基于AKT-HK2-NF-κB信号通路的辣椒素对CIA大鼠的作用机制  

Mechanism of capsaicin in CIA rats based on AKT-HK2-NF-κB signaling pathway

作　　者：郑云松 皮浩吕 何子晗 雷欢欢 马书怡 吴宁 吴遵秋 ZHENG Yunsong;PI Haolyu;HE Zihan;LEI Huanhuan;MA Shuyi;WU Ning;WU Zunqiu(Laboratory of Chemisty and Biochemistry,School of Basic Medicine,Guizhou Medical University,Guiyang 550525,Guizhou,China)

机构地区：[1]贵州医科大学基础医学院化学与生物化学实验室,贵州贵阳550525

出　　处：《贵州医科大学学报》2024年第11期1615-1621,共7页Journal of Guizhou Medical University

基　　金：国家自然科学基金(82260829);贵州省中药管理局项目(QZYY-2022-014);2021年省级大学生创新创业计划训练项目(S202110660034)。

摘　　要：目的探讨辣椒素(CAP)对大鼠类风湿性关节炎(RA)蛋白质丝氨酸苏氨酸激酶(AKT)/己糖激酶2(HK2)/核转录因子-κB(NF-κB)信号通路的作用及机制。方法SD大鼠尾根部皮下注入牛Ⅱ型胶原与弗氏完全佐剂的乳化剂,构建胶原诱导型关节炎(CIA)大鼠模型作为模型组,空白组大鼠尾根部注入等体积的生理盐水;将造模成功的SD大鼠随机分为CAP低、中、高剂量组(CAP剂量分别为1、2、3 mg/kg)以及阳性对照组(雷公藤剂量为40 mg/kg),于造模成功2周后灌胃3周,空白组与模型组灌胃等量生理盐水;造模前后及给药期间测量大鼠足肿胀程度,给药3周取大鼠踝关节切片、HE染色观察组织学变化,Western blot检测大鼠关节滑膜组织AKT、HK2、NF-κB的表达,RT-qPCR检测AKT、HK2、NF-κb mRNA的表达,ELISE检测大鼠血清中白细胞介素-6(IL-6)含量。结果与模型组相比,CAP高剂量组大鼠右后足AI评分降低(P<0.05),大鼠足肿胀度减轻(P<0.05);与阳性对照组相比,CAP高剂量组大鼠右后足AI评分及大鼠足肿胀度差异无统计学意义(P>0.05);HE染色显示,与空白组相比,模型组大鼠踝关节区滑膜出现增生,软骨下骨质结构破坏,同时伴有炎性细胞的浸润和滑膜血管翳的侵袭;与模型组相比,阳性对照组及CAP各组大鼠在进行药物干预后,关节滑膜组织增生均得到不同程度的抑制,且CAP高、中剂量组大鼠关节滑膜组织内浸润的炎症细胞以及新生血管数均有所减少;与模型组比较,阳性对照组和CAP高剂量组IL-6含量降低(P<0.05);与模型组大鼠比较,CAP高、中剂量组及阳性对照组大鼠滑膜组织AKT、HK2、NF-κB mRNA及蛋白表达水平下降(P<0.05)。结论高剂量CAP可减轻RA大鼠踝关节的炎症反应,其机制可能与抑制AKT-HK2-NF-κB通路相关蛋白及降低血清IL-6水平有关。Objective To explore the role of capsaicin(CAP)on the protein-serine-threonine kinase(Akt)/hexokinase 2(HK2)/nuclear factor kappa-B(NF-κB)signaling pathway in rats with rheumatoid arthritis(RA)and its mechanism.Methods Emulsion of bovine typeⅡcollagen with complete Freund's adjuvant(CFA)was subcutaneously inject into the tail roots of SD rats to construct a collagen-induced arthritis(CIA)rat model as model group.An equal volume of physiological saline was injected into rat tail roots in blank group.The successfully modeled SD rats were randomly divided into low-dose CAP group(1 mg/kg),medium-dose CAP group(2 mg/kg),high-dose CAP group(3 mg/kg)and positive control group(Tripterygium wilfordii,40 mg/kg).At two weeks after successful modeling,blank and model groups were given an equal amount of physiological saline by gavage.The degree of rat foot swelling was measured before modeling,after modeling and during drug administration.After 3 weeks of administration,ankle joints were sectioned and stained with HE.Western blot was used to detect the expression of AKT,HK2,and NF-κB in rat synovial tissues.RT-qPCR was applied to detect the mRNA expressions of AKT,HK2,and NF-κB.ELISA was applied to detect the content of rat serum interleukin-6(IL-6).Results When compared to model group,high-dose CAP group showed a decrease in AI score in right hind feet of the rats(P<0.05)and a reduction in foot swelling(P<0.05).There were no statistically significant differences in AI score of the right hind foot and the degree of foot swelling between positive control group and high-dose CAP group(P>0.05).HE staining revealed that when compared to blank group,rat ankle synovial membrane in model group had significant proliferation,destruction of subchondral bone structure,accompanied by inflammatory cell infiltration and invasion of synovial membrane blood vessels.When compared to model group,the rats in positive control and CAP groups showed varying degrees of inhibition of synovial tissue proliferation after drug intervention,and in

关 键 词：类风湿性关节炎 辣椒素 蛋白激酶B 核因子-κB 己糖激酶2 白细胞介素-6 

分 类 号：R34[医药卫生—基础医学]