硫化氢对大鼠结肠平滑肌细胞的舒张作用及机制的研究

发布时间：2018-05-10 09:31

本文选题：硫化氢 + 平滑肌细胞　；参考：《天津医科大学》2008年硕士论文

【摘要】： 目的:本课题旨在研究硫化氢(H_2S)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H_2S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H_2S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H_2S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~(-3)mol/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~(-3)mol/L NaSH组的细胞平均长度大于1×10~(-5)mol/L NaSH组(P＜0.05);而1×10~(-4)和1×10~(-3)mol/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~(-6)mol/L Gli,NaSH+1×10~(-5)mol/L Gli,NaSH+1×10~(-4)mol/L Gli和NaSH+1×10~(-3)mol/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~(-6)mol/L至1×10~(-4)mol/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~(-4)mol/L Gli和NaSH+1×10~(-3)mol/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~(-4)mol/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~(-4)mol/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H_2S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H_2S舒张细胞的作用机制之一,由此推测H_2S可能作为一种气体信号分子参与调节胃肠动力作用。
[Abstract]:Objective: to study the effect of H2S) on the contraction and relaxation of isolated colon smooth muscle cells (SMC). Furthermore, whether the effect of H2S on colon smooth muscle cells was induced by changing the activity of ATP sensitive potassium ion channel and K _ S ~ + -ATPase channel was studied, so as to explore the mechanism of H2S regulating gastrointestinal motility. Methods: single colon smooth muscle cell suspension of Wister rats was prepared. Colonic smooth muscle cells of rats were treated with pienadipine, a channel opener of glibenclamide, a channel inhibitor, NaSHH / NaSH Kapk, a H2S donor, and the length of the long axis of the cells was measured by an image analysis system under inverted phase contrast microscope, and the length of the long axis was measured by image analysis system, which was used to measure the length of the long axis of the colonic smooth muscle cells. To observe the effect of drugs on cell contraction and contraction. Results: The average cell length of 1.NaSH group was longer than that of 30120s group (P < 0.05), and the cell relaxation value was 20.48% (P < 0.05), which was larger than that of control group (P < 0.05). Therefore, 60 s was used as the drug reaction time in the later experiment. The average cell length in the 2. 1 脳 10 ~ (-5) and 1 脳 10~(-3)mol/L NaSH groups was larger than that in the control group (P < 0.05). The average cell length in 1 脳 10 ~ (-4) and 1 脳 10~(-3)mol/L NaSH groups was larger than that in 1 脳 10~(-5)mol/L NaSH group (P < 0.05), but there was no significant difference between 1 脳 10 ~ (-4) and 1 脳 10~(-3)mol/L NaSH group (P > 0.05). The average cell length in 3.NaSH 1 脳 10~(-6)mol/L Glix1 脳 10~(-5)mol/L Glix1 脳 10~(-4)mol/L Gli and NaSH 1 脳 10~(-3)mol/L Gli 4 groups was smaller than that in NaSH PBS group (P < 0.05) and larger than that in PBS PBS group (control group P < 0.05). The concentration of Gli in the range of 1 脳 10~(-6)mol/L to 1 脳 10~(-4)mol/L was negatively correlated with the average cell length, that is, with the increase of Gli concentration, the average cell length decreased gradually (P < 0.05), but there was no significant difference between the NaSH 1 脳 10~(-4)mol/L Gli group and NaSH 1 脳 10~(-3)mol/L Gli group (P > 0.05). The average cell length of 4. 1 脳 10 ~ (10) ~ (-6) 10~(-4)mol/L Pin group was significantly higher than that of the control group (P < 0. 01), and the average cell length increased with the increase of Pin concentration. Conclusion: 1.NaSH can produce cell relaxation. When the time of action was 60 s, the relaxation effect of NaSH cells was the most obvious, and the relaxation effect of NaSH cells was in a dose-dependent manner, that is, the relaxation effect of NaSH increased with the increase of concentration in a certain dose range. 2.Gli could attenuate the relaxation of NaSH(1 脳 10 ~ (-4) mol / L in a dose-dependent manner, but Gli could not completely block the relaxation of NaSH. 3.Pin has the same effect of cell relaxation as NaSH. 4. This study suggested that H2S could relax the isolated colon smooth muscle cells of Wister rats, and the activation of K _ S _ (ATP) channel might be one of the mechanisms of H _ (th) _ 2S relaxation. It is suggested that H _ 2S may be a gas signaling molecule involved in regulating gastrointestinal motility.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R363;R574

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