预适应和前胡丙素对缺氧肥厚血管平滑肌胞内钙离子浓度及No含量的影响
饶曼人 刘宛斌 张晓文
摘 要：观察了预适应和前胡丙素对缺氧肥厚血管平滑肌胞内钙离子浓度([Ca2+]i)及NO含量的影响.①建立两肾一夹肾血管性高血压大鼠模型，分离培养主动脉血管平滑肌细胞(VSMC)，以fura-2/AM为钙指示剂。测得前胡丙素(20mg.kg-1.d-1，术后第9周起ig9周)治疗和缺氧预适应(5minN2，5min95%O2+5%CO2混合气体，循环3次)对缺氧(30minN2)所致肥厚VSMC对KCl和去甲肾上腺素刺激反应性升高([Ca2+]i升高)有明显的拮抗效应.②用血管紧张素Ⅱ刺激致VSMC肥厚，前胡丙素(10μmol.L-1温育24h)和缺氧预适应合用使缺氧肥厚VSMC的NO含量恢复至正常VSMC水平.结果提示前胡丙素与缺氧预适应对肥厚VSMC缺氧损伤有协同性的保护作用.
关键词：前胡丙素;缺血预适应;肌,平滑,血管;细胞,培养的;钙,细胞内;一氧化氮
, 百拇医药 分类号：R972.4 文献标识码：A
文章编号：1000-3002(2001)02-0141-04
Effects of preconditioning and praeruptorine C on intracellular free calcium level and NO content in hypertrophied vascular smooth muscle cells with hypoxia
RAO Man-Ren(Department of Cardiovascular Pharmacology, Nanjing Medical University, Nanjing 210029, China)
LIU Wan-Bing(Department of Cardiovascular Pharmacology, Nanjing Medical University, Nanjing 210029, China)
, 百拇医药
ZHANG Xiao-Wen(Department of Cardiovascular Pharmacology, Nanjing Medical University, Nanjing 210029, China)
Abstract：Effects of preconditioning(PC) and praeruptorine C(Pra-C) on intracellular free calcium level([Ca2+]i) and NO content in hypertrophied vascular smooth muscle cells (VSMC) with hypoxia were studied. ①The aorta VSMC of rats with renovascular hypertension induced by two-kidney-one-clip(2K1C) were isolated and cultured. Fura-2/AM was as a calcium fluorescent indicator. It was seen that Pra-C(20 mg.kg-1.d-1，ig, for 9 weeks from week 9 after 2K1C) and PC (3 cycles of 5 min pure N2 and 5 min 95%O2+5%CO2) antagonisted the elevated activities(higher [Ca2+]i) induced by KCl and norepinephrine in VSMC with hypo- xia (30 min pure N2). ② In hypertrophied VSMC stimulated by angiotensinⅡ, combination of Pra-C(10 μmol.L-1 for 24 h) and PC enhanced NO content to the normal level in hypertrophied VSMC with hypo- xia. The results suggest that combination of Pra-C and PC have cooperative protection for hypertrophied VSMC with hypoxia injury.
, 百拇医药
Keywords：praeruptorine C; preconditioning; muscle, smooth, vascular; cells, cultured; calcium, cytosolic; nitric oxide
基金项目：国家自然科学基金资助项目(39570819)
作者简介：饶曼人(1930-)，女，福建建阳人，教授，心血管药理研究室主任，主要从事心血管药理研究. 联系作者. Tel: (025)6663597,E-mail: Raomanren@263.net
作者单位：饶曼人(南京医科大学心血管药理研究室, 江苏南京 210029)
刘宛斌(南京医科大学心血管药理研究室, 江苏南京 210029)
张晓文(南京医科大学心血管药理研究室, 江苏南京 210029)
, 百拇医药
参考文献：
[1]Murry CE, Jennings RB, Reimer KA. Preconditioning with ischemia: a delay of lethal cell injury in ischemic myocardium[J]. Circulation, 1986, 74(5)：1124-1136.
[2]孙兰, 饶曼人, 刘培庆. 前胡丙素对肾性高血压左室肥厚大鼠心功能, 左室顺应性及心肌胶原含量的影响[J]. 药学学报, 1997, 32(8)：578-582.
[3]王洪新, 饶曼人, 王金唏, 杨思军. 前胡丙素对培养心肌细胞45Ca2+摄入及细胞内游离钙的影响[J]. 中国药理学与毒理学杂志, 1995, 9(3):184-188.
