异丙酚、氯胺酮对兔气管平滑肌细胞内钙离子的影响
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摘要
目的:异丙酚(Propofol)和氯胺酮(Ketamine)作为临床静脉麻醉药,目前广泛应用于临床麻醉。已有研究采用电生理、分子生物及形态学等多项技术,对犬等动物和人的器官和组织从不同角度进行研究,研究表明异丙酚和氯胺酮对气管平滑肌均有不同程度的舒张作用,对气道起到了一定的保护作用。并认为异丙酚的这种保护作用主要与抑制迷走神经张力以及直接扩张平滑肌有关。氯胺酮舒张气管平滑肌的保护作用则可能和抑制ROC,内钙释放或PKC信号系统以及剂量依赖性开放钾离子通道有关。对于二者是否作用于细胞内钙通道以及是否通过内质网起作用,目前国内外文献报道较少。本实验利用激光共聚焦显微镜测定细胞浆内Ca 2+荧光强度峰值的改变,从而探讨不同浓度的异丙酚和氯胺酮对兔气管平滑肌细胞内钙和内质网释放钙的影响,进一步明确其舒张气管平滑肌的作用机制。
方法:实验动物选择新西兰兔,雌雄不拘,体重1.5±0.3㎏。采用酶解方法获得兔的气管平滑肌细胞。选用细胞膜完整、有立体感、梭形、折光好的细胞进行荧光标记。将分离的气管平滑肌细胞离心洗涤,震荡成细胞悬液,加入5μM的FLuo-3/AM后,将细胞悬液放进37℃恒温水浴箱避光孵育30min后,于激光共聚焦显微镜下观察气管平滑肌细胞被标记情况。
实验分组1.异丙酚将急性分离的气管平滑肌细胞随机分为:异丙酚组(Pro组),异丙酚+2-APB组(Pro+2-APB组)和异丙酚+ryanodine组(Pro+RyD组),每组又根据加入异丙酚浓度的不同,再将细胞随机分为:0,30,300μM异丙酚亚组。2.氯胺酮将急性分离的气管平滑肌细胞随机分为:氯胺酮组(Ket组),氯胺酮+2-APB组(Ket+2-APB组)和氯胺酮+ryanodine组(Ket+RyD组),每组又根据加入氯胺酮浓度的不同,再将细胞随机分为:0,100,1000μM氯胺酮亚组。
实验流程
异丙酚组(Pro组)在已用Fluo-3标记好的细胞悬液中,加入终浓度1μM的ACH,记录细胞内钙离子荧光强度峰值。用无钙的HBSS液冲洗后,分别加入终浓度为0μM(加HBSS液),30μM和300μM的异丙酚,孵育15min后,再加入终浓度1μM的ACH,记录各组细胞内钙离子荧光强度峰值。
异丙酚+2-APB组(Pro+2-APB组)在已用Fluo-3标记好的细胞悬液中,加入终浓度1μM的ACH,记录细胞内钙离子荧光强度峰值。加入终浓度40μM的2-APB孵育15min后,分别加入终浓度为0μM(加HBSS液),30μM和300μM的异丙酚,与2-APB共同孵育15min后,用无钙的HBSS液冲洗三次,再加入终浓度1μM的ACH,记录各组细胞内钙离子荧光强度峰值。
异丙酚+RyD组(Pro+RyD组)在已用Fluo-3标记好的细胞悬液中,加入终浓度1μM的ACH,记录细胞内钙离子荧光强度峰值。用无钙的HBSS液冲洗后,加入终浓度10μM的RyD孵育15min后,分别加入终浓度为0μM(加HBSS液),30μM和300μM的异丙酚,与RyD共同孵育15min后,用无钙的HBSS液冲洗三次,再加入终浓度1μM的ACH,记录各组细胞内钙离子荧光强度峰值。
氯胺酮组(Ket组),Ket+2-APB组和Ket+RyD组中,氯胺酮各亚组浓度分别为:0μM,100μM和1000μM。实验流程方法同异丙酚各对应组。结果:1用急性酶分离方法获得的细胞种类单一,易于分辨,分离细胞数量多,成活率高,80%～90%以上的细胞台盼蓝染色鉴定为活细胞,其细胞膜完整、有立体感、梭形、折光性好。细胞形态多种多样,以蚯蚓状和香蕉状多见,占70%～80%,其他还有椭圆形、圆形及不规则形。细胞大小不一,直径12～16vm,长度约60～100vm。细胞轮廓清晰,膜表面光滑,胞内颗粒少。
2与0μM异丙酚组比较,300μM异丙酚组细胞内钙离子荧光强度峰值明显降低(P<0.05)。
3 Pro+2-APB共同作用后,与相应浓度0μM,30μM和300μM异丙酚组(Pro组)比较,0μM与30μM Pro+2-APB组细胞内钙离子的荧光强度峰值均有明显降低(P<0.05)。300μM Pro+2-APB组细胞内钙离子荧光强度峰值无明显降低(P>0.05)。
4 Pro+RyD共同作用后,与相应浓度0μM,30μM和300μM异丙酚组(Pro组)比较,相应浓度0μM,30μM和300μM Pro+RYD组细胞内钙离子荧光强度峰值均有明显降低(P<0.05),与0μM Pro+RYD组比较,300μM Pro+RYD组细胞内钙荧光强度峰值明显降低(P<0.05)。
5与0μM氯胺酮组比较,1000μM氯胺酮组细胞内钙离子荧光强度
峰值明显降低(P<0.05)。6 Ket+2-APB共同作用后,与氯胺酮组(Ket组)中的各对应浓度组比较,0μM与100μM Ket+2-APB组的细胞内钙离子的荧光强度峰值均有明显降低(P<0.05)。1000μM Ket+2-APB组细胞内钙离子荧光强度峰值无明显变化(P>0.05)。
7 Ket+RyD共同作用后,与氯胺酮组(Ket组)中的各对应浓度组比较,0μM、100μM与1000μM Ket+RyD组细胞内钙离子荧光强度峰值均有明显降低(P<0.05)。与0μM Ket+RyD组比较,1000μM Ket+RyD组细胞内钙离子荧光强度峰值有明显降低(P<0.05)。
结论:1 30μM异丙酚对细胞内钙无影响,300μM异丙酚可以明显降低气管平滑肌细胞内钙。
2异丙酚降低细胞内钙可能与其抑制内质网IP3通路有关,与内质网ryanodine通路无关。
3 100μM氯胺酮对细胞内钙无影响。1000μM氯胺酮可以明显降低气管平滑肌细胞内钙。
4氯胺酮可能抑制内质网上的IP3通路,抑制内质网内的钙释放,从而降低胞浆内钙浓度,而可能与内质网ryanodine通道无关。
Objective: Propofol and Ketamine are generally used for clinical anesthesia. Studies show that Propofol and Ketamine have different degree relaxation on tracheal smooth muscles, through which way to protect the airway. Propofol reacts mainly through inhibiting the cranial nerve tension; while in high concentrations, it has direct relaxing action on tracheal smooth muscles. Ketamine relaxes tracheal smooth muscles possibly concerned with inhibition of ROC, delivery of intracellular calcium PKC signal system, or dose dependent patency of kalium ion channel. There are less reports about whether the two have effects on intracellular calcium and react through endoplasmic reticulum. Our experiments approach the effects of different concentrations of Propofol and Ketamine on intracellular calcium and releasing of endoplasmic reticulum calcium through evaluating the variation of intracytoplasm calcium; and further identify their mechanisms of action on relaxing tracheal smooth muscles.
