摘要
目的观察助阳通便膏对便秘模型小鼠结肠组织VIP-CAMP-PKA-AQP3信号通路的影响。方法将作为实验对象的96只雄性BALB/C小鼠随机分成4组,即正常对照组、模型对照组、助阳通便膏组以及麻仁软胶囊组,24只/组。造模予以复方地芬诺酯片混悬液灌胃的方法进行。在模型建立后,每组小鼠对应给予蒸馏水、助阳通便膏及麻仁软胶囊连续行2周灌胃,结束后取材,应用不同方法——RT-PCR法、Wester Blot法及免疫荧光双标法,观测各组小鼠的结肠组织中血管活性肠肽(VIP)、环磷酸腺苷(CAMP)、蛋白激酶(PKA)和水通道蛋白3(AQP3)的表达。结果通过RT-PCR的方法发现,助阳通便膏组VIP、AQP3mRNA表达水平显著高于模型对照组(P<0.05)。通过Wester Blot的方法发现,助阳通便膏组VIP、CAMP、PKA、AQP3蛋白相对表达水平灰度值均显著高于模型对照组(P<0.05),且助阳通便膏组AQP3、CAMP蛋白相对表达水平灰度值较麻仁软胶囊组显著增高(P<0.05),与正常对照组比较,差异无统计学意义(P>0.05)。通过免疫荧光双标的方法发现,助阳通便膏组VIP-AQP3、CAMP-PKA免疫荧光双标表达双阳性细胞数均显著高于模型对照组(P<0.05)。且助阳通便膏组VIP-AQP3双阳性细胞数与正常对照组比较,差异无统计学意义(P>0.05)。结论助阳通便膏治疗慢传输型便秘的作用机制可能是通过提高结肠组织中VIP-CAMP-PKA-AQP3通路表达水平来实现的。
Objective To observe the effect of Zhuyang Tongbian Cream(助阳通便膏)on VIP-CAMP-PKA-AQP3 signaling pathway in colonic tissue of constipation model mice.Methods Ninety-six male BALB/c mice were randomly divided into 4 groups:normal control group,model control group,Zhuyang Tongbian Cream group and Maren Soft Capsule(麻仁软胶囊)group,with 24 mice in each group.The model was established by intragastric administration of the suspension of Compound Diphenoxylate Tablets.After the establishment of the model,the mice in each group were given the distilled water,Zhuyang Tongbian Cream and Maren Soft Capsule by gavage for two weeks.After the experiment,the samples were collected by RT-PCR and Western blot The expressions of VIP,CAMP,PKA and AQP3 in the colonic tissue of mice in each group were observed by using the methods of Western blot and immunofluorescence double labeling.Results The mRNA expression levels of VIP and AQP3 in the Zhuyang Tongbian Cream group were significantly higher than those in the model control group by RT-PCR(P<0.05).Western blot showed that the gray value of VIP,CAMP,PKA and AQP3 protein in Zhuyang Tongbian Cream group was significantly higher than that in model control group(P<0.05),and the gray value of AQP3 and camp protein in Zhuyang Tongbian Cream group was significantly higher than that in Maren Soft Capsule group(P<0.05)and there was no significant difference between Zhuyang Tongbian Cream group and normal control group(P>0.05).The number of double positive cells of VIP-AQP3 and cAMP-PKA in the Zhuyang Tongbian Cream group was significantly higher than that in the model control group(P<0.05).There was no significant difference in the number of VIP-AQP3 double positive cells between the Zhuyang Tongbian Cream group and the normal control group(P>0.05).Conclusion The mechanism of Zhuyang Tongbian Cream in the treatment of slow transit constipation may be realized by increasing the expression levels of VIP-CAMP-PKA-AQP3 pathway in the colon tissue.
作者
王璐
隋楠
WANG Lu;SUI Nan(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China;The Third Afflicted Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110003,Liaoning,China)
出处
《中华中医药学刊》
CAS
北大核心
2022年第5期147-151,277-278,共7页
Chinese Archives of Traditional Chinese Medicine
基金
国家自然科学基金青年科学基金(81503576)
国家重点研发计划(2018YFC1704100,2018YFC1704105)
国家中医药管理局全国中医药创新骨干人才培训项目(国中医药办人教函[2019]91号)
辽宁省便秘病重点实验室项目(辽科发[2016]26号)
作者简介
王璐(1995-),女(满族),辽宁本溪人,硕士研究生,研究方向:中医外科学肛肠;通讯作者:隋楠(1978-),女,辽宁大连人,主任医师,硕士研究生导师,博士后,研究方向:中西医结合治疗肛肠疾病的临床与科研。E-mail:suinan2226@163.com。
