津血源增加口干症模型鼠唾液分泌机制的实验研究

为分析津血源的具体作用机制，该实验对津血源增加口干症模型SD大鼠唾液分泌与激动受体之间的关系进行了研究。研究通过使用M受体阻滞剂4-二苯乙酰氧基-N-甲基-哌啶（4-DAMP）和阿托品，或肾上腺受体阻滞剂酚妥拉明，制作3组口干症模型大鼠；造模成功后，予中药津血源灌胃，根据临床常用剂量及SD大鼠体表面积换算，阿托品组将实验动物分为津血源低、中、高剂量组，4-DAMP及酚妥拉明组灌胃中剂量津血源。所有动物给药后，连续测定150 min的唾液分泌量，观察津血源增加唾液分泌的作用特点及与受体之间的关系；并分离大鼠的颌下腺组织，通过Western blot分析津血源对颌下腺细胞水分子通道蛋白5（AQP5）表达的影响。结果发现，比较各组唾液分泌量，与生理盐水组、酚妥拉明组、4-DAMP组及阿托品组相比，津血源组的唾液分泌明显增加，并存在明显差异，P<0.05。在阿托品组中，津血源低剂量组与生理盐水组疗效相当，而津血源中剂量组与生理盐水组相比，存在统计学差异（P<0.05）。唾液分泌量增加的时间曲线图中，发现津血源组与生理盐水组之间存在统计学差异（P<0.05）。与各阻断剂组相比，津血源治疗组颌下腺细胞AQP5蛋白量的表达较前增加，P<0.05 ；除阿托品组外，津血源对其余2组阻断剂的疗效与生理盐水组无明显差异。对津血源给药后的3种阻断剂AQP5的表达量进行比较，发现津血源对阿托品组的疗效比4-DAMP、酚妥拉明组更明显（P<0.05），后两者之间并不存在统计学差异。因此，研究认为津血源增加唾液分泌（养阴生津作用）的机制可能是通过毒蕈碱M受体（尤其是M₃受体）、肾上腺素受体介导的作用途径，增加唾液腺细胞膜AQP5的表达，促进唾液分泌。

Research of mechanism Jinxueyuan granules increased saliva secretion of xerostomia model rats

To analyze the specific mechanism of Jinxueyuan granules, the relationship between the Jinxueyuan granules increased the saliva secretion of xerostomia model SD rats and excitement of receptors were studied in this experiment.In the study,three groups of xerostomia model rats were successfully established by using M-receptor blockers-4-diphenyl-acetoxy-N-methyl-piperidine (4-DAMP) and atropine,or adrenergic receptor blocker phentolamine;after the modeling,the medicine Jinxueyuan granules were gavaged.According to the clinical dose of Jinxueyuan granules and SD rats body surface area,the rats in atropine group were divided three dose groups respectively,namely low,medium and high dose of Jinxueyuan granules groups.The 4-DAMP group and phentolamine group were gavaged medium dose of Jinxueyuan granules.And the amount of salivary secretion for 150 minutes in all groups continuously were measured,and the effect of Jinxueyuan granules increased salivation and the relationship between characteristics and the receptors were observed;and submandibular gland tissue of the rats was isolated,then the effect of Jinxueyuan Granules for expression of the water channel protein aquaporin-5 (AQP5) in submandibular gland cells was analyzed by the Western blot technology.It was found that the saliva secretion of Jinxueyuan Granules groups was increased significantly,and compared with the saline control group, phentolamine group,4-DAMP group and atropine group,difference was significant, P<0.05.There was no significant difference between the low-dose of Jinxueyuan granules group and the saline group, but the medium dose of Jinxueyuan granules group had a significant difference,compared with the saline group(P<0.05). In the time distribution of increasing saliva secretion,there was a significant difference between the saline and Jinxueyuan granules group in the saliva secretion (P<0.05).After administration of Jinxueyuan granules, the expression of AQP5 protein in the submandibular gland cells expressing of treatment groups was increased,and compared with the blocker groups, there was a significant difference, P<0.05.Except the atropine group,there was no significant difference in Jinxueyuan granules relieving the inhibition induced by blocks in phentolamine group and 4-DAMP group,compared with the saline group.Compared the AQP5 expression in three blockers groups,there was no significant difference in the efficacy of Jinxueyuan granules between phentolamine group and 4-DAMP group;but there was a significant difference between the atropine gruop and other groups(P<0.05).Therefore,it was considered that the mechanism of Jinxueyuan granules increasing saliva secretion (effectiveness of nourishing Yin and generating body fluid) possibly through the pathway mediated by muscarinic M receptor,especially M₃ receptor,or adrenergic receptor, and increased expression of salivary gland AQP5 membrane, and then stimulate saliva production.