参榆洗液改善痔术后大鼠痛觉敏化及对cAMP/PKA信号通路的影响  

Effect of Shenyu Lotion(参榆洗液)on Pain Sensitization and cAMP/PKA Signaling Pathway in Rats after Hemorrhoid Surgery

作　　者：李惠雯 严建[2] 宾东华[2] 胡婷 LI Huiwen;YAN Jian;BIN Donghua;HU Ting(Hunan University of Chinese Medicine,Changsha Hunan 410208,China;The First Hospital of Hunan University of Chinese Medicine,Changsha Hunan 410007,China)

机构地区：[1]湖南中医药大学,湖南长沙410208 [2]湖南中医药大学第一附属医院,湖南长沙410007

出　　处：《中医药导报》2024年第10期41-47,共7页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：湖南省中医药科研计划项目(B2023035)。

摘　　要：目的:基于环磷酸腺苷/蛋白激酶A(c AMP/PKA)信号通路探究参榆洗液对痔术后大鼠疼痛的影响。方法:构建痔术后大鼠模型,将造模成功的56只SD大鼠随机分为模型组、溶剂模型组、参榆洗液组、参榆洗液+Forskolin(c AMP/PKA信号通路激动剂)组、Forskolin组、H-89(c AMP/PKA信号通路抑制剂)组、NS-398(PGE_(2)抑制剂)组,每组8只;另取8只大鼠为空白组。各组大鼠给予对应药物干预,连续7 d。给药结束后1 d,观察大鼠一般状态及疼痛相关动物学行为,观察创面变化并计算创面愈合率,苏木精-伊红(HE)染色观察各组大鼠创面组织病理学改变,蛋白质免疫印迹法(Western blotting)检测各组大鼠背根神经节TRPV1、p-PKA蛋白表达及创面组织中c AMP、PKA蛋白表达,实时荧光定量聚合酶链式反应(Real-time PCR)检测大鼠创面组织中c AMP m RNA、PKA m RNA表达,酶联免疫吸附测定法(ELISA)检测创面组织中PGE_(2)、IL-6、IL-1β含量。结果:模型组大鼠背根神经节组织中p-PKA、TRPV1及创面周围组织c AMP、PKA、PGE_(2)、c AMP m RNA、PKA m RNA、IL-6、IL-1β表达水平升高(P<0.05);与模型组比较,H-89组、NS-398组、参榆洗液+Forskolin组、参榆洗液组创面组织病变程度减轻,创面愈合率依次上升,背根神经节组织中p-PKA、TRPV1及创面组织c AMP、PKA、c AMP m RNA、PKA m RNA、IL-6、PGE_(2)、IL-1β表达水平依次降低(P<0.05);Forskolin干预能显著提高痔术后大鼠模型c AMP、PKA、TRPV1、p-PKA、PGE_(2)、IL-6、IL-1β表达水平(P<0.05),与参榆洗液联合干预后,上述蛋白及炎症因子表达水平显著下降(P<0.05)。结论:参榆洗液对痔术后大鼠cAMP/PKA信号通路有抑制作用,抑制c AMP/PKA信号通路可改善痔术后痛觉敏化,其机制可能与抑制PGE_(2)表达、减少炎症因子释放、抑制TRPV1活化有关。Objective:To investigate the effect of Shenyu lotion on postoperative hemorrhoidal pain in rats based on cyclic adenosine monophosphate-protein kinase A(cAMP-PKA)signaling pathway.MethodS:A rat model of postoperative hemorrhoidal pain was constructed,and 56 SD rats were randomly divided into model group,solvent model group,Shenyu lotion group,Shenyu lotion+Forskolin(cAMP-PKA signaling pathway agonist)group,Forskolin group,H-89(cAMP-PKA signaling pathway inhibitor)group,NS-398(PGE_(2)inhibitor)group,8 rats in each group.Another 8 rats was selected as blank group.The rats in each group were given the corresponding drug intervention for 7 consecutive days.One day after the drug administration,the general state and pain-related zoological behaviors of the rats were observed.The changes in the wounds were observed and the wound healing rate was calculated.Histopathological changes in the wounds of the rats were observed by hematoxylin-eosin(HE)staining in each group.The protein expression of TRPV1 and p-PKA in the dorsal root ganglion and the protein expression of cAMP and PKA in the wounds were detected by Western blotting.Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)was used to detect the expression levels of cAMP mRNA and PKA mRNA in the traumatized tissues of rats.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of PGE_(2),IL-6 and IL-1βin the traumatized tissues.Results:The expression levels of p-PKA and TRPV1 in dorsal root ganglion tissues,and cAMP,PKA,PGE_(2),cAMP mRNA,PKA mRNA,IL-6 and IL-1βin periwound tissues increased in model group(P<0.05).Compared with the model group,the degree of wound tissue lesions was reduced,and the wound healing rate increased sequentially in H-89 group,NS-398 group,Shenyu lotion+Forskolin group,and Shenyu lotion group.The expression levels of p-PKA,TRPV1 in the dorsal root ganglion tissue,as well as cAMP,PKA,cAMP mRNA,PKA mRNA,IL-6,PGE_(2),and IL-1βin the wound tissue,decreased sequentially in H-89 group,NS-398 group,and Shen

关 键 词：痔术后疼痛 参榆洗液 cAMP/PKA信号通路 PGE_(2) TRPV1 大鼠 

分 类 号：R285.5[医药卫生—中药学]