酸中毒收缩大鼠离体冠状动脉机制的研究

发布时间：2018-06-26 00:38

本文选题：酸中毒 + 冠状动脉　；参考：《山西医科大学》2017年硕士论文

【摘要】：目的:观察酸中毒(pH_(ex)6.8)对大鼠离体冠状动脉(coronary artery,CA)静息张力的影响,并探讨其作用机制。通过观察Na+-H+交换体亚型1(NHE-1)抑制剂和Na+-HCO3-共同转运体(NBC)抑制剂对pH_(ex)6.8收缩大鼠离体CA的影响,探讨酸碱转运体在酸中毒引起大鼠离体CA收缩中的作用;通过观察氯通道阻滞剂(NPPB和NFA)及细胞外去除氯离子对pH_(ex)6.8收缩大鼠离体CA的影响,探讨氯离子跨细胞膜转运在酸中毒引起大鼠离体CA收缩中的作用;通过观察Rho激酶(ROCK)抑制剂、蛋白激酶C(PKC)抑制剂和细胞外调节蛋白激酶(ERK)抑制剂对pH_(ex)6.8收缩大鼠离体CA的影响,探讨蛋白激酶在酸中毒引起大鼠离体CA收缩中的作用。方法:1.将SD雄性青年大鼠(230~260 g)断头处死后,快速从胸腔内取出心脏,置于4℃、pH 7.40、95%O_2+5%CO_2饱和的生理盐溶液(Physiological saline solution,PSS)中。在解剖显微镜下,用显微剪和显微镊将大鼠冠状动脉(室壁支、室间隔支及前降支)快速分离干净,游离出来,剪成约2 mm长的冠脉血管环,固定于微血管张力记录仪(Multi Myograph System-610M,DMT)。采用Power Lab微血管环张力系统,记录大鼠离体CA血管环的张力。2.分别观察NHE-1选择性抑制剂HOE-642(30μmol/L)预孵和NBC抑制剂S0859(100μmol/L)预孵对pH_(ex)6.8引起大鼠离体CA收缩的影响。3.分别观察氯离子通道阻滞剂NPPB(10、30、100μmol/L)和NFA(10、30、100μmol/L)对pH_(ex)6.8引起大鼠离体CA收缩的影响;分别观察氯离子通道阻滞剂NPPB(10、30、100、300μmol/L)和NFA(10、30、100、300μmol/L)对KCl(60 mmol/L)引起大鼠离体CA收缩的影响;以及分别观察NPPB(100μmol/L)和NFA(100μmol/L)对血栓素A2类似物U46619(1μmol/L)引起大鼠离体CA收缩的影响。此外,观察用L-天冬氨酸钠等量代替细胞外PSS液中氯化钠后,对pH_(ex)6.8、KCl(60 mmol/L)和U46619(1μmol/L)引起大鼠离体CA收缩的影响。4.分别观察ROCK抑制剂Y-27632(3μmol/L)、PKC抑制剂G?6983(1μmol/L)和ERK抑制剂PD98059(10μmol/L)对pH_(ex)6.8引起大鼠离体CA收缩的影响。结果:1.在静息状态时,pH_(ex)6.8引起大鼠离体CA张力升高,最大张力为(3.90±0.95)m N,相当于KCl(60 mmol/L)最大收缩幅度的(105.07±10.65)%。2.HOE-642(30μmol/L)使pH_(ex)6.8引起的大鼠离体CA收缩幅度降低(18.46±5.29)%(P0.01),S0859(100μmol/L)使其收缩幅度降低(14.90±3.24)%(P0.01)。3.NPPB和NFA均浓度依赖性(10、30、100μmol/L)地抑制pH_(ex)6.8引起的大鼠离体CA收缩,最大抑制百分比分别为(66.61±7.07)%(P0.01)和(64.48±11.68)%(P0.01)。NPPB和NFA均浓度依赖性(10、30、100、300μmol/L)地抑制KCl(60 mmol/L)引起的大鼠离体CA收缩,最大抑制百分比分别为(83.51±3.13)%(P0.01)和(52.37±12.31)%(P0.01)。NPPB(100μmol/L)和NFA(100μmol/L)均可抑制U46619(1μmol/L)引起的大鼠离体CA收缩,其抑制百分比分别为(44.04±9.68)%(P0.01)和(46.23±5.24)%(P0.01)。用L-天冬氨酸钠代替PSS溶液中的Na Cl后,几乎完全抑制pH_(ex)6.8引起的大鼠离体CA收缩(P0.01),而对KCl(60 mmol/L)和U46619(1μmol/L)引起的大鼠离体CA收缩无显著影响(P0.05)。4.Y-27632(3μmol/L)、G?6983(1μmol/L)和PD98059(10μmol/L)均可抑制pH_(ex)6.8引起的大鼠离体CA收缩,其抑制百分比分别为(29.37±13.44)%(P0.01)、(29.84±10.58)%(P0.01)和(23.14±9.47)%(P0.01)。结论:1.酸中毒引起大鼠离体CA收缩与激活NHE-1和NBC有关。2.氯离子跨细胞膜转运和氯离子通道在酸中毒引起大鼠离体CA收缩中起重要作用。3.酸中毒引起大鼠离体CA收缩与激活ROCK、PKC和ERK有关。
[Abstract]:Objective: To observe the effect of acidosis (pH_ (Ex) 6.8) on resting tension of isolated coronary artery (coronary artery (CA)) in rats and explore its mechanism of action. The effect of Na+-H+ exchange body subtype 1 (NHE-1) inhibitor and Na+-HCO3- common transporter (NBC) inhibitor on pH_ (Ex) 6.8 rats' isolated CA was investigated, and the acid base transporter in acidosis was investigated. By observing the effect of chlorine channel blockers (NPPB and NFA) and the effect of chlorine ion removal from pH_ (Ex) 6.8 in vitro CA in vitro, the effect of chloride ion transcellular membrane transport on the contraction of CA in rats in vitro was investigated. By observing the inhibitors of Rho kinase (ROCK), protein kinase C (PKC) inhibitors and CA The effect of extracellular regulated protein kinase (ERK) inhibitor on the isolated CA in pH_ (Ex) 6.8 rat in vitro, and to explore the role of protein kinase in the contraction of CA in rats induced by acidosis. Method: 1. after the death of SD male young rats (230~260 g), the heart was quickly removed from the thoracic cavity and placed at 4, pH 7.40,95%O_2+5%CO_2 