本文選題：硫化氫 + 平滑肌細胞　； 參考：《天津醫(yī)科大學》2008年碩士論文

【摘要】： 目的:本課題旨在研究硫化氫(H_2S)對離體單個結(jié)腸平滑肌細胞舒縮作用的影響;并進一步研究H_2S對結(jié)腸平滑肌細胞的舒縮作用是否通過改變ATP敏感鉀離子通道(K_(ATP)通道)的活性引起,從而探討H_2S調(diào)節(jié)胃腸道動力的機制。 方法:制備Wister大鼠單個結(jié)腸平滑肌細胞懸液。用H_2S供體硫氫化鈉(NaSH),NaSH+K_(ATP)通道抑制劑格列本脲(Gli),K_(ATP)通道開放劑吡那地爾(Pin)處理大鼠結(jié)腸平滑肌細胞,在倒置相差顯微鏡下觀察,應(yīng)用圖象分析系統(tǒng)測量細胞長軸的長度,觀察藥物對細胞舒縮作用的影響。 結(jié)果: 1.NaSH組作用時間為60s組的細胞平均長度大于作用時間為30、120s組(P＜0.05)并大于對照組(P＜0.05),其細胞舒張量值為20.48%。故后期實驗中均選用60s作為藥物反應(yīng)時間。 2.1×10~(-5)、1×10~(-4)和1×10~(-3)mol/L NaSH組的細胞平均長度均大于對照組(P＜0.05)。其中1×10~(-4)和1×10~(-3)mol/L NaSH組的細胞平均長度大于1×10~(-5)mol/L NaSH組(P＜0.05);而1×10~(-4)和1×10~(-3)mol/L NaSH組之間的細胞平均長度無統(tǒng)計學差異(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組的細胞平均長度均小于NaSH+PBS組(P＜0.05),大于PBS+PBS組(對照組)(P＜0.05)。其中Gli在1×10~(-6)mol/L至1×10~(-4)mol/L范圍內(nèi)其濃度與細胞平均長度呈負相關(guān)趨勢,即隨著Gli濃度增加細胞平均長度逐漸減少(P＜0.05);而NaSH+1×10~(-4)mol/L Gli和NaSH+1×10~(-3)mol/L Gli組之間的細胞平均長度無統(tǒng)計學差異(P＞0.05)。 4.1×10~(-6)、1×10~(-5)、1×10~(-4)mol/L Pin組的細胞平均長度較對照組增加(P＜0.01),且隨著Pin濃度的增加,細胞平均長度增加。 結(jié)論: 1.NaSH可以產(chǎn)生細胞舒張作用。在作用時間為60s時,其舒張作用最明顯;且NaSH的細胞舒張作用具有一定的劑量依賴性,即在一定劑量范圍內(nèi)其舒張作用隨濃度的增加而增強。 2.Gli可以減弱NaSH(1×10~(-4)mol/L)對細胞的舒張作用,并具有一定的劑量依賴性;但Gli不能完全阻斷NaSH的細胞舒張作用。 3.Pin可以和NaSH一樣產(chǎn)生細胞舒張作用。 4.本研究提示H_2S可以對Wister大鼠離體單個結(jié)腸平滑肌細胞產(chǎn)生舒張作用,激活K_(ATP)通道可能是H_2S舒張細胞的作用機制之一,由此推測H_2S可能作為一種氣體信號分子參與調(diào)節(jié)胃腸動力作用。
[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.
【學位授予單位】：天津醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2008
【分類號】：R363;R574

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