大鼠胃平滑肌细胞收缩模型及其药物研究

目的 建立胃平滑肌收缩的实验模型并且对相应治疗药物的作用进行评价。方法 采用分离大鼠胃底平滑肌,用胶原酶和胰蛋白酶进行消化培养,得到离体平滑肌细胞悬液。用Trypan blue稀释法检查,通过带刻度显微镜观察和标测细胞的形态和长度,并观察在不同的时间范围内细胞形态和长度的变化情况,计算细胞存活率和存活时间。于细胞悬液加入各种神经介质乙酰胆碱(ACh)、5-羟色胺(5-HT、组织胺(His)引起刺激反应,同时加入其相应的拮抗剂阿托品(atropine)、多潘立酮(Domp)、苯海拉明(Diph)时,观察细胞形态和长度的前后变化,从而对模型和药物的作用进行评价。结果 每只大鼠胃底均能收获1×10~6以上个细胞,细胞存活率在95％以上,细胞至少可存活2.5 h。ACh能引起胃平滑肌细胞的收缩,具有明显的量效关系,1×10~(-8)mol·L~(-1)即可产生最大收缩反应(38.0％)。atropine对乙酰胆碱的拮抗作用有明显的量效关系。5-HT对大鼠胃底平滑肌细胞具有明显的收缩作用,井有明显的量效关系。Domp能够拮抗5-HT的细胞收缩作用,并具有一定的量效关系。5-HT的量效曲线比较平坦,可能至少有2种受体参与细胞的收缩作用,它们引起细胞收缩的最大效应分别为20.7％和10.5％。His引起的平滑肌细胞收缩,在1×10~(-13)～1×10~(-10) mol·L~(-1)内有?

The model of stomach smooth muscle cells contraction of rats and the effects of treatment drugs in vitro

AIM To set up the model of stom- effects of its treatment drugs. METHODS To ap-ach smooth muscle cells contraction and assess the ply the culture of isolated stomach fundus smooth muscle cells in rats in vitro and to obtain the mixture of stomach smooth muscle cells. The cells were examined with the Trypan blue dilution technology under microscope and the length of cells and the contraction rate of muscle cells were adopted to the evaluation index of effects. The change of cells in different time was observed and calculating the survival rate and time of cells. The selected neuro-medium and its antagonist, such as acetyl-choline chloride (ACh) and atropine, 5-HT and domperidone(Domp) , histamine( His) and diphen-hydramine(Diph) were added to the mixture cells so as to provoke irritant reaction observe the effects of drugs. RESULTS Obtaining cells in stomach fundus smooth muscle was more than 1 X 106 in each rat by this way, and the survival rate and time of cells was 95% and 2. 5 h. The contraction of cells was observed after adding ACh 5-HT or His ,and the effects of these agents were antag- onized by antagonist atropine, Domp and Diph respectively. The effects of maximum contraction of ACh in concentration 1×10-8 mol · L-1 was 38%, and in 5-HT may exist two receptors, and the maximum rate was 20. 7% and 10. 5% respectively. The effective dose of contraction was 1 X 10-13 -1×10-10 mol · L-1 induced by His, the contraction effects was significantly weakened in high of 1 ×10-10 mol · L-1 and antagonized by Diph. CONCLUSION The smooth muscle cells by digesting stomach fundus smooth muscle of rats would be more than 95% survivors and survival time was more than 2. 5 h. ACh,5-HT and His could induce the contraction of smooth muscle cells, and the effects of contraction were antagonized by atropine, Domp and Diph respectively.