Methods: New Zealand rabbit are chosen as experimental animals, no limits to their sex, and 1.5±0.3kg weight. The single rabbit tracheal smooth muscle cells are got by enzymolysis methods. Choose cells with complete cell membrane, three-dimensional, fusiform and well refraction function to begin fluorescent labeling. Shock the centrifuged cells into cell suspension, and put 5μM concentration FLuo-3/AM into the cell suspension. Incubate cell suspension in thermostatic waterbath box under 37℃for 30 min. Observe the labeling conditions of the trachea smooth muscle cells under the laser confocal microscopy.
Groups 1.propofol Fractionate the cell suspension treated by the methods above into Propofol group, Pro+2-APB group and Pro+ryanodine group(Pro+RyD); and each group is fractionated into three subsets according to the density of propofol, that is 0, 30 and 300μM group. 2.ketamine Fractionate the cell suspension treated by the methods above into ketamine group ,ket+ 2-APB group and ket+ RyD group, and each group is fractionated into three subsets according to the density of ketamine, that is 0, 100 and 1000μM group.
Experiment flow-sheet
Propofol group(Cgroup) add 1μM ACH to the cell suspension signatured by FLuo-3, and record the calcium fluorescence peak intensity. Wash cell suspension with HBSS for 3 times. Then add different density propofol into each subset groups to incubate for 15 min; add 1μM ACH again . Record the calcium fluorescence peak intensity in each group.
Pro+2-APB group add 1μM ACH to the cell suspension signatured by FLuo-3. Record the calcium fluorescence peak intensity. Then add 40μM 2-APB; incubate for 15 min; then add different density propofol respectively , and incubate with 2-APB together for 15 min; wash with HBSS for 3 times, add 1μM ACH again .Record the calcium fluorescence peak intensity in each group.
Pro+RyD group add 1μM ACH to the cell suspension signatured by FLuo-3. Record the calcium fluorescence peak intensity. Wash cell suspension with HBSS for 3 times. Then add 10μM RyD incubate for 15 min; then add different density propofol; incubate with RyD together for 15 min; wash with HBSS for 3 times, add 1μM ACH again . Record the calcium fluorescence peak intensity in each group.
In ketamine group(C group), ket+2-APB group and ket+RyD group, the density of ketamine is 0,100and 1000μM. The methods are the same to propofol.
Results: 1 Cells aquired through enzyme separation method are pure, easy to discern, large quantity, and high survival rate. 80%~90% of the cells are identified as living cells by trypan blue dyeing method. The cell appearances are varied, well about 70%~80% of them are just like the earthworms and bananas; also other shapes as ellipse, round and irregularity. Sizes of the cells are not at equal. The average diameter of them is about 12~16vm, and the length about 60~100vm. The edge of the cells is distinct; the membrane surface is smooth; endoparticle is few.
2 Compared with 0μM propofol group, 300μM propofol can decrease the intracellular calcium peak intensity obviously (P<0.05).
3 After treated by Pro+2-APB, compared with their corresponding groups in the control group , there are obvious decrease in 0μM and 30μM Pro+2-APB group(P<0.05); and no differences in 300μM Pro+2-APB group(P>0.05).
4 After treated by Pro+RyD, compared with their corresponding groups in the control group , the intracellular calcium peak intensity in each group has an obvious decrease(P<0.05). Compared with 0μM Pro+RyD group , the intracellular calcium peak intensity decreased obviously in 300μM Pro+RyD group(P<0.05).
5 Compared with 0μM ketamine group , there is obvious decrease in 1000μM ketamine group(P<0.05).
6 After treated by ket+2-APB, Compared with their corresponding groups in the control group, the intracellular calcium peak intensity in 0μM and 100μM ketamine+2-APB groups have obvious reduction(P<0.05) . No obvious decrease in 1000μM ketamine+2-APB group(P>0.05).
7 After treated by ket+RyD, compared with their corresponding groups in the control group, the intracellular calcium peak intensity have obvious decrease in each ket+RyD group(P<0.05); compared with 0μM ket+RyD group, 1000μM ket+RyD group has obvious decrease(P<0.05) . Conclusions: 1 300μM propofol can decrease the intracellular calcium, while 30μM cannot.
2 Propofol maybe block the IP3 way to decrease the intracellular calcium ,while no relationship with ryanodine way.
3 1000μM ketamine can decrease the intracellular calcium, while 100μM cannot.
