increased markedly in group A,B and C comparing with group D(P<0.05),which were detected by Real-time PCR and Western blotting. The synergistic effect was observed not only in the mRNA expression of nephrin and podocin in group C(P<0.05),but also in the protein expression. Conclusion:MDIY was proven to be a protective effect in kidney damage caused by DN and could considerably ameliorate the therapeutic efficacy combined with rosiglitazone,which developed a treating role through increasing the expression of nephrin and podocin and decreasing the expression of α-SMA and desmin. And sevoflurane preconditioning can improve the structure and function of diabetic nephropathy,and it is beneficial to the rapid rehabilitation of perioperative period.
引文
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[4]Yingying Sun,Yuanhai Li,Lei Liu,et al. Identification of miRNAs Involved in the Protective Effect of Sevoflurane Preconditioning Against Hypoxic Injury in PC12 Cells[J].Cellular and Molecular Neurobiology,2015,35(8):1117-1125.
[5]Zhang Z,Wu F,Zheng F,et al. Adenovirus-mediated decorin gene transfection has therapeutic effects in a streptozocin-induced diabetic rat model[J]. Nephron Experimental Nephrology,2010,116(1):11-21.
[6]Srinivasan M,Blackburn C,MohamedM,et al. Literature-based discovery of salivary biomarkers for type 2 diabetes mellitus[J].Biomarker Insights,2015,10(1):39-45.
[7]Liu Willow JH. What has been overlooked on study of Chinese materia medica in the West[J]. Chinese Journal of Integrative Medicine,2015,21(7):483-492.
[8]Lim Andy KH.Diabetic nephropathy-complications and treatment[J].InternationalJournalofNephrologyand Renovascular Disease,2014,10(7):361-381.
[9]EffersonJA,ShanklandSJ,PichlerRH.Proteinuriain diabetic kidney disease:a mechanistic view point[J]. Kidney International,2008,74(1):22-36.
[10]Zhang YK,Han XY,Che ZY.Effects of buyang huanwu tang combined with bone marrow mesenchymal stem cell transplantation on the expression of VEGF and Ki-67 in the brain tissue of the cerebral ischemia-reperfusion model rat[J].Journal of traditional Chinese medicine,2010,30(4):278-282.
[11]Zhou Y C,LiuB,LiYJ,et al. Effects of buyang huanwu decoction on ventricular remodeling and differential protein profile in a rat model of myocardial infarction[J]. Evidencebased Complementary and Alternative Medicine,2012,1155(2012):385247-385258.
[12]Wu L,Zhang W,LiH,et al.Inhibition of aortic intimal hyperplasia and cell cycle protein and extracellular matrix protein expressions by BuYang HuanWu Decoction[J]. Journal of Ethnopharmacology,2009,125(3):423-435.
[13]Cai XL,Chen Y L,Zhao JJ,et al.Efficacy and Safety of Avandamet or Uptitrated Metformin Treatment in Patients with Type 2 Diabetes Inadequately Controlled with Metformin Alone:A Multicenter,Randomized,Controlled Trial[J]. Chinese Medical Journal,2015,128(10):1279-1287.
[14]Chodavarapu H,Grobe N,Somineni HK,et al.Rosiglitazone treatment of type 2 diabetic db/db mice attenuates urinary albumin and angiotensin converting enzyme 2 excretion[J].PloS One,2013,8(4):e62833.
[15]Xing N,Wei X,Chang YZ,et al. Effects of low-flow sevoflurane anesthesia on renal function in low birth weight infants[J]. BMC Anesthesiology,2015,15(15):2151-2155.
[16]Lu HJ,Tzeng TF,Liou SS,et al. Ruscogenin ameliorates diabetic nephropathy by its anti-inflammatory and antifibrotic effects in streptozotocin-induced diabetic rat[J].BMC Complementary and Alternative Medicine,2014,14(3):110-122.
[17] Zhang X,Song Z,Guo Y,et al.The novel role of TRPC6 in vitamin D ameliorating podocyte injury in STZ-induced diabetic rats[J]. Molecular and Cellular Biochemistry,2015,399(2):155-165.
