阴虚津亏模型小鼠颌下腺AQP5及cAMP/PKA-CREB信号通路的表达与意义  

Expression and Significance of AQP5 and cAMP/PKA-CREB Signaling Pathway in Submandibular Gland of Mice with Yin-Deficiency and Fluid Consumption

作　　者：崔荣兴 战丽彬[2] 孙晓霞 翁泽斌[1] 陈宁[1] CUI Rongxing;ZHAN Libin;SUN Xiaoxia;WENG Zebin;CHEN Ning(Nanjing University of Chinese Medicine,Nanjing 210023,Jiangsu,China;Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China)

机构地区：[1]南京中医药大学,江苏南京210023 [2]辽宁中医药大学,辽宁沈阳110847

出　　处：《中华中医药学刊》2022年第6期149-153,I0042,共6页Chinese Archives of Traditional Chinese Medicine

基　　金：国家重点研发计划(2018YFC1704400,2018YFC1704405);国家中医药领军人才岐黄学者支持项目(国家中医药人教函〔2022〕6);江苏省研究生科研与实践创新计划(KYCX20_1547)。

摘　　要：目的观察阴虚津亏模型小鼠颌下腺AQP5、cAMP/PKA-CREB信号通路分子表达变化,探讨阴虚口干发生的生物学基础。方法SPF级C57BLKS/J雄性小鼠随机分为正常组、模型组,使用“热盛伤阴”法建立模型,连续8周。观察小鼠肛温与“五心”温度、唾液流率、粪便含水量、皮肤含水量、尿胆红素、饮食量、体质量变化情况。造模结束后,旷场实验观察小鼠行为变化情况,HE染色观察颌下腺组织病理变化,免疫组化观察AQP5分布表达,Western Blot观察AQP5、p-PKA/PKA、p-CREB/CREB表达。结果与正常组比较,模型组小鼠饮水量显著增多(P<0.0001),唾液流率显著降低(P<0.05),摄食量显著减少(P<0.0001);心前区、左上肢爪心、左下肢爪心以及右下肢爪心温度显著升高(P<0.05,P<0.01);尿胆红素阳性6例,阳性率66.7%;体质量、肛温、右上肢爪心温度、粪便含水量、皮肤含水量差异均无统计学意义(P>0.05)。与正常组比较,模型组小鼠中央格穿格次数与停留时间百分比差异不显著,旷区运动距离增加(P<0.01),运动速度提高(P<0.001);模型组小鼠颌下腺出现浆液性腺泡细胞萎缩、分泌管与微血管扩张等病理变化,颌下腺AQP5免疫组化表达显著低于正常组(P<0.01);模型组小鼠颌下腺AQP5、p-PKA/PKA、p-CREB/CREB的表达水平较正常组均明显降低(P<0.05,P<0.001)。结论阴虚口干的发生与颌下腺组织病变有关,且颌下腺AQP5表达量降低可能是阴虚口干的重要诱因,该变化可能是由cAMP/PKA-CREB信号通路调控。Objective To observe the changes in the molecular expressions of aquaporin 5(AQP5)and cAMP/PKA-CREB signaling pathway in the submandibular glands of mice with Yin-deficiency and fluid consumption to explore the biological basis of the occurrence of dry-mouth symptom of Yin deficiency.Methods SPF-grade C57BLKS/J male mice were randomly divided into the normal group and the model group.The model was established by the method of“excessive heat injuring Yin”for 8 weeks.The changes of mouse rectal temperature and the“temperature of the chest,palms and soles”,saliva flow rate,fecal water content,skin water content,urine bilirubin,diet and body weight were observed.After modeling,the behavioral changes of mice were observed by the open field experiment.The pathological changes of submandibular glands were observed by HE staining.And the distribution and expression of AQP5 were observed by immunohistochemistry.The expressions of AQP5,p-PKA/PKA and p-CREB/CREB were observed by Western Blot.Results Compared with that of the normal group,the mice in the model group significantly drank more water(P<0.0001).The saliva flow rate was significantly reduced(P<0.05)and the food in-take was significantly reduced(P<0.0001).The temperature of the anterior heart area,left upper limb paw center,left lower limb paw center and right lower limb paw center was increased significantly(P<0.05,P<0.01,P<0.01,P<0.01).The urine bilirubin was positive in 6 cases,with a positive rate of 66.7%.There were no statistically significant differences in body mass,rectal temperature,right upper limb paw temperature,fecal water content or skin water content.Compared with those of the nor-mal group,there was no significant difference of the number of central grid crossings or the percentage of residence time in the model group.The distance of movement in the open field was increased(P<0.01),and the movement speed was increased(P<0.001).Pathological changes such as serous acinar cell atrophy,secretion ducts and telangiectasia in the submandibular gla

关 键 词：阴虚津亏 颌下腺 AQP5 cAMP/PKA-CREB信号通路 

分 类 号：R332[医药卫生—人体生理学]