4 Ketamine could block the IP3 way to inhibit the calcium release from endoplasmic reticulum to decrease intracellular calcium .while the ryanodine way cannot be blocked by ketamine.
方法:实验动物选择新西兰兔,雌雄不拘,体重1.5±0.3㎏。采用酶解方法获得兔的气管平滑肌细胞。选用细胞膜完整、有立体感、梭形、折光好的细胞进行荧光标记。将分离的气管平滑肌细胞离心洗涤,震荡成细胞悬液,加入5μM的FLuo-3/AM后,将细胞悬液放进37℃恒温水浴箱避光孵育30min后,于激光共聚焦显微镜下观察气管平滑肌细胞被标记情况。
实验分组1.异丙酚将急性分离的气管平滑肌细胞随机分为:异丙酚组(Pro组),异丙酚+2-APB组(Pro+2-APB组)和异丙酚+ryanodine组(Pro+RyD组),每组又根据加入异丙酚浓度的不同,再将细胞随机分为:0,30,300μM异丙酚亚组。2.氯胺酮将急性分离的气管平滑肌细胞随机分为:氯胺酮组(Ket组),氯胺酮+2-APB组(Ket+2-APB组)和氯胺酮+ryanodine组(Ket+RyD组),每组又根据加入氯胺酮浓度的不同,再将细胞随机分为:0,100,1000μM氯胺酮亚组。
实验流程
异丙酚组(Pro组)在已用Fluo-3标记好的细胞悬液中,加入终浓度1μM的ACH,记录细胞内钙离子荧光强度峰值。用无钙的HBSS液冲洗后,分别加入终浓度为0μM(加HBSS液),30μM和300μM的异丙酚,孵育15min后,再加入终浓度1μM的ACH,记录各组细胞内钙离子荧光强度峰值。
异丙酚+2-APB组(Pro+2-APB组)在已用Fluo-3标记好的细胞悬液中,加入终浓度1μM的ACH,记录细胞内钙离子荧光强度峰值。加入终浓度40μM的2-APB孵育15min后,分别加入终浓度为0μM(加HBSS液),30μM和300μM的异丙酚,与2-APB共同孵育15min后,用无钙的HBSS液冲洗三次,再加入终浓度1μM的ACH,记录各组细胞内钙离子荧光强度峰值。
异丙酚+RyD组(Pro+RyD组)在已用Fluo-3标记好的细胞悬液中,加入终浓度1μM的ACH,记录细胞内钙离子荧光强度峰值。用无钙的HBSS液冲洗后,加入终浓度10μM的RyD孵育15min后,分别加入终浓度为0μM(加HBSS液),30μM和300μM的异丙酚,与RyD共同孵育15min后,用无钙的HBSS液冲洗三次,再加入终浓度1μM的ACH,记录各组细胞内钙离子荧光强度峰值。
氯胺酮组(Ket组),Ket+2-APB组和Ket+RyD组中,氯胺酮各亚组浓度分别为:0μM,100μM和1000μM。实验流程方法同异丙酚各对应组。结果:1用急性酶分离方法获得的细胞种类单一,易于分辨,分离细胞数量多,成活率高,80%～90%以上的细胞台盼蓝染色鉴定为活细胞,其细胞膜完整、有立体感、梭形、折光性好。细胞形态多种多样,以蚯蚓状和香蕉状多见,占70%～80%,其他还有椭圆形、圆形及不规则形。细胞大小不一,直径12～16vm,长度约60～100vm。细胞轮廓清晰,膜表面光滑,胞内颗粒少。
2与0μM异丙酚组比较,300μM异丙酚组细胞内钙离子荧光强度峰值明显降低(P<0.05)。
3 Pro+2-APB共同作用后,与相应浓度0μM,30μM和300μM异丙酚组(Pro组)比较,0μM与30μM Pro+2-APB组细胞内钙离子的荧光强度峰值均有明显降低(P<0.05)。300μM Pro+2-APB组细胞内钙离子荧光强度峰值无明显降低(P>0.05)。
4 Pro+RyD共同作用后,与相应浓度0μM,30μM和300μM异丙酚组(Pro组)比较,相应浓度0μM,30μM和300μM Pro+RYD组细胞内钙离子荧光强度峰值均有明显降低(P<0.05),与0μM Pro+RYD组比较,300μM Pro+RYD组细胞内钙荧光强度峰值明显降低(P<0.05)。
5与0μM氯胺酮组比较,1000μM氯胺酮组细胞内钙离子荧光强度
峰值明显降低(P<0.05)。6 Ket+2-APB共同作用后,与氯胺酮组(Ket组)中的各对应浓度组比较,0μM与100μM Ket+2-APB组的细胞内钙离子的荧光强度峰值均有明显降低(P<0.05)。1000μM Ket+2-APB组细胞内钙离子荧光强度峰值无明显变化(P>0.05)。
7 Ket+RyD共同作用后,与氯胺酮组(Ket组)中的各对应浓度组比较,0μM、100μM与1000μM Ket+RyD组细胞内钙离子荧光强度峰值均有明显降低(P<0.05)。与0μM Ket+RyD组比较,1000μM Ket+RyD组细胞内钙离子荧光强度峰值有明显降低(P<0.05)。
结论:1 30μM异丙酚对细胞内钙无影响,300μM异丙酚可以明显降低气管平滑肌细胞内钙。
2异丙酚降低细胞内钙可能与其抑制内质网IP3通路有关,与内质网ryanodine通路无关。
3 100μM氯胺酮对细胞内钙无影响。1000μM氯胺酮可以明显降低气管平滑肌细胞内钙。
4氯胺酮可能抑制内质网上的IP3通路,抑制内质网内的钙释放,从而降低胞浆内钙浓度,而可能与内质网ryanodine通道无关。
Objective: Propofol and Ketamine are generally used for clinical anesthesia. Studies show that Propofol and Ketamine have different degree relaxation on tracheal smooth muscles, through which way to protect the airway. Propofol reacts mainly through inhibiting the cranial nerve tension; while in high concentrations, it has direct relaxing action on tracheal smooth muscles. Ketamine relaxes tracheal smooth muscles possibly concerned with inhibition of ROC, delivery of intracellular calcium PKC signal system, or dose dependent patency of kalium ion channel. There are less reports about whether the two have effects on intracellular calcium and react through endoplasmic reticulum. Our experiments approach the effects of different concentrations of Propofol and Ketamine on intracellular calcium and releasing of endoplasmic reticulum calcium through evaluating the variation of intracytoplasm calcium; and further identify their mechanisms of action on relaxing tracheal smooth muscles.
