丹参与环孢素A对移植肾缺血再灌注损伤防治作用的对比研究
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摘要
目的:探讨药物对移植肾缺血再灌注损伤(IRI)的防治作用。
方法:40只健康雄性日本大耳兔随机分为四组:生理盐水对照组、单纯灌注液组、丹参+灌注液组、环孢素A(CsA)+灌注液组。用套管针穿刺左肾动、静脉,形成局部循环,灌注后原位低温保存2小时,结扎并切除右肾后使左肾复灌。于术后6、24小时分别取血标本,24小时后取左肾标本。测定血尿素氮(BUN)、血肌酐(Cr);TUNEL法检测左肾组织的细胞凋亡;免疫组化法测定左肾组织热休克蛋白70(HSP70)及核转录因子κB(NF-κB)的表达。
结果:丹参+灌注液组与其他组比较,能明显降低BUN、Cr的含量(P<0.01),上调超氧化物歧化酶(SOD)活性及降低丙二醛(MDA)的产生(P<0.01)。TUNEL细胞凋亡显示与其他组相比,丹参+灌注液组与CsA+灌注液组均能减少细胞凋亡(P<0.01),但二者之间无显著性差异(P>0.05)。CsA+灌注液组与其他组相比能显著上调移植肾HSP70的表达,抑制NF-κB的活性(P<0.01)。
结论:IRI是由多因素、多环节共同作用的结果,丹参直接抑制IRI中氧自由基对移植肾的损伤,而CsA则是通过诱导HSP70表达上调来发挥对移植肾的保护作用。
Objective: To investigate the prevention of drug to the renal ischemia/reperfusion injury (IRI).Methods: Forty rabbits were divided into four groups at random. Clipped the kidney artery and vein with untrauma vascular clamp and intubated. After perfusion and preservation of left kidney at low temperature in situ for 2 hours, resected right kidney and then reperfused the left kidney. Got blood and assayed BUN、Cr at 6h、 24h after surgery. Assayed HSP70、 NF-κB by immunohistochemistry and apoptotic cell in left renal tissue by TUNEL.Results: Both perfused dan-shen root group and CsA group decreased BUN、 Gr、 MDA than that of other groups (P<0.01), and increased activity of SOD (PO.Ol). Contrast with other groups, both perfused dan-shen root group and CsA group decreased the cell apoptosis (P<0.01), but there was no significant difference between them (P>0.05). The expression of positive HSP70 in perfused CsA group was higher than that of other groups (P<0.01), and the activity of NF-κB was lower than that of other groups (PO.Ol).Conclusion: The dan-shen root can directly inhibit the injury of OFR to transplanted renal, and CsA can increase the expression of HSP70 to prevent the transplanted renal.
方法:40只健康雄性日本大耳兔随机分为四组:生理盐水对照组、单纯灌注液组、丹参+灌注液组、环孢素A(CsA)+灌注液组。用套管针穿刺左肾动、静脉,形成局部循环,灌注后原位低温保存2小时,结扎并切除右肾后使左肾复灌。于术后6、24小时分别取血标本,24小时后取左肾标本。测定血尿素氮(BUN)、血肌酐(Cr);TUNEL法检测左肾组织的细胞凋亡;免疫组化法测定左肾组织热休克蛋白70(HSP70)及核转录因子κB(NF-κB)的表达。
结果:丹参+灌注液组与其他组比较,能明显降低BUN、Cr的含量(P<0.01),上调超氧化物歧化酶(SOD)活性及降低丙二醛(MDA)的产生(P<0.01)。TUNEL细胞凋亡显示与其他组相比,丹参+灌注液组与CsA+灌注液组均能减少细胞凋亡(P<0.01),但二者之间无显著性差异(P>0.05)。CsA+灌注液组与其他组相比能显著上调移植肾HSP70的表达,抑制NF-κB的活性(P<0.01)。
结论:IRI是由多因素、多环节共同作用的结果,丹参直接抑制IRI中氧自由基对移植肾的损伤,而CsA则是通过诱导HSP70表达上调来发挥对移植肾的保护作用。
Objective: To investigate the prevention of drug to the renal ischemia/reperfusion injury (IRI).Methods: Forty rabbits were divided into four groups at random. Clipped the kidney artery and vein with untrauma vascular clamp and intubated. After perfusion and preservation of left kidney at low temperature in situ for 2 hours, resected right kidney and then reperfused the left kidney. Got blood and assayed BUN、Cr at 6h、 24h after surgery. Assayed HSP70、 NF-κB by immunohistochemistry and apoptotic cell in left renal tissue by TUNEL.Results: Both perfused dan-shen root group and CsA group decreased BUN、 Gr、 MDA than that of other groups (P<0.01), and increased activity of SOD (PO.Ol). Contrast with other groups, both perfused dan-shen root group and CsA group decreased the cell apoptosis (P<0.01), but there was no significant difference between them (P>0.05). The expression of positive HSP70 in perfused CsA group was higher than that of other groups (P<0.01), and the activity of NF-κB was lower than that of other groups (PO.Ol).Conclusion: The dan-shen root can directly inhibit the injury of OFR to transplanted renal, and CsA can increase the expression of HSP70 to prevent the transplanted renal.
