| Objective:To observe and analyze the relationship between the mechanism of Jinxueyuan Granules increased the saliva secretion of xerostomia model rats and excitement of the receptors,providing an experimental basis for further study of the signaling pathways which Jinxueyuan Granules effectiveness of nourishing Yin and generating body fluid involved,and providing an experimental basis for the traditional Chinese medicine theory"Body fluid and blood from the same source".Methods:By using M3-receptor blocker 4-Diphenyl-acetoxy-N-methyl-piperidine(4-DAMP),M-receptor blocker atropine,and adrenergic receptor blocker phentolamine intravenously,three groups of xerostomia model rats were successfully established.The saline group was the negative control group,with M1 receptor agonist pilocarpine group as the positive control group.After 20 minutes of establishing acute model rats,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 After administration,the dry cotton ball with 30 milligram weight was placed in the sublingual of rats to measure the salivary secretion.Each cotton ball was changed every 30 minutes,and measured 150 minutes continuously.The maximum amount of saliva secretion was calculated through measuring weight difference of dry cotton ball before and after the study in order to observe the effects of Jinxueyuan Granules in saliva secretion inhibited by each receptor blocker.After 30 minutes of repeatedly administration,submandibular gland tissue was isolated,then the effect of Jinxueyuan Granules for expression of AQP5 in submandibular gland cells was analyzed by the Western blot technology,the AQP5 protein expression level of each group was compared with a density of Image Hyper ? image analysis software.Results:1.The secretion of saliva in rats were inhibited severely by atropine,the saliva secretion was significantly reduced,compared with normal control group(P<0.05).In the saliva secretion of model group induced by saline and Jinxueyuan Granules,Jinxueyuan Granules could significantly induce the saliva secretion of the model rats of xerostomia(P<0.05).There was no significant difference between the low-dose of Jinxueyuan Granules group and the saline group(P>0.05),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).In the recently 50 minutes after administration of the Jinxueyuan Granules,there was an obvious increasing in the amount of saliva secretion,and then decreased rapidly,with a slow growth trend,higher than the control group.2.The saliva secretion of rats were subjected to different degrees of inhibition caused by 4-DAMP and atropine,the saliva secretion was significantly reduced,compared with the normal group(P<0.05),with atropine group obviously.And gavaged Jinxueyuan Granules on the basis of two blockers,it was found that the saliva secretion of the drug groups were increased,compared with the normal group and the model groups,there was a statistical significance(P<0.05).3.Adrenergic receptor blocker phentolamine mildly inhibited the saliva secretion of rats.Compared with the normal group,the saliva secretion was reduced(P<0.05).And 20 minutes later,gavaged Jinxueyuan Granules.It was found that the saliva secretion of rats had been restored and increased,compared with the model group(P<0.05).4.AQP5 protein was expressed in the submandibular glands of rats in the normol control group;compared with normol group,acetylcholine agonists pilocarpine could obviously induce and increase the protein expression of AQP5.After the administration of three blockers,the expression of AQP5 protein in submandibular gland cells were decreased evidently,compared with the normal group(P<0.05).After administration of Jinxueyuan Granules,the AQP5 expression levels in blockers groups had been restored and increased,compared with the model groups significantly(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(P>0.05).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).Conclusion:Jinxueyuan Granules can restore the saliva secretion and the AQP5 protein expression inhibited by the adrenal receptor blocker and M3 receptor blocker,and increase the saliva secretion of rats.These results suggest 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.But which specific components in Jinxueyuan Granules associated with these receptors,and the pathway mediated by receptors need further study. |
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