Methods: New Zealand rabbit are chosen as experimental animals, no limits to their sex, and 1.5±0.3kg weight. The single rabbit tracheal smooth muscle cells are got by enzymolysis methods. Choose cells with complete cell membrane, three-dimensional, fusiform and well refraction function to begin fluorescent labeling. Shock the centrifuged cells into cell suspension, and put 5μM concentration FLuo-3/AM into the cell suspension. Incubate cell suspension in thermostatic waterbath box under 37℃for 30 min. Observe the labeling conditions of the trachea smooth muscle cells under the laser confocal microscopy.
Groups 1.propofol Fractionate the cell suspension treated by the methods above into Propofol group, Pro+2-APB group and Pro+ryanodine group(Pro+RyD); and each group is fractionated into three subsets according to the density of propofol, that is 0, 30 and 300μM group. 2.ketamine Fractionate the cell suspension treated by the methods above into ketamine group ,ket+ 2-APB group and ket+ RyD group, and each group is fractionated into three subsets according to the density of ketamine, that is 0, 100 and 1000μM group.
Experiment flow-sheet
Propofol group(Cgroup) add 1μM ACH to the cell suspension signatured by FLuo-3, and record the calcium fluorescence peak intensity. Wash cell suspension with HBSS for 3 times. Then add different density propofol into each subset groups to incubate for 15 min; add 1μM ACH again . Record the calcium fluorescence peak intensity in each group.
Pro+2-APB group add 1μM ACH to the cell suspension signatured by FLuo-3. Record the calcium fluorescence peak intensity. Then add 40μM 2-APB; incubate for 15 min; then add different density propofol respectively , and incubate with 2-APB together for 15 min; wash with HBSS for 3 times, add 1μM ACH again .Record the calcium fluorescence peak intensity in each group.
Pro+RyD group add 1μM ACH to the cell suspension signatured by FLuo-3. Record the calcium fluorescence peak intensity. Wash cell suspension with HBSS for 3 times. Then add 10μM RyD incubate for 15 min; then add different density propofol; incubate with RyD together for 15 min; wash with HBSS for 3 times, add 1μM ACH again . Record the calcium fluorescence peak intensity in each group.
In ketamine group(C group), ket+2-APB group and ket+RyD group, the density of ketamine is 0,100and 1000μM. The methods are the same to propofol.
Results: 1 Cells aquired through enzyme separation method are pure, easy to discern, large quantity, and high survival rate. 80%~90% of the cells are identified as living cells by trypan blue dyeing method. The cell appearances are varied, well about 70%~80% of them are just like the earthworms and bananas; also other shapes as ellipse, round and irregularity. Sizes of the cells are not at equal. The average diameter of them is about 12~16vm, and the length about 60~100vm. The edge of the cells is distinct; the membrane surface is smooth; endoparticle is few.
2 Compared with 0μM propofol group, 300μM propofol can decrease the intracellular calcium peak intensity obviously (P<0.05).
3 After treated by Pro+2-APB, compared with their corresponding groups in the control group , there are obvious decrease in 0μM and 30μM Pro+2-APB group(P<0.05); and no differences in 300μM Pro+2-APB group(P>0.05).
4 After treated by Pro+RyD, compared with their corresponding groups in the control group , the intracellular calcium peak intensity in each group has an obvious decrease(P<0.05). Compared with 0μM Pro+RyD group , the intracellular calcium peak intensity decreased obviously in 300μM Pro+RyD group(P<0.05).
5 Compared with 0μM ketamine group , there is obvious decrease in 1000μM ketamine group(P<0.05).
6 After treated by ket+2-APB, Compared with their corresponding groups in the control group, the intracellular calcium peak intensity in 0μM and 100μM ketamine+2-APB groups have obvious reduction(P<0.05) . No obvious decrease in 1000μM ketamine+2-APB group(P>0.05).
7 After treated by ket+RyD, compared with their corresponding groups in the control group, the intracellular calcium peak intensity have obvious decrease in each ket+RyD group(P<0.05); compared with 0μM ket+RyD group, 1000μM ket+RyD group has obvious decrease(P<0.05) . Conclusions: 1 300μM propofol can decrease the intracellular calcium, while 30μM cannot.
2 Propofol maybe block the IP3 way to decrease the intracellular calcium ,while no relationship with ryanodine way.
3 1000μM ketamine can decrease the intracellular calcium, while 100μM cannot.
4 Ketamine could block the IP3 way to inhibit the calcium release from endoplasmic reticulum to decrease intracellular calcium .while the ryanodine way cannot be blocked by ketamine.
