消瘀泄浊饮对造影剂肾病大鼠保护作用的实验研究
|
摘要
目的探讨消瘀泄浊饮对造影剂肾病(CIN)大鼠是否具有保护作用及其相关机制。方法将24只雄性SD大鼠随机分为4组:正常组(A组)、CIN组(B组)、消瘀泄浊饮低剂量组(C组)和消瘀泄浊饮高剂量组(D组)。造模后48 h处死大鼠,测定各组大鼠血清肌酐、尿素氮水平,HE和PAS染色观察各组肾脏组织病理变化,Western Blot检测各组Bax、Bcl-2、Caspase-3、P-Akt/Akt和P-mTOR/mTOR蛋白表达量。结果造模48 h后,与A组相比,B组大鼠血清肌酐和尿素氮水平均明显升高(P<0.05);肾小管上皮细胞出现空泡样变性、管腔变大及肾间质水肿,肾小球组织未发现明显改变;Bax/Bcl-2和Caspase-3蛋白表达上调(P<0.05),且P-Akt/Akt和P-mTOR/mTOR蛋白表达同时上调(P<0.05)。与B组相比,C、D组血清肌酐和尿素氮水平明显下降(P<0.05);肾小管上皮空泡样变性减少;Bax/Bcl-2和Caspase-3蛋白表达下调(P<0.05),且P-Akt/Akt和P-m TOR/mTOR蛋白表达同时下调(P<0.05)。结论消瘀泄浊饮对造影剂肾病大鼠具有预防作用,其机制可能是通过抑制PI3K/Akt/mTOR信号通路过度激活从而抑制肾小管上皮细胞凋亡。
Objective To investigate whether Xiaoyu Xiezhuo decoction has protective effect on contrast-induced nephropathy(CIN) rats and its related mechanisms.Methods 24 male Sprague-Dawley rats were randomly divided into4 groups:normal group(group A),CIN group(group B),Xiaoyu Xiezhuo low-dose group(group C) and Xiaoyu Xiezhuo high-dose group(Group D).The rats were sacrificed at 48 h after model establishment.Serum creatinine and urea nitrogen levels of rats in each group were measured.The pathological changes of renal tissues were observed by HE and PAS.The Bax,Bcl-2,Caspase-3 and P-Akt/Akt and P-mTOR/mTOR protein expression levels were detected by Western Blot.Results After 48 hours of modeling,serum creatinine and urea nitrogen levels in group B were significantly higher than those of group A(P<0.05).Compared with those in group A,vacuolar degeneration,lumen enlargement and renal interstitial in group B occurred in renal tubular epithelial cells,and there was no obvious change in edema and glomerular tissue.Compared with those in group A,Bax/Bcl-2 and Caspase-3 protein expression was upregulated(P<0.05),and P-Akt/Akt and P-mTOR/mTOR protein expression were up-regulated simultaneously in group B(P<0.05).Compared with those of group B,serum creatinine and urea nitrogen levels in group C and D were significantly decreased(P<0.05);vacuolar degeneration in renal tubular epithelium was decreased;Bax/Bcl-2 and Caspase-3 protein expression were down-regulated(P<0.05);and P-Akt/Akt and P-mTOR/mTOR protein expression were simultaneously down-regulated(P<0.05).Conclusion Xiaoyu Xiezhuo decoction has a preventive effect on rats with contrastinduced nephropathy.The mechanism may be to inhibit the apoptosis of renal tubular epithelial cells by inhibiting the excessive activation of PI3 K/Akt/mTOR signaling pathway.
Objective To investigate whether Xiaoyu Xiezhuo decoction has protective effect on contrast-induced nephropathy(CIN) rats and its related mechanisms.Methods 24 male Sprague-Dawley rats were randomly divided into4 groups:normal group(group A),CIN group(group B),Xiaoyu Xiezhuo low-dose group(group C) and Xiaoyu Xiezhuo high-dose group(Group D).The rats were sacrificed at 48 h after model establishment.Serum creatinine and urea nitrogen levels of rats in each group were measured.The pathological changes of renal tissues were observed by HE and PAS.The Bax,Bcl-2,Caspase-3 and P-Akt/Akt and P-mTOR/mTOR protein expression levels were detected by Western Blot.Results After 48 hours of modeling,serum creatinine and urea nitrogen levels in group B were significantly higher than those of group A(P<0.05).Compared with those in group A,vacuolar degeneration,lumen enlargement and renal interstitial in group B occurred in renal tubular epithelial cells,and there was no obvious change in edema and glomerular tissue.Compared with those in group A,Bax/Bcl-2 and Caspase-3 protein expression was upregulated(P<0.05),and P-Akt/Akt and P-mTOR/mTOR protein expression were up-regulated simultaneously in group B(P<0.05).Compared with those of group B,serum creatinine and urea nitrogen levels in group C and D were significantly decreased(P<0.05);vacuolar degeneration in renal tubular epithelium was decreased;Bax/Bcl-2 and Caspase-3 protein expression were down-regulated(P<0.05);and P-Akt/Akt and P-mTOR/mTOR protein expression were simultaneously down-regulated(P<0.05).Conclusion Xiaoyu Xiezhuo decoction has a preventive effect on rats with contrastinduced nephropathy.The mechanism may be to inhibit the apoptosis of renal tubular epithelial cells by inhibiting the excessive activation of PI3 K/Akt/mTOR signaling pathway.