[4]Rao MR, Shen XH, Zou X. Effects of praeruptorin C and E isolated from Qian-Hu on swine and guinea-pig coronary arteria[J]. Eur J Pharmacol, 1988, 155:293-296.
, 百拇医药
[5]孙兰, 饶曼人, 王金唏. 前胡丙素对分离的成年鼠正常及肥厚心室肌细胞内游离钙的影响[J]. 中国药理学报, 1997, 18(3)：251-254.
[6]吴冬梅, 李庆平, 饶曼人. 前胡丙素对肾型高血压大鼠血压及尾动脉反应性的影响[J]. 中国药理学通报, 1997, 13(3)：242-243.
[7]丁小凌, 李云霞. 大鼠肾血管性高血压心肌肥厚的产生和逆转[J]. 湖南医科大学学报, 1989, 14(3)：209-212.
[8]Chamley-Campbell J, Campbell GR, Ross R. The smooth muscle cell in culture[J]. Physiol Rev, 1979, 59(1):1-61.
[9]Williford DJ, Sharma VK, Korth M, Sheu SS. Spatial heterogeneity of intracellular Ca2+ concentration in nonbeating guinea pig ventricular myocytes[J]. Circ Res, 1990, 66(1):241-248.
, 百拇医药
[10]Grynkiewicz G, Poenie M, Tsien RY. A new generation of Ca2+ indicators with greatly improved fluorescence properties[J]. J Biol Chem, 1985, 260(6):3440-3450.
[11]Green LC, Wagner DA, Glogowski J, Skipper PL, Wishnok JS, Tannenbaum SR. Analysis of nitrate, nitrite, and 15Nnitrate in biological fluids[J]. Anal Biochem, 1982, 126(1):131-138.
[12]宋秋景, 李元建. 缺血预适应的研究现状和前景[J]. 中国药理学与毒理学杂志, 2000, 14(1)：1-7.
[13]Massoudy P, Becker BF,Gerlach E. Nitric oxide accounts for postischemic cardioprotection resulting from angiotensin-coverting enzyme inhibition: indirect evidence for a radical scavenger effect in isolated guinea pig heart[J]. J Cardiovasc Pharmacol, 1995, 25(3):440-447., 百拇医药
摘 要：观察了预适应和前胡丙素对缺氧肥厚血管平滑肌胞内钙离子浓度([Ca2+]i)及NO含量的影响.①建立两肾一夹肾血管性高血压大鼠模型，分离培养主动脉血管平滑肌细胞(VSMC)，以fura-2/AM为钙指示剂。测得前胡丙素(20mg.kg-1.d-1，术后第9周起ig9周)治疗和缺氧预适应(5minN2，5min95%O2+5%CO2混合气体，循环3次)对缺氧(30minN2)所致肥厚VSMC对KCl和去甲肾上腺素刺激反应性升高([Ca2+]i升高)有明显的拮抗效应.②用血管紧张素Ⅱ刺激致VSMC肥厚，前胡丙素(10μmol.L-1温育24h)和缺氧预适应合用使缺氧肥厚VSMC的NO含量恢复至正常VSMC水平.结果提示前胡丙素与缺氧预适应对肥厚VSMC缺氧损伤有协同性的保护作用.
关键词：前胡丙素;缺血预适应;肌,平滑,血管;细胞,培养的;钙,细胞内;一氧化氮
, 百拇医药 分类号：R972.4 文献标识码：A
文章编号：1000-3002(2001)02-0141-04
Effects of preconditioning and praeruptorine C on intracellular free calcium level and NO content in hypertrophied vascular smooth muscle cells with hypoxia
RAO Man-Ren(Department of Cardiovascular Pharmacology, Nanjing Medical University, Nanjing 210029, China)
LIU Wan-Bing(Department of Cardiovascular Pharmacology, Nanjing Medical University, Nanjing 210029, China)
, 百拇医药
ZHANG Xiao-Wen(Department of Cardiovascular Pharmacology, Nanjing Medical University, Nanjing 210029, China)
Abstract：Effects of preconditioning(PC) and praeruptorine C(Pra-C) on intracellular free calcium level([Ca2+]i) and NO content in hypertrophied vascular smooth muscle cells (VSMC) with hypoxia were studied. ①The aorta VSMC of rats with renovascular hypertension induced by two-kidney-one-clip(2K1C) were isolated and cultured. Fura-2/AM was as a calcium fluorescent indicator. It was seen that Pra-C(20 mg.kg-1.d-1，ig, for 9 weeks from week 9 after 2K1C) and PC (3 cycles of 5 min pure N2 and 5 min 95%O2+5%CO2) antagonisted the elevated activities(higher [Ca2+]i) induced by KCl and norepinephrine in VSMC with hypo- xia (30 min pure N2). ② In hypertrophied VSMC stimulated by angiotensinⅡ, combination of Pra-C(10 μmol.L-1 for 24 h) and PC enhanced NO content to the normal level in hypertrophied VSMC with hypo- xia. The results suggest that combination of Pra-C and PC have cooperative protection for hypertrophied VSMC with hypoxia injury.
