摘要
目的:研究当归拈痛汤对风湿热痹型佐剂性关节炎大鼠的防治作用及对自噬蛋白微管相关蛋白1轻链3(LC3),自噬关键分子酵母Atg6同系物(Beclin1),p62表达的影响。方法:将6周龄SD雄性大鼠随机分为正常组,风湿热痹模型组,当归拈痛汤低、中、高剂量组(5.67,11.34,22.68 g·kg^(-1)),甲氨蝶呤(MTX)组(1.35 mg·kg^(-1)),每组10只,通过对大鼠尾根部皮下注射灭活结核分支杆菌佐剂建立佐剂性关节炎大鼠模型,用人工气候箱进行风湿热痹证模型造模16 d,于免疫当天灌胃给药,持续28 d。进行大鼠一般情况观察及足趾肿胀度、关节炎指数(AI)检测,苏木素-伊红(HE)染色观察大鼠膝关节滑膜组织病理形态学改变,实时荧光定量聚合酶链式反应(Real-time PCR)检测大鼠滑膜组织LC3,Beclin1,p62的mRNA表达,蛋白免疫印迹法(Western blot)及免疫组化法分别检测大鼠滑膜组织LC3,Beclin1,p62的蛋白表达。结果:与正常组比较,风湿热痹模型组大鼠给药16,20,24,28 d时足趾肿胀度显著升高(P<0.01),AI显著升高(P<0.01),滑膜细胞增生显著,自噬蛋白LC3,Beclin1蛋白及mRNA表达显著上调(P<0.01),p62蛋白及mRNA表达显著下调(P<0.01);与风湿热痹模型组比较,各给药组大鼠足趾肿胀度减轻,滑膜增生情况有不同程度改善,LC3,Beclin1的mRNA和蛋白表达水平显著降低(P<0.01),p62的mRNA和蛋白表达水平明显升高(P<0.05,P<0.01),且当归拈痛汤中剂量组防治效果最佳。结论:当归拈痛汤对风湿热痹型佐剂性关节炎大鼠有明显的防治作用,且这种防治作用可能与调控滑膜组织中自噬相关因子LC3,Beclin1,p62的表达,抑制自噬有关。
Objective: To observe the preventive and control effects of Danggui Niantongtang against adjuvant arthritis differentiated into wind-damp-heat impediment in rats and its influences on the expression of autophagy-related proteins microtubule-associated protein 1 light chain 3(LC3), homolog of yeast Atg6(Beclin1)and p62. Method: The six-week-old male SD rats were randomly divided into the normal group,wind-damp-heat impediment model group,low-,medium-,and high-dose Danggui Niantongtang(5.67,11.34,22.68 g·kg^(-1))groups,and methotrexate(MTX,1.35 mg·kg^(-1))group,with 10 rats in each group. A rat model of adjuvant arthritis was established by subcutaneous injection of inactivated Mycobacterium tuberculosis into the tail root,followed by exposure to the manual climatic box for 16 d for inducing the wind-damp-heat impediment.The drugs were administered intragastrically on the day of immunization for 28 d. The general conditions of rats were observed and the swelling degree of toes and arthritis index(AI)were detected. The pathological changes in the synovial tissues of the knee joints were observed by hematoxylin-eosin(HE)staining. The mRNA expression levels of LC3,Beclin1,and p62 in the synovial tissues were measured by real-time fluorescence quantitative polymerase chain reaction(Real-time PCR),followed by the assay of their protein expression by Western blot and immunohistochemistry. Result: Compared with the normal group, the wind-damp-heat impediment model group exhibited significantly increased swelling degree of toes(P<0.01),increased AI(P<0.01),proliferated synovial cells(P<0.01),up-regulated LC3 and Beclin1 protein and mRNA expression(P<0.01),and down-regulated p62 protein and mRNA expression(P<0.01)after 16,20,24,28-d medication.Compared with the wind-damp-heat impediment model group,each medication group displayed alleviated toe swelling and synovial hyperplasia to different degrees,decreased mRNA and protein expression levels of LC3 and Beclin1(P<0.01),and increased p62 mRNA and protein expression(P<0.05,
作者
陆麒瑾
李佳钰
蔡义思
赵帆
林鸿鑫
刘刚
袁立霞
LU Qi-jin;LI Jia-yu;CAI Yi-si;ZHAO Fan;LIN Hong-xin;LIU Gang;YUAN Li-xia(School of Traditional Chinese Medicine,Southern Medical University,Guangzhou 510515,China;Shenzhen Hospital of Guangzhou University of Chinese Medicine(Futian),Shenzhen 518034,China;Nanfang Hospital of Southern Medical University,Guangzhou 510515,China;The Fifth Affiliated Hospital of Southern Medical University,Guangzhou 5\0920,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2022年第1期41-49,共9页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81673875,81774382)
广东省中医药局课题项目(20181175)。