saturated physiological salt solution. (Physiological saline solution, PSS). Under the anatomical microscope, the coronary artery (ventricular wall, interventricular septum branch and anterior descending branch) of rats were quickly separated and cleaned by microscissors and microtweezers, free and cut into about 2 mm long coronary vessels, fixed to the microvascular tension recorder (Multi Myograph System-610M, DMT). Power Lab microvessels were used. Cyclic tension system, the tension.2. of CA vascular rings in rats was recorded. The effect of NHE-1 selective inhibitor HOE-642 (30 mu mol/L) pre incubation and NBC inhibitor S0859 (100 micron mol/L) preincubation on pH_ (Ex) 6.8 induced CA contraction in rats was observed, and.3. channel blockers were observed and 6. were observed respectively. 8 the effect of CA contraction in rats in vitro; the effect of chloride channel blocker NPPB (10,30100300 mu mol/L) and NFA (10,30100300 mu mol/L) on KCl (60 mmol/L) induced contraction of rat CA in vitro; and the contraction of thromboxane analogues (1 mu), respectively, caused by NPPB (100 micron) and 100 micron (1 mu) In addition, the effect of sodium chloride on pH_ (Ex) 6.8, KCl (60 mmol/L) and U46619 (1 mu mol/L) induced CA contraction in rats was observed with sodium aspartic acid (L- aspartate). The.4. ROCK inhibitor Y-27632 (3 MU), 6983 (1 mu) and 10 micron (10 mu) for 6.8 were observed. The effect of CA contraction in rat in vitro. Results: 1. in resting state, pH_ (Ex) 6.8 causes the increase of CA tension in rats in vitro, the maximum tension is (3.90 + 0.95) m N, which is equivalent to (105.07 + 10.65)%.2.HOE-642 (30 micron mol/L) of the maximum contraction amplitude of KCl (60 mmol/L), which reduces the contraction amplitude of rats in pH_ (Ex) 6.8 (18.46 + 5.29)% (100 mu). L/L) reduced the contraction amplitude of (14.90 + 3.24)% (P0.01)% (P0.01).3.NPPB and NFA to inhibit pH_ (Ex) 6.8 induced CA contraction in rats in vitro, and the maximum inhibition percentage was (66.61 + 7.07)% (P0.01) and (64.48 + 11.68)% (P0.01). The maximum inhibitory percentage of rat CA in vitro was (83.51 + 3.13)% (P0.01) and (52.37 + 12.31)% (P0.01).NPPB (100 mu mol/L) and NFA (100 mol/L) could inhibit the contraction of rat CA induced by U46619 (1 micron), and the inhibition percentage was (44.04 + 9.68)% (P0.01) and (46.23 + 5.24)% (P0.01), respectively. After Na Cl in SS solution, it almost completely inhibited the CA contraction (P0.01) of rats in vitro caused by pH_ (Ex) 6.8, but there was no significant effect on KCl (60 mmol/L) and U46619 (1 mu mol/L) of rat isolated CA contraction (3 micron). 6983 (1 mu) and 10 micron (10 mu) could inhibit the contraction of rats in vitro. The percentages were (29.37 + 13.44)% (P0.01), (29.84 + 10.58)% (P0.01) and (23.14 + 9.47)% (P0.01). Conclusion: 1. acid poisoning caused CA contraction in rats in vitro and activation of NHE-1 and NBC related to the trans cell membrane transport of.2. chloride ions and the effect of chlorine ion channel on the contraction of CA in rat isolated CA caused by.3. acidosis. Contraction is related to activation of ROCK, PKC and ERK.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R54

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