, 百拇医药
Keywords：praeruptorine C; preconditioning; muscle, smooth, vascular; cells, cultured; calcium, cytosolic; nitric oxide
基金项目：国家自然科学基金资助项目(39570819)
作者简介：饶曼人(1930-)，女，福建建阳人，教授，心血管药理研究室主任，主要从事心血管药理研究. 联系作者. Tel: (025)6663597,E-mail: Raomanren@263.net
作者单位：饶曼人(南京医科大学心血管药理研究室, 江苏南京 210029)
刘宛斌(南京医科大学心血管药理研究室, 江苏南京 210029)
张晓文(南京医科大学心血管药理研究室, 江苏南京 210029)
, 百拇医药
参考文献：
[1]Murry CE, Jennings RB, Reimer KA. Preconditioning with ischemia: a delay of lethal cell injury in ischemic myocardium[J]. Circulation, 1986, 74(5)：1124-1136.
[2]孙兰, 饶曼人, 刘培庆. 前胡丙素对肾性高血压左室肥厚大鼠心功能, 左室顺应性及心肌胶原含量的影响[J]. 药学学报, 1997, 32(8)：578-582.
[3]王洪新, 饶曼人, 王金唏, 杨思军. 前胡丙素对培养心肌细胞45Ca2+摄入及细胞内游离钙的影响[J]. 中国药理学与毒理学杂志, 1995, 9(3):184-188.
[4]Rao MR, Shen XH, Zou X. Effects of praeruptorin C and E isolated from Qian-Hu on swine and guinea-pig coronary arteria[J]. Eur J Pharmacol, 1988, 155:293-296.
, 百拇医药
[5]孙兰, 饶曼人, 王金唏. 前胡丙素对分离的成年鼠正常及肥厚心室肌细胞内游离钙的影响[J]. 中国药理学报, 1997, 18(3)：251-254.
[6]吴冬梅, 李庆平, 饶曼人. 前胡丙素对肾型高血压大鼠血压及尾动脉反应性的影响[J]. 中国药理学通报, 1997, 13(3)：242-243.
[7]丁小凌, 李云霞. 大鼠肾血管性高血压心肌肥厚的产生和逆转[J]. 湖南医科大学学报, 1989, 14(3)：209-212.
[8]Chamley-Campbell J, Campbell GR, Ross R. The smooth muscle cell in culture[J]. Physiol Rev, 1979, 59(1):1-61.
[9]Williford DJ, Sharma VK, Korth M, Sheu SS. Spatial heterogeneity of intracellular Ca2+ concentration in nonbeating guinea pig ventricular myocytes[J]. Circ Res, 1990, 66(1):241-248.
, 百拇医药
[10]Grynkiewicz G, Poenie M, Tsien RY. A new generation of Ca2+ indicators with greatly improved fluorescence properties[J]. J Biol Chem, 1985, 260(6):3440-3450.
[11]Green LC, Wagner DA, Glogowski J, Skipper PL, Wishnok JS, Tannenbaum SR. Analysis of nitrate, nitrite, and 15Nnitrate in biological fluids[J]. Anal Biochem, 1982, 126(1):131-138.
[12]宋秋景, 李元建. 缺血预适应的研究现状和前景[J]. 中国药理学与毒理学杂志, 2000, 14(1)：1-7.
[13]Massoudy P, Becker BF,Gerlach E. Nitric oxide accounts for postischemic cardioprotection resulting from angiotensin-coverting enzyme inhibition: indirect evidence for a radical scavenger effect in isolated guinea pig heart[J]. J Cardiovasc Pharmacol, 1995, 25(3):440-447., 百拇医药
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