【作者】 程英；

【导师】 王邦茂； 杨玉龙；

【作者基本信息】 天津医科大学 ， 内科学， 2008， 硕士

【摘要】 目的:本课题旨在研究硫化氢（H₂S）对离体单个结肠平滑肌细胞舒缩作用的影响;并进一步研究H₂S对结肠平滑肌细胞的舒缩作用是否通过改变ATP敏感钾离子通道(K_ATP通道)的活性引起,从而探讨H₂S调节胃肠道动力的机制。方法:制备Wister大鼠单个结肠平滑肌细胞悬液。用H₂S供体硫氢化钠（NaSH）,NaSH+K_ATP通道抑制剂格列本脲（Gli）,K_ATP通道开放剂吡那地尔（Pin）处理大鼠结肠平滑肌细胞,在倒置相差显微镜下观察,应用图象分析系统测量细胞长轴的长度,观察药物对细胞舒缩作用的影响。结果:1.NaSH组作用时间为60s组的细胞平均长度大于作用时间为30、120s组（P＜0.05）并大于对照组（P＜0.05）,其细胞舒张量值为20.48%。故后期实验中均选用60s作为药物反应时间。2.1×10^-5、1×10^-4和1×10^-3mol/L NaSH组的细胞平均长度均大于对照组（P＜0.05）。其中1×10^-4和1×10^-3mol/L NaSH组的细胞平均长度大于1×10^-5mol/L NaSH组（P＜0.05）;而1×10^-4和1×10^-3mol/L NaSH组之间的细胞平均长度无统计学差异（P＞0.05）。3.NaSH+1×10^-6mol/L Gli,NaSH+1×10^-5mol/L Gli,NaSH+1×10^-4mol/L Gli和NaSH+1×10^-3mol/L Gli 4组的细胞平均长度均小于NaSH+PBS组（P＜0.05）,大于PBS+PBS组（对照组）（P＜0.05）。其中Gli在1×10^-6mol/L至1×10^-4mol/L范围内其浓度与细胞平均长度呈负相关趋势,即随着Gli浓度增加细胞平均长度逐渐减少（P＜0.05）;而NaSH+1×10^-4mol/L Gli和NaSH+1×10^-3mol/L Gli组之间的细胞平均长度无统计学差异（P＞0.05）。4.1×10^-6、1×10^-5、1×10^-4mol/L Pin组的细胞平均长度较对照组增加（P＜0.01）,且随着Pin浓度的增加,细胞平均长度增加。结论:1.NaSH可以产生细胞舒张作用。在作用时间为60s时,其舒张作用最明显;且NaSH的细胞舒张作用具有一定的剂量依赖性,即在一定剂量范围内其舒张作用随浓度的增加而增强。2.Gli可以减弱NaSH(1×10^-4mol/L)对细胞的舒张作用,并具有一定的剂量依赖性;但Gli不能完全阻断NaSH的细胞舒张作用。3.Pin可以和NaSH一样产生细胞舒张作用。4.本研究提示H₂S可以对Wister大鼠离体单个结肠平滑肌细胞产生舒张作用,激活K_ATP通道可能是H₂S舒张细胞的作用机制之一,由此推测H₂S可能作为一种气体信号分子参与调节胃肠动力作用。更多还原

【Abstract】 Objective:To study the effect of hydrogen sulfide（H₂S）on the size of single colonic smooth muscle cell in vitro and to investigate whether the effect of H₂S on colonic smooth muscle cell size is mediated by ATP-sensitive potassium channel(K_ATP channel).So that to know how H₂S works on gastrointestinal motility and what is its molecule mechanism.Methods:.The single colonic smooth cells suspension of Wister rats were prepared. Sodium hydrosulfide（NaSH）（H₂S donor）,both NaSH and Glibenclamide（Gli） (K_ATPchannel inhibitor),and Pinacidil（Pin）(K_ATPchannel opener)were added in cells suspension respectively.Cells were observed under the inverted phase contrast microscope and the sizes were measured with an image analysis system to evaluate the effects of drugs.Results:1.In NaSH group,the cells were longer at 60 seconds than the cells at 30 or 120 seconds（P＜0.05）and they were also longer than the control group（P＜0.05）, with a relaxation value of 20.48%.So,an action time of 60 seconds was employed in the later experiments.2.The cells added by NaSH at 1×10^-5,1×10^-4and 1×10^-3mol/L were longer than the control group（P＜0.05）.The cells treated with NaSH at 1×10^-3and 1×10^-4mol/L were longer than those treated with NaSH at 1×10^-5mol/L （P＜0.05）.No statistical significance was found in the length of cells between the group treated with NaSH at 1×10^-4mol/L and the group treated with NaSH at 1×10^-3mol/L（P＞0.05）.3.The cells treated with NaSH + 1×10^-6mol/L Gli,NaSH + 1×10^-5mol/L Gli, NaSH + 1×10^-4mol/L Gli,NaSH + 1×10^-3mol/L Gli were all shorter than the cells treated with NaSH +PBS and longer than the cells treated with PBS +PBS（the control group）（P＜0.05）.Negative correlation was found between the Gli concentration and the cell length.As the Gli concentration was increased from 1×10^-6to 1×10^-4mol/L,the average cell length were becoming shorter （P＜0.05）.There was no statistical significance in the cell length between the cells treated with NaSH + 1×10^-4mol/L Gli and those treated with NaSH+ 1×10^-3mol/L Gli（P＞0.05）.4.The cells were longer in Pin group(1×10^-6,1×10^-5and 1×10^-4mol/L respectively)than the control group（P＜0.05）.The cell length increased as the concentration of Pin increased.Conclusion:1.Cells could be relaxed by NaSH.NaSH with an action time of 60 seconds had the largest relaxation effect on the colonic smooth cells.In addition,a dose-dependent relaxation effect could be founed.In a certain dosage range,the relaxation effect enhanced as the concentration of NaSH increased.2.The relaxation effect of cells induced by NaSH could be attenuated by Gli and a dose-dependent inhibition effect could be found.But the relaxation effect of NaSH could not be blocked by Gli completely.3.Pin had the same effect on cells relaxation as NaSH.4.H₂S might produce the relaxation effect on colonic smooth muscle cells of wister rats,which mightbe partly due to the activation of K_ATPchannel.It could be inferred that H₂S was probably involved in the regulation of the gastrointestinal motility as a gaseous signal molecule.更多还原