引文
1. Massberg S, Messmer K. The nature of ischemia/reperfusion injury.Transplantation Proceedings,2002,30:4217-4223.
2. Shapiro JI, Cheung C, Itabashi A, et al. The effect of verapamil on renal function after warm and cold ischemia in the isolated perfused rat kidney. Transplant,1985,40:596-599.
3. Fellstrom B. Progression of chonic renal transplant dysfunction. Transplation,2001,33:3355-3356.
4. Szabo A, Heemann U. Ischemia reperfusion injury and chronic allograft rejection. Transplantation Proceedings,1998,30:4281-4284.
5. Azuma H, Nadeau K, Takada M, et al. Cellar and molecular predictors of a single kidney.Transplant,1997,64:192-196.
6. John DH, Samer GM, Chang YC, et al. Renal artery perfusion modifies ischemia/reperfusion injury. Journal of surgical research,1996,60:321-326.
7. Troppmamn C, Gillingham KJ, Benedett IE, et al. Delayed graft function, acute rejection, and outcome after cadaver renal transplantation. Transplant,1995,59:962-963.
8. Yokoyama H, Uchica K, Kobayashi T, et al. Effect of prolonged delayed graft function on long-term outcome in cadaveric kidney transplantation. Clin Transplant,1994,8:101-103.
9. Ibrahim S, Jacobs F, Zukin Y, et al. Immunohistochemical manifestation of unilateral ischemia. Clin Transplantation,1996,10:646-649.
10. Goes N, Urmson J, Ramarssa V, et al. Ischemia acute tubular necrosis induces an extensive local cytokine response evidence for induction of interferon-gamma, transforming growth factor-beta 1, granulocytemacrophage colony-stimulating factor, interleukin-2 and interleukin-10. Tranplant,1995,59:565-567.
11. Djo AO, Wolfe RA, Held PJ, et al. Delayed Graft function: risk factors and implication for renal allograft survival, Transplant,1997,63:969-973.
12. Stoner M, Albadawi H, Cassan P, et al. Poly-ADP ribose polymerase inhibition in a murine model or thoracic aortic ischemia reperfusion: Renal and cardiac benefits. Journal of the American College of Surgeons,2004,199:103-105.
13. Troppmann C, Gruessner AC Gillingham K J, et al. Impact of delayed function on long-term graft survival after solid organ transplantation. Transplantation Proceedings, 1999, 31: 1290-1292.
14. Bonvente JV, Colinvin RB. Adhension molecules in renal disease. Curt Opin Nephrol Hypertens, 1996, 5: 254-256.
15. Anaya PR, Toledo PLH. The molecular events underlying ischemia/reperfusion injury. Transplantation Proceedings, 2002, 34: 2518-2519.
16. Granger DN, Rutili G; Cord JM, et al. Superoxide radicals in intestinal ischemia. Gastroentorology, 1981, 32: 137.
17. Lunec J, Fuller B, Green CJ. Evidence of lipid peroxidation damade in rabbit kidneys during ischemia. Cryobiology. 1983. 20: 731-734.
18.谭建明,欧良明,张仰奎,等.犬肾移植再灌注损伤中自由基的作用.中华泌尿外科杂志,1992,13:23-25.
19.沈寅初,李维才,王明贵,等.丹参对热缺血肾保护作用的实验研究.中华外科杂志,1988,26:759-761.
20. Chumer M, Colombel MC, Sawchuk TS, et al. Morphologic biochemical and molecular evidence of apoptosis during the reperfusion phase after brief periods of renal ischemia. Am J Pathol, 1992, 140: 831-832.
21. Galang N, Sasaki H, Maulik N. Apoptotic cell death during ischemia: reperfusion and its attenuation by antioxidant therpy. Toxicology, 2000, 148: 111-118.
22. Chul WY, Hee JA, Hyuk JH, et al. Pharmacological preconditioning with low-dose cyclosporine or FK506 reduces subsequent ischemia/reperfusion injury in rat kidney. Transplantation, 2001, 72: 1753-1759.
23. Blackwell TS, Christman JW. The role of nuclear factor-κb in cytokine gene regulation. Am J Respir Cell Mol Biol, 1997, 17: 3-4.
24. Juan AC, Nicolai MD, Andrily MB, et al. Opening of potassium channels protects mitochondrial function from calcium overload. Journal of Surgical Research, 2000, 94: 116-123.
