摘要
目的探究杨梅素(myricetin,Myr)调节环磷酸腺苷(cyclic adenosine monophosphate,cAMP)/蛋白激酶A(protein kinaseA,PKA)/cAMP反应成分结合蛋白(cAMP-responseelementbindingprotein,CREB)信号通路对炎症性肠病(inflammatory bowel disease,IBD)大鼠免疫功能的影响。方法建立IBD大鼠模型,实验分为对照组、模型组、Myr低、中、高剂量(Myr-L、Myr-M、Myr-H,28、56、112 mg·kg^(-1)·d^(-1)Myr)组和Myr高剂量+PKA抑制剂H89(MyrH+H89,112 mg·kg^(-1)·d^(-1)Myr+7 mg·kg^(-1)·d^(-1)H89)组。对大鼠疾病活动指数(DAI)进行评分;测定免疫功能指标及结肠长度;试剂盒测定血清中IL-6、IL-17A、TNF-α、cAMP水平;HE染色观察结肠组织病理变化;流式细胞术测定Treg细胞比例;免疫组化检测结肠组织中MPO表达;Western blotting测定cAMP/PKA/CREB信号通路相关蛋白。结果与对照组相比,模型组结肠组织细胞排列紊乱、有大量炎性细胞浸润,出现严重溃疡现象,大量细胞坏死,黏膜水肿,DAI评分、IL-6、TNF-α和IL-17A水平、脾脏系数、胸腺系数、MPO光密度值显著增加(P<0.05),结肠长度、Treg细胞比例、cAMP浓度、p-PKA/PKA和p-CREB/CREB水平显著降低(P<0.05)。与模型组相比,Myr-L、Myr-M和Myr-H组结肠组织细胞排列较整齐,黏膜水肿、炎性细胞浸润、细胞坏死及溃疡现象减少,DAI评分、IL-6、TNF-α和IL-17A水平、脾脏系数、胸腺系数、MPO光密度值显著降低(P<0.05),结肠长度、Treg细胞比例、cAMP浓度、p-PKA/PKA和pCREB/CREB水平逐渐增加(P<0.05)。与Myr-H组相比,Myr-H+H89组结肠组织病理变化加重,DAI评分、IL-6、TNF-α和IL-17A水平、脾脏系数、胸腺系数、MPO光密度值显著增加(P<0.05),结肠长度、Treg细胞比例、cAMP浓度、pPKA/PKA和p-CREB/CREB水平显著降低(P<0.05)。结论Myr可能通过激活cAMP/PKA/CREB信号通路抑制机体炎症水平,调节免疫功能,对IBD大鼠发挥保护作用。
OBJECTIVE To investigate the effect of myricetin(Myr)on immune function in rats with inflammatory bowel disease(IBD)by regulating the cAMP/PKA/CREB signaling pathway.METHODS IBD rat models were established and separated into control group,model group,low,medium,and high dose Myr(Myr-L,Myr-M,Myr-H,28,56,112 mg·kg^(-1)·d^(-1)Myr)groups,and high dose Myr+PKA inhibitor H89(Myr-H+H89112 mg·kg^(-1)·d^(-1)Myr+7 mg·kg^(-1)·d^(-1)H89)group.The disease activity index(DAI)of rats was scored;immune function indicators and colon length were measured;the levels of IL-6,IL-17A,TNF-α,and cAMP in serum were determined by the kit;the pathological changes of colon tissue were observed by HE staining;the proportion of Treg cells was determined by flow cytometry;immunohistochemistry was used to detect the expression of MPO in colon tissue;Western blotting was used to determine cAMP/PKA/CREB signaling pathway related proteins.RESULTS Compared with the control group,the colon tissue cells in the model group were disorderly arranged,with a large number of inflammatory cell infiltration,severe ulceration,a large number of cell necrosis,mucosal edema,the DAI score,IL-6,TNF-α,and IL-17A levels,spleen coefficient,thymus coefficient,and MPO optical density values were obviously increased(P<0.05),the colon length,Treg cell ratio,cAMP concentration,p-PKA/PKA,and p-CREB/CREB levels were obviously reduced(P<0.05).Compared with the model group,the arrangement of colon tissue cells in the Myr-L,Myr-M,and Myr-H groups was relatively neat;mucosal edema inflammatory cell infiltration,cell necrosis and ulcer phenomenon were reduced;the DAI score,IL-6,TNF-α,and IL-17A levels,spleen coefficient,thymus coefficient,and MPO optical density values were gradually reduced(P<0.05);the colon length,Treg cell ratio,cAMP concentration,p-PKA/PKA,and p-CREB/CREB levels were gradually increased(P<0.05).Compared with the Myr-H group,the pathological changes in the colon tissue of the Myr-H+H89 group worsened,the DAI score,IL-6,TNF-α,and IL-17A levels,spleen
作者
周亚妮
李若楠
ZHOU Yani;LI Ruonan(College of Health Management,Shangluo University,Shangluo 726000,China;Shangluo Health Industry Engineering Technology Research Center,Shangluo 726000,China)
出处
《中国现代应用药学》
CAS
CSCD
北大核心
2024年第11期1456-1463,共8页
Chinese Journal of Modern Applied Pharmacy
基金
陕西省科技计划项目(2020JM-630)
商洛市科学技术研究发展计划项目(2020-Z-0061)
国家级大学生创新创业训练计划项目(S202311396018)。
关键词
杨梅素
环磷酸腺苷/蛋白激酶A/cAMP反应成分结合蛋白信号通路
炎症性肠病
免疫功能
myricetin
cyclic adenosine monophosphate
protein kinase A
cyclic adenosine monophosphate responsive component binding protein(AMP
PKA
CREB)signaling pathway
inflammatory bowel disease
immune function