引文
[1] Finn WF. The clinical and renal consequences of contrastinduced nephropathy[J]. Nephrol Dial Transplant,2006,21(6):2-10.
[2] Toprak O,Cirit M. Risk factors and therapy strategies for contrast-induced nephropathy[J]. Ren Fail,2006,28(5):365-381.
[3] Humbert A,Kissling S,Teta D. Contrast-induced nephropathy[J]. Indian Heart Journal, 2014,21(2):261-280.
[4]鲁科达,邵明祥,马红珍,等.消瘀泄浊饮治疗慢性肾脏病气虚夹瘀证临床疗效观察[J].浙江中医药大学学报,2014,38(6):718-721.
[5] Wang F,Zhang G,Zhou Y,et al. Magnolin protects against contrast-induced nephropathy in rats via antioxidation and antiapoptosis[J]. Oxidative Medicine&Cellular Longevity,2016,2014:203458.
[6] Liang R,Zhao Q,Jian G,et al. Tanshinone IIA Attenuates Contrast-Induced Nephropathy via Nrf2 Activation in Rats[J]. Cellular Physiology&Biochemistry International Journal of Experimental Cellular Physiology Biochemistry&Pharmacology,2018,46(6):2616.
[7] He X,Li L,Tan H,et al. Atorvastatin attenuates contrast-induced nephropathy by modulating inflammatory responses through the regulation of JNK/p38/Hsp27 expression[J]. Journal of Pharmacological Sciences,2016,131(1):18-27.
[8] Stacul F,Molen AJVD,Reimer P,et al. Contrast induced nephropathy:updated ESUR Contrast Media Safety Committee guidelines[J]. European Radiology,2012,21(12):2527-2541.
[9] Maioli M,Toso A,Leoncini M,et al. Persistent renal damage after contrast-induced acute kidney injury:Incidence,evolution,risk factors,and prognosis[J]. Circulation,2012,125(25):3099.
[10] Sendeski M,Patzak A,Pallone TL,et al. Iodixanol,constriction of medullary descending vasa recta,and risk for contrast medium-induced nephropathy[J]. Radiology,2009,251(3):697-704.
[11] Andreucci M,Faga T,Russo D,et al. Differential activation of signaling pathways by low-osmolar and iso-osmolar radiocontrast agents in human renal tubular cells[J].Journal of Cellular Biochemistry,2013,115(2):281-289.
[12] Quintavalle C,Brenca M,Micco FD,et al. In vivo and in vitro assessment of pathways involved in contrast mediainduced renal cells apoptosis[J]. Cell Death&Disease,2011,2(5):e155.
[13]鲁科达,叶黎青,陈红波,等.李学铭运用消瘀泄浊饮治疗肾病的经验[J].中华中医药学刊,2011,(8):1745-1746.
[14] Yokomaku Y,Sugimoto T,Kume S,et al. Asialoerythropoietin prevents contrast-induced nephropathy[J]. Journal of the American Society of Nephrology Jasn,2008,19(2):321.
[15] Meier P,Finch A,Evan G. Apoptosis in development[J].Nature,2000,407(6805):796-801.
[16] Zhao H,Yenari MA,Cheng D,et al. Bcl-2 overexpression protects against neuron loss within the ischemic margin following experimental stroke and inhibits cytochrome c translocation and caspase-3 activity[J]. Journal of Neurochemistry,2003,85(4):1026-1036.
[17] Hubbard PA,Moody CL,Murali R. Allosteric modulation of Ras and the PI3K/AKT/mTOR pathway:emerging therapeutic opportunities[J]. Frontiers in Physiology,2014,5:478.
[18] Dormond O,Madsen JC,Briscoe DM. The effects of m TOR-Akt interactions on anti-apoptotic signaling in vascular endothelial cells[J]. Journal of Biological Chemistry, 2007,282(32):23679.
[19] Zhao G,Han Y,Hu Y,et al. Targeting HO-1 by Epigallocatechin-3-Gallate Reduces Contrast-Induced Renal Injury via Anti-Oxidative Stress and Anti-Inflammation Pathways[J]. Plos One,2016,11(2):e0149032.
