三七皂苷R1对小鼠颞下颌关节疼痛和肠道菌群的影响  

Effect of notoginsenoside R1 on temporomandibular joint pain and gut flora in mice

作　　者：王沛 王静[1] 何岚[1] 宁琼琼 李汉超[1] WANG Pei;WANG Jing;HE Lan;NING Qiongqiong;LI Hanchao(Department of Rheumatology and Immunology,First Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710016,China)

机构地区：[1]西安交通大学第一附属医院风湿免疫科,西安710016

出　　处：《山西医科大学学报》2025年第1期66-75,共10页Journal of Shanxi Medical University

基　　金：国家自然科学基金面上项目(82371811)。

摘　　要：目的 探讨三七皂苷R1(NGR1)对小鼠颞下颌关节疼痛(TMJP)的缓解作用及机制,分析NGR1对肠道菌群的影响。方法 将C57BL/6J小鼠随机分为5组(n=12):假手术组、TMJP组、12.5NGR1组、25NGR1组和50NGR1组。假手术组小鼠双侧颞下颌关节(TMJ)腔内注射10μL生理盐水,其他组小鼠双侧TMJ腔内注射完全弗氏佐剂(5 mg/mL)诱导TMJP小鼠模型。注射完成后1 h,12.5NGR1组、25NGR1组和50NGR1组小鼠分别腹腔注射12.5,25,50 mg/kg的NGR1,其他组腹腔注射生理盐水,每天注射1次,连续4 d。给药结束后24 h,测定小鼠面部机械刺激疼痛阈值。HE染色观察颞下颌关节软骨组织形态,甲苯胺蓝(TB)染色检测颞下颌关节软骨组织蛋白聚糖水平。免疫组化法检测颞下颌关节软骨组织基质金属蛋白酶13(MMP13)和白细胞介素(IL)-1β的表达。ELISA法检测血清IL-1β、肿瘤坏死因子-α(TNF-α)和IL-6水平。免疫荧光染色检测脑组织Iba-1和胶质纤维酸性蛋白(GFAP)表达。RT-qPCR和Western blot分别检测脑组织中诱导型一氧化氮合成酶(iNOS)、IL-1β、TNF-α、IL-10和精氨酸酶-1(Arg-1)的mRNA和蛋白表达水平。RT-qPCR分析粪便样本中拟杆菌门(Bacteroidetes)和毛螺菌科(Lachnospiraceae)细菌含量。结果 与假手术组相比,TMJP组小鼠的面部机械刺激疼痛阈值降低(P<0.05),颞下颌关节出现明显病变,颞下颌关节软骨组织蛋白聚糖含量降低(P<0.05),MMP13和IL-1β水平升高(P<0.05),血清TNF-α、IL-1β和IL-6水平升高(P<0.05),脑组织中Iba-1相对荧光强度升高(P<0.05),脑组织中iNOS、IL-1β和TNF-α的mRNA和蛋白水平升高(P<0.05),拟杆菌门和毛螺菌科菌群的含量降低(P<0.05)。与TMJP组相比,12.5NGR1组、25NGR1组和50NGR1组小鼠的面部机械刺激疼痛阈值升高(P<0.05),颞下颌关节病变减轻,颞下颌关节软骨组织蛋白聚糖含量升高(P<0.05),MMP13和IL-1β水平降低(P<0.05),血清TNF-α、IL-1β和IL-6水平降低(P<0.05),脑组织中Iba-1相对荧光�Objective To investigate the alleviating effect and mechanism of notoginsenoside R1(NGR1)on temporomandibular joint pain(TMJP)in mice and analyze the effect of NGR1 on gut flora.Methods C57BL/6J mice were randomly divided into 5 groups(n=12):sham group,TMJP group,12.5NGR1 group,25NGR1 group and 50NGR1 group.The mice in sham group were injected with 10μL of normal saline into the bilateral temporomandibular joint(TMJ)cavity.The mice in the other groups were injected with complete Freund's adjuvant(5 mg/mL)into the bilateral TMJ cavity to induce the TMJP mouse model.One hour after the CFA injection,the mice in 12.5NGR1 group,25NGR1 group and 50NGR1 group were intraperitoneally injected with NGR1 at a dose of 12.5,25 and 50 mg/kg,respectively.The mice in the other groups were intraperitoneally injected with normal saline.The mice in each group were in-jected once a day for 4 consecutive days.At 24 h after the end of the administration,the pain threshold of facial mechanical stimulation was measured.HE staining was used to evaluate the morphology of the temporomandibular joint cartilage tissue,and toluidine blue(TB)staining was used to detect the level of proteoglycan in the temporomandibular joint cartilage tissue.The expressions of matrix metalloproteinase 13(MMP13)and interleukin-1β(IL-1β)in temporomandibular joint cartilage were detected by immunohistochemical staining.Serum levels of IL-1β,tumor necrosis factor-α(TNF-α)and IL-6 were detected by enzyme-linked immunosorbent assay(ELISA).The expression of Iba-1 and glial fibrillary acidic protein(GFAP)in brain tissue was detected by immunofluorescence staining.The mRNA and protein expression levels of inducible nitric oxide synthase(iNOS),IL-1β,TNF-α,IL-10 and arginase-1(Arg-1)in brain tissue were analyzed by RT-qPCR and Western blot.The contents of Bacteroidetes and Lachnospiraceae in stool samples were analyzed by RT-qPCR.Results Compared with sham group,the facial mechanical stimulation pain threshold of mice was de-creased in TMJP group(P<0.05),the temp

关 键 词：颞下颌关节 疼痛 三七皂苷R1 小胶质细胞 肠道菌群 表型 

分 类 号：R782[医药卫生—口腔医学]