引文
1卢中举,唐大椿,向继洲.平滑肌收缩调节的信号传导.生理科学进展,1997,28:337-340
2孙大业,郭艳林,马力耕.细胞信号转导.生理科学进展,1998.52-168
3 Pan BX, Zhao GL,Huang XL,Zhao KS.Calcium mobilization is required for peroxynitrite-mediated enhancement of spontaneous transient outward currents in arteriolar smooth muscle cells. Free Radic Biol Med 2004,37(6): 823-838
4 Fellner SK,Parker LA.Endothelin B receptor Ca2+ signaling in shark vascular smooth muscle: participation of inositol trisphosphate and ryanodine receptors.J Exp Biol,2004;207(19):3411-3417
5 Ouedraogo N,Roux E,Forestier F,et al.Effects of intravenous anesthetics on normal and passively sensitized human isolated airway smooth muscle. Anesthesiology.1998,88(2):317-26
6刘宝刚,曾帮雄.氯胺酮对主肺动脉平滑肌细胞膜钙通道的影响。中华麻醉学杂志,1997,17(7):427
7 Susan K,Fellen,Willam J.Voltage-gated Ca2+ entry and ryanodine receptor Ca2+-induced Ca2+ release in preglomerular arterioles.America Physiology, 2007,290(5)
8 Yamakage M,Hirshman CA,Croxton TL.Inhibitory effects of thiopental, ketamine,and propofol on voltage-dependent Ca2+channels in porcine tracheal muscle cells.Anesthesiology,1995,83:1274-1281
9聂宏光,崔湧，韩冬云.氯胺酮对哮喘大鼠气管平滑肌细胞ATP敏感钾电流的影响。中国药理学通报,2003,19(12):1381-1384
10 Sinner B,Friedrich O,Zink W,et al.Ketamine stereoselectively inhibits spontaneous Ca2+-oscillations in cultured hippocampal neurons.Anesth Analg, 2005,100:1660-1666
11 Hardingham GE,Chawla S,Johnson CM, et al. Distinct functions of nuclear and cytoplasmic calcium in the control of gene expression.Nature, 1997,385:260-265
12肖欣荣,陈文彬,程德云.慢性缺氧抑制肺动脉平滑肌细胞钾通道活性[J].中华结核和呼吸杂志,1998,21(11):681-5
13刘杰,赵克森,金春华.重症休克时细动脉平滑肌细胞膜ATP敏感钾通道的变化[J].中华创伤杂志,1998,14(增刊1):10-12
14 Szabadkai G,Rizzuto R. Participation of endoplasmic reticulum and mitochondrial calcium handling in apoptosis:more than just neighborhood [J].FEBSLett,2004,567(1):111-115
15 Verkhratsky A,Toescu EC.Endoplasm in reticulum Ca2+ homeostasis and neuronal death[J].J Cell Mol Med,2003,7(4):351-361
16 Bardo S,Cavazzini MG, Emptage N.The role of the endoplasmic reticulum Ca2+store in the plasticity of central neurons[J].Trends Pharmacol Sci, 2006,27(2):78-84
17卢中举,唐大椿,向继洲.平滑肌收缩调节的信号传导.生理科学进展,1997,28:337-340
18孙大业,郭艳林,马力耕.细胞信号转导.生理科学进展,1998.52-168
19 Missiaen L,Callewaert G,De Smedt H,Parys JB.2-Aminoethoxydiphenyl borate affects the inositol 1,4,5-trisphosphate receptor, the intracellular Ca2+ pump and the non-specific Ca2+ leak from the non-mitochondrial Ca2+ stores in permeabilized cells.Cell Calcium 2001;29(2):111-116
20 Gordienko DV,Bolton TB.Crosstalk between ryanodine receptors and IP3 receptors as a factor shaping spontaneous Ca2+-release events in rabbit portal vein myocytes.J Physiol 2002;542(3): 743-762
21 Susan K,Fellen,Willam J. Voltage-gated Ca2+ entry and ryanodine receptor Ca2+-induced Ca2+ release in preglomerular arterioles.America Physiology, 2007,290(5)
22 Yamakage M, Kohro S, Kawamata T, et al. Inhibitory effects of four inhaled anesthetics on canine tracheal smooth muscle contraction and intracelluar Ca2+ concentration.Anesth Analg,1993,77:67-72
23 liN,YinL,SuZL.Laser Scanning Confocol Microtechnique[M].Beijing: Renmin Military Medical Publishing House,1997,61-64
24魏敏杰,李智,王俊科等.异丙酚舒张血管平滑肌的作用机制.中华麻醉学杂志,1997,17:676-679
25 Cheng EY,Mazzeo AJ,Bosniak ZJ etal.Direct relaxant effects of intravenous anesthetics on airway smooth muscle.Anesth Analg, 1996, 83:162- 168
26 Mehr EH,Lindeman KS.Effects of halothane,propofol,and thiopental on peripheral airway reactivity.Anesthesiology,1993,79:290-298
27 Conti G,Dell’Utri D,Uilardi V,etal.Propofol induces bronchodilation in mechanically ventilated chronic obstructive pulmonary disease(COPD) patients.Acta Anesthesiol Scand,1993,37:105-109
28 Nakae Y,Kanaya N,Namiki A.The direct effects of diazepam and midazolam on myocardial depression in cultured rat ventricular myocytes. Anesth Analg,1997,85:729-733
29 Tsujiguchi N,Yamakage M,Namiki A.Mechanisms of direct inhibitory action of propofol on uterine smooth muscle contraction in pregnant rats [J] . Anesthesiology,2001,95(5):1245-1255
30 Nakae Y,Fujita S,Namiki A.Propofol inhibits Ca2 + transients but not contraction in intact beating guinea pig hearts[J].Anesth Anal ,2000 , 90(6): 1286-1292
31 Sprung J,Ogletree Hughes ML,McConnell BK,et al.The effects of propofol on the contractility of failing and nonfailing human heart muscles[J].Anesth Analg ,2001,93(3):550 - 559
32曹江北,李云峰,米卫东.丙泊酚对细胞内钙离子的调节。国外医学药学分册,2002,29(5):293-296