[20] Michael A,Faga T,Pisani A,et al. Molecular Mechanisms of Renal Cellular Nephrotoxicity due to Radiocontrast Media[J]. Biomed Research International,2014,(6):249810.
[21] He X,Yang J,Li L,et al. Atorvastatin protects against contrast-induced nephropathy via anti-apoptosis by the upregulation of Hsp27in vivoandin vitro[J]. Molecular Medicine Reports,2017,15(4):1963-1972.
[2] Toprak O,Cirit M. Risk factors and therapy strategies for contrast-induced nephropathy[J]. Ren Fail,2006,28(5):365-381.
[3] Humbert A,Kissling S,Teta D. Contrast-induced nephropathy[J]. Indian Heart Journal, 2014,21(2):261-280.
[4]鲁科达,邵明祥,马红珍,等.消瘀泄浊饮治疗慢性肾脏病气虚夹瘀证临床疗效观察[J].浙江中医药大学学报,2014,38(6):718-721.
[5] Wang F,Zhang G,Zhou Y,et al. Magnolin protects against contrast-induced nephropathy in rats via antioxidation and antiapoptosis[J]. Oxidative Medicine&Cellular Longevity,2016,2014:203458.
[6] Liang R,Zhao Q,Jian G,et al. Tanshinone IIA Attenuates Contrast-Induced Nephropathy via Nrf2 Activation in Rats[J]. Cellular Physiology&Biochemistry International Journal of Experimental Cellular Physiology Biochemistry&Pharmacology,2018,46(6):2616.
[7] He X,Li L,Tan H,et al. Atorvastatin attenuates contrast-induced nephropathy by modulating inflammatory responses through the regulation of JNK/p38/Hsp27 expression[J]. Journal of Pharmacological Sciences,2016,131(1):18-27.
[8] Stacul F,Molen AJVD,Reimer P,et al. Contrast induced nephropathy:updated ESUR Contrast Media Safety Committee guidelines[J]. European Radiology,2012,21(12):2527-2541.
[9] Maioli M,Toso A,Leoncini M,et al. Persistent renal damage after contrast-induced acute kidney injury:Incidence,evolution,risk factors,and prognosis[J]. Circulation,2012,125(25):3099.
[10] Sendeski M,Patzak A,Pallone TL,et al. Iodixanol,constriction of medullary descending vasa recta,and risk for contrast medium-induced nephropathy[J]. Radiology,2009,251(3):697-704.
[11] Andreucci M,Faga T,Russo D,et al. Differential activation of signaling pathways by low-osmolar and iso-osmolar radiocontrast agents in human renal tubular cells[J].Journal of Cellular Biochemistry,2013,115(2):281-289.
[12] Quintavalle C,Brenca M,Micco FD,et al. In vivo and in vitro assessment of pathways involved in contrast mediainduced renal cells apoptosis[J]. Cell Death&Disease,2011,2(5):e155.
[13]鲁科达,叶黎青,陈红波,等.李学铭运用消瘀泄浊饮治疗肾病的经验[J].中华中医药学刊,2011,(8):1745-1746.
[14] Yokomaku Y,Sugimoto T,Kume S,et al. Asialoerythropoietin prevents contrast-induced nephropathy[J]. Journal of the American Society of Nephrology Jasn,2008,19(2):321.
[15] Meier P,Finch A,Evan G. Apoptosis in development[J].Nature,2000,407(6805):796-801.
[16] Zhao H,Yenari MA,Cheng D,et al. Bcl-2 overexpression protects against neuron loss within the ischemic margin following experimental stroke and inhibits cytochrome c translocation and caspase-3 activity[J]. Journal of Neurochemistry,2003,85(4):1026-1036.
[17] Hubbard PA,Moody CL,Murali R. Allosteric modulation of Ras and the PI3K/AKT/mTOR pathway:emerging therapeutic opportunities[J]. Frontiers in Physiology,2014,5:478.
[18] Dormond O,Madsen JC,Briscoe DM. The effects of m TOR-Akt interactions on anti-apoptotic signaling in vascular endothelial cells[J]. Journal of Biological Chemistry, 2007,282(32):23679.
[19] Zhao G,Han Y,Hu Y,et al. Targeting HO-1 by Epigallocatechin-3-Gallate Reduces Contrast-Induced Renal Injury via Anti-Oxidative Stress and Anti-Inflammation Pathways[J]. Plos One,2016,11(2):e0149032.
[20] Michael A,Faga T,Pisani A,et al. Molecular Mechanisms of Renal Cellular Nephrotoxicity due to Radiocontrast Media[J]. Biomed Research International,2014,(6):249810.
[21] He X,Yang J,Li L,et al. Atorvastatin protects against contrast-induced nephropathy via anti-apoptosis by the upregulation of Hsp27in vivoandin vitro[J]. Molecular Medicine Reports,2017,15(4):1963-1972.