四君子汤对脾虚大鼠腮腺唾液淀粉酶分泌障碍及VIP-cAMP信号通路的影响

目的:观察四君子汤对脾虚大鼠唾液淀粉酶分泌障碍治疗效果,并揭示其作用机制。方法:大鼠皮下注射利血平0.5 mg/kg,正常对照组大鼠注射等量生理盐水,共8 d;造模结束后模型动物分为模型组和四君子汤组,四君子汤组灌胃给予四君子汤,模型组和正常对照组灌胃给予等量双蒸水,给药4 w后进行取材。麻醉动物,取左侧腮腺,缝合伤口,大鼠稍清醒时,用10%冰乙酸溶液20μL刺激大鼠舌尖,1次/min,共刺激30次;处死,取下右侧腮腺。检测腮腺组织唾液淀粉酶活性(sAA)c、AMP及VIP含量、PKA活性和VAMP8、SNAP-23蛋白表达。结果:四君子汤组大鼠腮腺内sAA与正常对照组大鼠无显著差异,模型组大鼠酸刺激后sAA活性显著高于正常对照组大鼠,酸刺激前VIP含量、PKA活性各组大鼠无显著差异;酸刺激后,正常对照组大鼠VIP及cAMP含量、PKA活性显著升高,模型组大鼠VIP轻微升高,cAMP含量、PKA活性显著低于正常对照组,四君子汤组大鼠VIP及cAMP含量、PKA活性有一定程度恢复。模型组VAMP-8蛋白表达与正常对照组比较无显著差异,SNAP-23蛋白表达有所下降;与模型组比较,四君子汤组大鼠SNAP-23、VAMP-8蛋白表达增强。结论:四君子汤对利血平脾虚大鼠唾液淀粉酶分泌障碍有一定疗效,可能通过恢复脾虚大鼠腮腺细胞VIP-cAMP信号通路改变起到治疗作用,包括提高VIP含量和提高SNAP-23和VAMP-8蛋白表达。

Effect of sijunzi decoction on salivary amylase secretion disorder and VIP-cAMP signaling pathway in splenasthenic rats

Objective:To observe the effect of Sijunzi Decoction on secretion disorder of salivary amylase in splenasthenic rat and its mechanism.Methods:The model group rats received reserpine 0.5 mg/kg through subcutaneous injection while the control group rats received the same volume of saline for 8 days.After being modeled,the model group were divided into treatment group and model control group,treatment group were given orally Sijunzi Decoction,model control group and normal group were fed the same amount of distilled water for 4 weeks.The animal were anaesthetized and the left parotid was removed,the wounds were sutured.When the animals were awake but drowsy,20μL 10% glacial acetic acid was applied on the apex of the tongue once a minute for 30 minutes,removed the right parotid gland of the animals.The samples were frozen and amylase activity and VIP,cyclic adenosine monophosphate(cAMP)content and VAMP-8、SNAP-23 protein expression in the parotid glands were detected.Results:Change of sAA in parotid acinar was not significantly different between treatment group and normal groups,but higher in model control groups after acid stimulation.The VIP and PKA contents were not significantly different among three groups.VIP,cAMP content and PKA activity increased significantly in normal group while VIP increased slightly,cAMP and PKA activity decreased in model control groups,which returned to some degrees in treatment group after acid stimulation.Expression of VAMP-8 protein was not significantly different between treatment group and model control groups,while expression of SNAP-23 was lower in model control groups,expression of VAMP-8 and SNAP-23 was higher in treatment group than which in model control groups.Conclusion:Sijunzi Decoction has a certain effect on secretion disorder of salivary amylase in splenasthenic rat,which mechanism may be related to recover changes of VIP-cAMP signal pathway in the splenasthenic rat′s parotid gland cells,including increase VIP content and expression of VAMP-8 and SNAP-23.