33 Sato T,Matsuki A,Elemer K,etal. Ketamine relaxes airway smooth muscle contracted by endothelin. Anesth Analg.1997,84:900-906
34聂宏光,崔湧，韩冬云等.氯胺酮对哮喘大鼠气管平滑肌细胞ATP敏感钾电流的影响。中国药理学通报,2003,19(12):1381-1384
35 Robert HB,Elizabeth MW.Mechanisms of broncho protection by anesthetic induction agents.Anesthesiology,1999,90:822-828
36 Christina M,Keith A,Kathleen S,et al.Calcium concentration-dependent mechanisms,through which ketamine relaxes canine airway smooth muscle. Anesthesiology,1997,86:1104-1111
37 Hirota K,Sato T,Rabito SF,et al.Relaxant effect of ketamine and its isomers on histamine-induced contraction oftracheal smooth muscle. Br[J].Anaesth,1996,76:266-270
38周钦海,傅诚章,林桂芳.硫喷妥钠、异丙嗪、氯胺酮、氟哌利多、芬太尼对电压依赖性钙通道的影响.南京医科大学学报,1999,19:496-497
39周钦海,傅诚章,王宁等.蛋白激酶C激动剂PMA、阻滞剂H-7及氯胺酮、咪唑安定对兔离体平滑肌收缩力的影响.南京医科大学学报,2002,22:125-126
40万小健,周钦海,傅诚章等.氯胺酮舒张气道平滑肌作用机制的研究进展。临床麻醉学杂志,2002,18(12):679-680
1 ParmarM,SansomeA.Propofol-induced bronchodilation in status asthmaticus? Anaesthesia.1995 ,50(11):1003-4
2 Eames WO,Rooke GA,Wu RS,etal.Comparison of the effects of etomidate, propofol,and thiopental on respiratory resistance after tracheal intubation. Anesthesiology.1996,84(6):1307-11
3 Wu RS,Wu KC,Sum DC,etal.Comparative effects of thiopentone and propofol on respiratory resistance after tracheal intubation.Br [J]Anaesth. 1996, 77(6):735-8
4 Pizov R,Brown RH,Weiss YS,et al.Wheezing during induction of general anesthesia in patients with and without asthma.A randomized,blinded trial. Anesthesiology.1995,82(5):1111-6
5 Mehr EH,Lindeman KS.Effects of halothane,propofol,and thiopental on peripheral airway reactivity.Anesthesiology.1993,79(2):290-8
6 Cheng EY,Mazzeo AJ,Bosnjak ZJ,et al.Direct relaxant effects of intravenous anesthetics on airway smooth muscle.Anesth Analg. 1996, 83(1):162-8
7 Ouedraogo N,Roux E,Forestier F,et al.Effects of intravenous anesthetics on normal and passively sensitized human isolated airway smooth muscle. Anesthesiology.1998,88(2):317-26
8 Sato T,Matsuki A,Elemer K,etal.Ketamine relaxes airway smooth muscle contracted by endothelin.Anesth Analg.1997,84:900-906
9崔勇,王俊科,孙艳红等.异丙酚、氯胺酮对哮喘大鼠气道痉挛及气道渗出的作用[J].中华麻醉学杂志,2003,23(3):185-187
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11 Sato T,Hirota K.The relaxant effect of ketamine onguinea pig airway smooth muscle is epithelium independent [J].Anesth Analg, 1997, 84: 641-647
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13 Daskalopoulos R,Korcok J,Farhangkhgoee P,etal.Propofol protection of sodium-hydrogen exchange activity sustains glutamate uptake during oxidative stresth[J].Anesth Analg,2001,93:1199-1204
14胡春旭,高崇荣.亚麻醉剂量氯胺酮用于治疗急性严重哮喘9例报道[J].临床麻醉学杂志,2000,16(15):1100-1106
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16 Lau TT,Zed PJ.Does ketamine have a role in managing severe exacerbation of asthma in adults ?[J].Pharmacotherapy,2001,21:1100-1106
17 Szabadkai G, Rizzuto R.Participation of endoplasmic reticulum and mitochondrial calcium handling in apoptosis: more than just neighborhood [J]. FEBSLett,2004,567(1):111-115
18 Verkhratsky A,Toescu EC. Endoplasm in reticulum Ca2+homeostasis and neuronal death [J].J Cell Mol Med,2003,7(4):351-361
19 Bar do S,Cavazzini MG,and Emptage N.The role of the endoplasmic reticulum Ca2+store in the plasticity of central neurons[J].Trends Pharmacology Sci,2006,27(2):78-84
20卢中举,唐大椿,向继洲.平滑肌收缩调节的信号传导.生理科学进展,1997,28:337-340
21孙大业,郭艳林,马力耕.细胞信号转导.生理科学进展,1998.52-168
22 Susan K,Fellen,Willam J.Voltage-gated Ca2+entry and ryanodine receptor Ca2+-induced Ca2+release in preglomerular arterioles.America Physiology, 2007,290(5)
23 Yamakage M, Kohro S, Kawamata T, et al. Inhibitory effects of four inhaled anesthetics on canine tracheal smooth muscle contraction and intracelluar Ca2+ concentration. Anesth Analg, 1993, 77: 67-72
24 Brown RH,Greenberg RS, Wagner EM. Efficiency of propofol to prevent bronchial constriction:effects of preservative. Anesthesiology.2001,94(5):851-5
25周钦海,傅诚章,林桂芳.硫喷妥钠、异丙嗪、氯胺酮、氟哌利多、芬太尼对电压依赖性钙通道的影响.南京医科大学学报,1999,19:496-497
26 Todorovic SM,Perez2 Reyes E,Lingle CJ.Anticonvulsants but not general anesthetics have differential blocking effects on different T2type current variants [J].Mol Pharmacology,2000,58(1):98-108
27 Samain E,Bouillier H,Marty J,etal.The effect of propofol on angiotensin Ⅱ2induced Ca2+ mobilization in aortic smooth muscle cells from normotensive and hypertensive rats [J].Anesth Analg,2000,90(3):546-552