25. Roberto AP, Luis HTP, Alex BL, et al. Research review ischemia/reperfusion injury. Journal of Surgical Reseach, 2002, 105: 248-258.
26. Kaur J, Rathan R. Induction of apoptosis by abrogation of HSP70 expression in human and cancer cells. Int J Cancer, 2000, 85: 1-5.
2. Shapiro JI, Cheung C, Itabashi A, et al. The effect of verapamil on renal function after warm and cold ischemia in the isolated perfused rat kidney. Transplant,1985,40:596-599.
3. Fellstrom B. Progression of chonic renal transplant dysfunction. Transplation,2001,33:3355-3356.
4. Szabo A, Heemann U. Ischemia reperfusion injury and chronic allograft rejection. Transplantation Proceedings,1998,30:4281-4284.
5. Azuma H, Nadeau K, Takada M, et al. Cellar and molecular predictors of a single kidney.Transplant,1997,64:192-196.
6. John DH, Samer GM, Chang YC, et al. Renal artery perfusion modifies ischemia/reperfusion injury. Journal of surgical research,1996,60:321-326.
7. Troppmamn C, Gillingham KJ, Benedett IE, et al. Delayed graft function, acute rejection, and outcome after cadaver renal transplantation. Transplant,1995,59:962-963.
8. Yokoyama H, Uchica K, Kobayashi T, et al. Effect of prolonged delayed graft function on long-term outcome in cadaveric kidney transplantation. Clin Transplant,1994,8:101-103.
9. Ibrahim S, Jacobs F, Zukin Y, et al. Immunohistochemical manifestation of unilateral ischemia. Clin Transplantation,1996,10:646-649.
10. Goes N, Urmson J, Ramarssa V, et al. Ischemia acute tubular necrosis induces an extensive local cytokine response evidence for induction of interferon-gamma, transforming growth factor-beta 1, granulocytemacrophage colony-stimulating factor, interleukin-2 and interleukin-10. Tranplant,1995,59:565-567.
11. Djo AO, Wolfe RA, Held PJ, et al. Delayed Graft function: risk factors and implication for renal allograft survival, Transplant,1997,63:969-973.
12. Stoner M, Albadawi H, Cassan P, et al. Poly-ADP ribose polymerase inhibition in a murine model or thoracic aortic ischemia reperfusion: Renal and cardiac benefits. Journal of the American College of Surgeons,2004,199:103-105.
13. Troppmann C, Gruessner AC Gillingham K J, et al. Impact of delayed function on long-term graft survival after solid organ transplantation. Transplantation Proceedings, 1999, 31: 1290-1292.
14. Bonvente JV, Colinvin RB. Adhension molecules in renal disease. Curt Opin Nephrol Hypertens, 1996, 5: 254-256.
15. Anaya PR, Toledo PLH. The molecular events underlying ischemia/reperfusion injury. Transplantation Proceedings, 2002, 34: 2518-2519.
16. Granger DN, Rutili G; Cord JM, et al. Superoxide radicals in intestinal ischemia. Gastroentorology, 1981, 32: 137.
17. Lunec J, Fuller B, Green CJ. Evidence of lipid peroxidation damade in rabbit kidneys during ischemia. Cryobiology. 1983. 20: 731-734.
18.谭建明,欧良明,张仰奎,等.犬肾移植再灌注损伤中自由基的作用.中华泌尿外科杂志,1992,13:23-25.
19.沈寅初,李维才,王明贵,等.丹参对热缺血肾保护作用的实验研究.中华外科杂志,1988,26:759-761.
20. Chumer M, Colombel MC, Sawchuk TS, et al. Morphologic biochemical and molecular evidence of apoptosis during the reperfusion phase after brief periods of renal ischemia. Am J Pathol, 1992, 140: 831-832.
21. Galang N, Sasaki H, Maulik N. Apoptotic cell death during ischemia: reperfusion and its attenuation by antioxidant therpy. Toxicology, 2000, 148: 111-118.
22. Chul WY, Hee JA, Hyuk JH, et al. Pharmacological preconditioning with low-dose cyclosporine or FK506 reduces subsequent ischemia/reperfusion injury in rat kidney. Transplantation, 2001, 72: 1753-1759.
23. Blackwell TS, Christman JW. The role of nuclear factor-κb in cytokine gene regulation. Am J Respir Cell Mol Biol, 1997, 17: 3-4.
24. Juan AC, Nicolai MD, Andrily MB, et al. Opening of potassium channels protects mitochondrial function from calcium overload. Journal of Surgical Research, 2000, 94: 116-123.
25. Roberto AP, Luis HTP, Alex BL, et al. Research review ischemia/reperfusion injury. Journal of Surgical Reseach, 2002, 105: 248-258.
26. Kaur J, Rathan R. Induction of apoptosis by abrogation of HSP70 expression in human and cancer cells. Int J Cancer, 2000, 85: 1-5.