[18] Jourdan T,Szanda G,Rosenberg AZ,et al. Overactive cannabinoid 1 receptor in podocytes drives type 2 diabetic nephropathy[J]. Proceedings of the National Academy of Sciences of the United States of America,2014,111(50):e5420-e5428.
[19] Liu Y,Long L,Yuan F,et al. High glucose-induced Galectin-1in human podocytes implicates the involvement of Galectin-1 in diabetic nephropathy[J].Cell Biology International,2015,39(2):217-223.
[20] Peng T,Wang J,Zhen J,et al.Effect of benazepril on the transdifferentiation of renal tubular epithelial cells from diabetic rats[J]. Biomedical Reports,2014,2(4):490-494.
[21] Niu H,Nie L,Liu M,et al. Benazepril affects integrin-linked kinase and smooth muscle alpha-actin expression in diabetic rat glomerulus and cultured mesangial cells[J]. BMC Nephrology,2014,15(2):135-145.
[22] Ouyang B,Sun X,Han D,et al. Human urine-derived stem cells alone or genetically-modified with FGF2 Improve type 2 diabetic erectile dysfunction in a rat model[J]. PlOS One,2014,9(3):e92825.
[23] Hathaway CK,Gasim A M,Grant R,et al. Low TGF beta1expression prevents and high expression exacerbates diabetic nephropathy in mice[J]. Proceedings of the National Academy of Sciences of the United States of America,2015,112(18):5815-5820.
[24] Liu W,Qiao F,Liu H,et al. Low molecular weight heparin improves proteinuria in rats with L-NAME induced preeclampsia by decreasing the expression of nephrin,but not podocin[J].Hypertension In Pregnancy,2015,34(1):24-35.
[25] Sanai T,Sobka T,Johnson T,et al. Expression of cytoskeletal proteins during the course of experimental diabetic nephropathy[J]. Diabetologia,2000,43(1):91-100.
[26] Xing L,Liu Q,Fu S,et al. PTEN inhibits high glucose-induced phenotypic transition in podocytes[J]. Journal of Cellular Biochemistry,2015,116(8):1776-1784.
[2]Yoshiteru Mori,Takayuki Kitamura,Gaku Kawamura,et al.Effects of preoperative and intraoperative glucose administration on glucose use and fat catabolism during laparotomy under sevoflurane anesthesia in fasted rats[J].The Journal of Physiological Sciences,2015,65(6):523-530.
[3]Kong X1,Su X,Zhu J,et al. Neuroprotective effect of buyang huanwu decoction on rat ischemic/reperfusion brain damage by promoting migration of neural precursor cells[J]. Rejuvenation Res, 2014,17(3):264-275.
[4]Yingying Sun,Yuanhai Li,Lei Liu,et al. Identification of miRNAs Involved in the Protective Effect of Sevoflurane Preconditioning Against Hypoxic Injury in PC12 Cells[J].Cellular and Molecular Neurobiology,2015,35(8):1117-1125.
[5]Zhang Z,Wu F,Zheng F,et al. Adenovirus-mediated decorin gene transfection has therapeutic effects in a streptozocin-induced diabetic rat model[J]. Nephron Experimental Nephrology,2010,116(1):11-21.
[6]Srinivasan M,Blackburn C,MohamedM,et al. Literature-based discovery of salivary biomarkers for type 2 diabetes mellitus[J].Biomarker Insights,2015,10(1):39-45.
[7]Liu Willow JH. What has been overlooked on study of Chinese materia medica in the West[J]. Chinese Journal of Integrative Medicine,2015,21(7):483-492.
[8]Lim Andy KH.Diabetic nephropathy-complications and treatment[J].InternationalJournalofNephrologyand Renovascular Disease,2014,10(7):361-381.
[9]EffersonJA,ShanklandSJ,PichlerRH.Proteinuriain diabetic kidney disease:a mechanistic view point[J]. Kidney International,2008,74(1):22-36.
[10]Zhang YK,Han XY,Che ZY.Effects of buyang huanwu tang combined with bone marrow mesenchymal stem cell transplantation on the expression of VEGF and Ki-67 in the brain tissue of the cerebral ischemia-reperfusion model rat[J].Journal of traditional Chinese medicine,2010,30(4):278-282.