28 Tsutsumi S,Tomioka A , Sudo M, etal . Propofol activates vanilloid receptor channels expressed in human embryonic kidney 293 cells [J].Neurosci Lott ,2001,312(1):45-49
29 Puttick RM, Terror DA. Effects of propofol and effleurage on action potentials membrane currents and contraction of guinea-pig isolated ventricular myocytes[J].Br J Pharma-col,1992,107(2):559 - 565
30 Tsujiguchi N,Yamakage M,Namiki A.Mechanisms of direct inhibitrary action of propofol on uterine smooth muscle contraction in pregnant rats [J].Anesthesiology,2001,95(5):1245-1255
31 Nakae Y,Fujita S,Namiki A. Propofol inhibits Ca2 + transients but not contraction in intact beating guinea pig hearts[J] .Anesth Analg,2000,90 (6):1286-1292
32 Sprung J,Ogletree-Hughes ML, McConnell BK,etal.The effects of propofol on the contractility of failing and nonfailing human heart muscles[J] . Anesth Analg , 2001 , 93(3) :550 - 559
33 Hirota K,Sato T,Rabito SF,etal.Relaxant effect of ketamine and its isomers on histamine-induced contraction of tracheal smooth muscle[J].Br Anaesth, 1996,76:266-270
34王春爱,李红芳,张景华.丙泊酚对离体和在体气管平滑肌收缩活动的抑制作用。卫生职业教育,2007,25(20):120-122
35 Okanlami OA,Fryer AD,Hirshman.Interaction of nondepolarizing muscle relaxants with M2and M3muscarinic receptors in guinea pig lung andheart.Anesthesiology,1996,84:155-161
36 MotohikoH,keithAJ,DavidOW.Effects of intravenous anesthetics on Ca2+ sensitivity in canine tracheal smooth muscle.Anesthesiology,2000,92:133- 139
37 Gordienko DV,Bolton TB.Crosstalk between ryanodine receptors and IP3 receptors as a factor shaping spontaneous Ca2+-release events in rabbit portal vein myocytes.J Physiol 2002;542(3):743-762
38 Horibe M,Kondo I,Damron DS,et al.Propofol attenuates capacitative calcium entry in pulmonary artery smooth muscle cells[J].Anesthesiology, 2001,95(3):681-688
39刘宝刚,曾帮雄.氯胺酮对主肺动脉平滑肌细胞膜钙通道的影响。中华麻醉学杂志,1997,17(7):427
40 Yamakage M,Hirshman CA,Croxton TL.Inhibitory effects of thiopental, ketamine,and propofol on voltage-dependent Ca2+channels in porcine tracheal muscle cells.Anesthesiology,1995,83:1274-1281
41聂宏光,崔湧，韩冬云等.氯胺酮对哮喘大鼠气管平滑肌细胞ATP敏感钾电流的影响。中国药理学通报,2003,19(12):1381-1384
42 Robert HB,Elizabeth MW.Mechanisms of broncho protection by anesthetic induction agents.Anesthesiology,1999,90:822-828
43 Christina M,Keith A,Kathleen S,et al.Calcium concentration-dependent mechanisms,through which ketamine relaxes canine airway smooth muscle. Anesthesiology,1997,86:1104-1111
44 Hirota K,Sato T,Rabito SF,et al.Relaxant effect of ketamine and its isomers on histamine-induced contraction oftracheal smooth muscle.Br J Anaesth, 1996,76:266-270
45周钦海,傅诚章,王宁等.蛋白激酶C激动剂PMA、阻滞剂H-7及氯胺酮、咪唑安定对兔离体平滑肌收缩力的影响.南京医科大学学报,2002,22:125-126
46万小健,周钦海,傅诚章等.氯胺酮舒张气道平滑肌作用机制的研究进展。临床麻醉学杂志,2002,18(12):679-680
2孙大业,郭艳林,马力耕.细胞信号转导.生理科学进展,1998.52-168
3 Pan BX, Zhao GL,Huang XL,Zhao KS.Calcium mobilization is required for peroxynitrite-mediated enhancement of spontaneous transient outward currents in arteriolar smooth muscle cells. Free Radic Biol Med 2004,37(6): 823-838
4 Fellner SK,Parker LA.Endothelin B receptor Ca2+ signaling in shark vascular smooth muscle: participation of inositol trisphosphate and ryanodine receptors.J Exp Biol,2004;207(19):3411-3417
5 Ouedraogo N,Roux E,Forestier F,et al.Effects of intravenous anesthetics on normal and passively sensitized human isolated airway smooth muscle. Anesthesiology.1998,88(2):317-26
6刘宝刚,曾帮雄.氯胺酮对主肺动脉平滑肌细胞膜钙通道的影响。中华麻醉学杂志,1997,17(7):427
7 Susan K,Fellen,Willam J.Voltage-gated Ca2+ entry and ryanodine receptor Ca2+-induced Ca2+ release in preglomerular arterioles.America Physiology, 2007,290(5)
8 Yamakage M,Hirshman CA,Croxton TL.Inhibitory effects of thiopental, ketamine,and propofol on voltage-dependent Ca2+channels in porcine tracheal muscle cells.Anesthesiology,1995,83:1274-1281
9聂宏光,崔湧，韩冬云.氯胺酮对哮喘大鼠气管平滑肌细胞ATP敏感钾电流的影响。中国药理学通报,2003,19(12):1381-1384
10 Sinner B,Friedrich O,Zink W,et al.Ketamine stereoselectively inhibits spontaneous Ca2+-oscillations in cultured hippocampal neurons.Anesth Analg, 2005,100:1660-1666
11 Hardingham GE,Chawla S,Johnson CM, et al. Distinct functions of nuclear and cytoplasmic calcium in the control of gene expression.Nature, 1997,385:260-265
12肖欣荣,陈文彬,程德云.慢性缺氧抑制肺动脉平滑肌细胞钾通道活性[J].中华结核和呼吸杂志,1998,21(11):681-5
13刘杰,赵克森,金春华.重症休克时细动脉平滑肌细胞膜ATP敏感钾通道的变化[J].中华创伤杂志,1998,14(增刊1):10-12
14 Szabadkai G,Rizzuto R. Participation of endoplasmic reticulum and mitochondrial calcium handling in apoptosis:more than just neighborhood [J].FEBSLett,2004,567(1):111-115
15 Verkhratsky A,Toescu EC.Endoplasm in reticulum Ca2+ homeostasis and neuronal death[J].J Cell Mol Med,2003,7(4):351-361
16 Bardo S,Cavazzini MG, Emptage N.The role of the endoplasmic reticulum Ca2+store in the plasticity of central neurons[J].Trends Pharmacol Sci, 2006,27(2):78-84
17卢中举,唐大椿,向继洲.平滑肌收缩调节的信号传导.生理科学进展,1997,28:337-340
18孙大业,郭艳林,马力耕.细胞信号转导.生理科学进展,1998.52-168
19 Missiaen L,Callewaert G,De Smedt H,Parys JB.2-Aminoethoxydiphenyl borate affects the inositol 1,4,5-trisphosphate receptor, the intracellular Ca2+ pump and the non-specific Ca2+ leak from the non-mitochondrial Ca2+ stores in permeabilized cells.Cell Calcium 2001;29(2):111-116
20 Gordienko DV,Bolton TB.Crosstalk between ryanodine receptors and IP3 receptors as a factor shaping spontaneous Ca2+-release events in rabbit portal vein myocytes.J Physiol 2002;542(3): 743-762
21 Susan K,Fellen,Willam J. Voltage-gated Ca2+ entry and ryanodine receptor Ca2+-induced Ca2+ release in preglomerular arterioles.America Physiology, 2007,290(5)
22 Yamakage M, Kohro S, Kawamata T, et al. Inhibitory effects of four inhaled anesthetics on canine tracheal smooth muscle contraction and intracelluar Ca2+ concentration.Anesth Analg,1993,77:67-72
23 liN,YinL,SuZL.Laser Scanning Confocol Microtechnique[M].Beijing: Renmin Military Medical Publishing House,1997,61-64
24魏敏杰,李智,王俊科等.异丙酚舒张血管平滑肌的作用机制.中华麻醉学杂志,1997,17:676-679
25 Cheng EY,Mazzeo AJ,Bosniak ZJ etal.Direct relaxant effects of intravenous anesthetics on airway smooth muscle.Anesth Analg, 1996, 83:162- 168
26 Mehr EH,Lindeman KS.Effects of halothane,propofol,and thiopental on peripheral airway reactivity.Anesthesiology,1993,79:290-298
27 Conti G,Dell’Utri D,Uilardi V,etal.Propofol induces bronchodilation in mechanically ventilated chronic obstructive pulmonary disease(COPD) patients.Acta Anesthesiol Scand,1993,37:105-109
28 Nakae Y,Kanaya N,Namiki A.The direct effects of diazepam and midazolam on myocardial depression in cultured rat ventricular myocytes. Anesth Analg,1997,85:729-733
29 Tsujiguchi N,Yamakage M,Namiki A.Mechanisms of direct inhibitory action of propofol on uterine smooth muscle contraction in pregnant rats [J] . Anesthesiology,2001,95(5):1245-1255
30 Nakae Y,Fujita S,Namiki A.Propofol inhibits Ca2 + transients but not contraction in intact beating guinea pig hearts[J].Anesth Anal ,2000 , 90(6): 1286-1292
31 Sprung J,Ogletree Hughes ML,McConnell BK,et al.The effects of propofol on the contractility of failing and nonfailing human heart muscles[J].Anesth Analg ,2001,93(3):550 - 559
32曹江北,李云峰,米卫东.丙泊酚对细胞内钙离子的调节。国外医学药学分册,2002,29(5):293-296
33 Sato T,Matsuki A,Elemer K,etal. Ketamine relaxes airway smooth muscle contracted by endothelin. Anesth Analg.1997,84:900-906
34聂宏光,崔湧，韩冬云等.氯胺酮对哮喘大鼠气管平滑肌细胞ATP敏感钾电流的影响。中国药理学通报,2003,19(12):1381-1384
35 Robert HB,Elizabeth MW.Mechanisms of broncho protection by anesthetic induction agents.Anesthesiology,1999,90:822-828
36 Christina M,Keith A,Kathleen S,et al.Calcium concentration-dependent mechanisms,through which ketamine relaxes canine airway smooth muscle. Anesthesiology,1997,86:1104-1111
37 Hirota K,Sato T,Rabito SF,et al.Relaxant effect of ketamine and its isomers on histamine-induced contraction oftracheal smooth muscle. Br[J].Anaesth,1996,76:266-270
38周钦海,傅诚章,林桂芳.硫喷妥钠、异丙嗪、氯胺酮、氟哌利多、芬太尼对电压依赖性钙通道的影响.南京医科大学学报,1999,19:496-497
39周钦海,傅诚章,王宁等.蛋白激酶C激动剂PMA、阻滞剂H-7及氯胺酮、咪唑安定对兔离体平滑肌收缩力的影响.南京医科大学学报,2002,22:125-126
40万小健,周钦海,傅诚章等.氯胺酮舒张气道平滑肌作用机制的研究进展。临床麻醉学杂志,2002,18(12):679-680
1 ParmarM,SansomeA.Propofol-induced bronchodilation in status asthmaticus? Anaesthesia.1995 ,50(11):1003-4
2 Eames WO,Rooke GA,Wu RS,etal.Comparison of the effects of etomidate, propofol,and thiopental on respiratory resistance after tracheal intubation. Anesthesiology.1996,84(6):1307-11
3 Wu RS,Wu KC,Sum DC,etal.Comparative effects of thiopentone and propofol on respiratory resistance after tracheal intubation.Br [J]Anaesth. 1996, 77(6):735-8
4 Pizov R,Brown RH,Weiss YS,et al.Wheezing during induction of general anesthesia in patients with and without asthma.A randomized,blinded trial. Anesthesiology.1995,82(5):1111-6
5 Mehr EH,Lindeman KS.Effects of halothane,propofol,and thiopental on peripheral airway reactivity.Anesthesiology.1993,79(2):290-8
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