[11]Zhou Y C,LiuB,LiYJ,et al. Effects of buyang huanwu decoction on ventricular remodeling and differential protein profile in a rat model of myocardial infarction[J]. Evidencebased Complementary and Alternative Medicine,2012,1155(2012):385247-385258.
[12]Wu L,Zhang W,LiH,et al.Inhibition of aortic intimal hyperplasia and cell cycle protein and extracellular matrix protein expressions by BuYang HuanWu Decoction[J]. Journal of Ethnopharmacology,2009,125(3):423-435.
[13]Cai XL,Chen Y L,Zhao JJ,et al.Efficacy and Safety of Avandamet or Uptitrated Metformin Treatment in Patients with Type 2 Diabetes Inadequately Controlled with Metformin Alone:A Multicenter,Randomized,Controlled Trial[J]. Chinese Medical Journal,2015,128(10):1279-1287.
[14]Chodavarapu H,Grobe N,Somineni HK,et al.Rosiglitazone treatment of type 2 diabetic db/db mice attenuates urinary albumin and angiotensin converting enzyme 2 excretion[J].PloS One,2013,8(4):e62833.
[15]Xing N,Wei X,Chang YZ,et al. Effects of low-flow sevoflurane anesthesia on renal function in low birth weight infants[J]. BMC Anesthesiology,2015,15(15):2151-2155.
[16]Lu HJ,Tzeng TF,Liou SS,et al. Ruscogenin ameliorates diabetic nephropathy by its anti-inflammatory and antifibrotic effects in streptozotocin-induced diabetic rat[J].BMC Complementary and Alternative Medicine,2014,14(3):110-122.
[17] Zhang X,Song Z,Guo Y,et al.The novel role of TRPC6 in vitamin D ameliorating podocyte injury in STZ-induced diabetic rats[J]. Molecular and Cellular Biochemistry,2015,399(2):155-165.
[18] Jourdan T,Szanda G,Rosenberg AZ,et al. Overactive cannabinoid 1 receptor in podocytes drives type 2 diabetic nephropathy[J]. Proceedings of the National Academy of Sciences of the United States of America,2014,111(50):e5420-e5428.
[19] Liu Y,Long L,Yuan F,et al. High glucose-induced Galectin-1in human podocytes implicates the involvement of Galectin-1 in diabetic nephropathy[J].Cell Biology International,2015,39(2):217-223.
[20] Peng T,Wang J,Zhen J,et al.Effect of benazepril on the transdifferentiation of renal tubular epithelial cells from diabetic rats[J]. Biomedical Reports,2014,2(4):490-494.
[21] Niu H,Nie L,Liu M,et al. Benazepril affects integrin-linked kinase and smooth muscle alpha-actin expression in diabetic rat glomerulus and cultured mesangial cells[J]. BMC Nephrology,2014,15(2):135-145.
[22] Ouyang B,Sun X,Han D,et al. Human urine-derived stem cells alone or genetically-modified with FGF2 Improve type 2 diabetic erectile dysfunction in a rat model[J]. PlOS One,2014,9(3):e92825.
[23] Hathaway CK,Gasim A M,Grant R,et al. Low TGF beta1expression prevents and high expression exacerbates diabetic nephropathy in mice[J]. Proceedings of the National Academy of Sciences of the United States of America,2015,112(18):5815-5820.
[24] Liu W,Qiao F,Liu H,et al. Low molecular weight heparin improves proteinuria in rats with L-NAME induced preeclampsia by decreasing the expression of nephrin,but not podocin[J].Hypertension In Pregnancy,2015,34(1):24-35.
[25] Sanai T,Sobka T,Johnson T,et al. Expression of cytoskeletal proteins during the course of experimental diabetic nephropathy[J]. Diabetologia,2000,43(1):91-100.
[26] Xing L,Liu Q,Fu S,et al. PTEN inhibits high glucose-induced phenotypic transition in podocytes[J]. Journal of Cellular Biochemistry,2015,116(8):1776-1784.