清泻浊毒法对类风湿关节炎大鼠作用靶点及免疫调节机制分析
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摘要
目的:分析清泻浊毒法对类风湿关节炎(RA)大鼠作用靶点及免疫调节机制。方法:选取SPF级Wistar雄性大鼠45只,随机数字表法将大鼠分成三组,模型组、正常组与观察组,各15只,模型组与观察组大鼠制备RA模型,成功造模后观察组大鼠灌胃0. 5ml/100g的清泻浊毒方药液,模型组和对照组大鼠灌胃等剂量生理盐水,每天一次,连续灌胃21天;观察大鼠活动、饮食及毛色等一般状况,造模前及给药后7天、14天及21天时称量各组大鼠体重,是用数显游标卡尺与特制细皮尺对大鼠足垫厚度与踝关节周长进行测量,ELISA法检测血清白细胞介素-1 (IL-1)、IL-6及IL-10含量,免疫检测大鼠关节组织内TLR2、TLR3及TLR4表达状况。结果:给药7天、14天、12天后模型组大鼠体重比正常组下降,观察组大鼠体重比模型组上升,模型组及观察组大鼠踝关节周径、踝足垫厚度比正常上升,差异均有统计学意义(P<0. 05);模型组大鼠血清IL-1、IL-6含量比正常组上升,IL-10含量比正常组下降,观察组大鼠血清IL-1、IL-6含量比模型组下降,IL-10含量比模型组上升,差异均有统计学意义(P<0. 05);模型组大鼠关节组织内TLR2、TLR3及TLR4表达量比正常组上升,观察组比模型组下降,差异均有统计学意义(P<0. 05)。结论:清泻浊毒法可影响RA大鼠体内TLRs信号传导路径,对炎症因子表达起到调节作用,控制大鼠关节内炎症发展。
Objective: To analyze the target and immune regulation mechanism of Qingxie Zhuodu Method for rheumatoid arthritis( RA) rats. Methods: A total of 45 Wistar male rats of SPF grade were randomly divided into three groups: model group,normal group and observation group,15 rats in each group. RA model of the model group and the observation group was successfully prepared. Rats in the observation group were intragastrically administered with 0. 5 ml/100 g of Qingxie Zhuodu Recipe solution. The rats in the model group and the controlled group were intragastrically administered with normal saline once a day for 21 days. The rats were observed for activities,diet and in general conditions such as coat color,the body weight of each group was weighed before modeling and at 7,14 and 21 days after administration. The thickness of the footpad and the circumference of the ankle joint were measured by a digital vernier caliper and a special fine tape measure. The levels of serum interleukin-1( IL-1),IL-6 and IL-10 were detected by immunoassay. The expressions of TLR2,TLR3 and TLR4 in rat joint tissues were detected by immunoassay. Results: After 7,14,and 12 days of administration,the body weight of the model group was lower than that of the normal group. The weight of the rats in the observation group was higher than that of the model group. The thickness of the ankle joint circumference and the thickness of the footpad of the model group and the observation group increased normally. The differences were statistically significant( P < 0. 05). The levels of serum IL-1 and IL-6 in the model group were higher than those in the normal group,and the IL-10 content was lower than that in the normal group. The serum IL-1 and IL-6 in the observation group were observed. The content of 6 decreased compared with the model group,and the IL-10 content increased compared with the model group,the difference was statistically significant( P < 0. 05). The expression of TLR2,TLR3 and TLR4 in the joint tissue of the model group was higher than that of the normal group. The model group decreased,and the difference was statistically significant( P < 0. 05). Conclusion: Qingxie Zhuodu Method can affect the signal transduction pathway of TLRs in RA rats,regulate the expression of inflammatory factors,and control the development of inflammation in rat joints.
Objective: To analyze the target and immune regulation mechanism of Qingxie Zhuodu Method for rheumatoid arthritis( RA) rats. Methods: A total of 45 Wistar male rats of SPF grade were randomly divided into three groups: model group,normal group and observation group,15 rats in each group. RA model of the model group and the observation group was successfully prepared. Rats in the observation group were intragastrically administered with 0. 5 ml/100 g of Qingxie Zhuodu Recipe solution. The rats in the model group and the controlled group were intragastrically administered with normal saline once a day for 21 days. The rats were observed for activities,diet and in general conditions such as coat color,the body weight of each group was weighed before modeling and at 7,14 and 21 days after administration. The thickness of the footpad and the circumference of the ankle joint were measured by a digital vernier caliper and a special fine tape measure. The levels of serum interleukin-1( IL-1),IL-6 and IL-10 were detected by immunoassay. The expressions of TLR2,TLR3 and TLR4 in rat joint tissues were detected by immunoassay. Results: After 7,14,and 12 days of administration,the body weight of the model group was lower than that of the normal group. The weight of the rats in the observation group was higher than that of the model group. The thickness of the ankle joint circumference and the thickness of the footpad of the model group and the observation group increased normally. The differences were statistically significant( P < 0. 05). The levels of serum IL-1 and IL-6 in the model group were higher than those in the normal group,and the IL-10 content was lower than that in the normal group. The serum IL-1 and IL-6 in the observation group were observed. The content of 6 decreased compared with the model group,and the IL-10 content increased compared with the model group,the difference was statistically significant( P < 0. 05). The expression of TLR2,TLR3 and TLR4 in the joint tissue of the model group was higher than that of the normal group. The model group decreased,and the difference was statistically significant( P < 0. 05). Conclusion: Qingxie Zhuodu Method can affect the signal transduction pathway of TLRs in RA rats,regulate the expression of inflammatory factors,and control the development of inflammation in rat joints.
引文
[1]姚文琴,王定瑜,刘元忠,等.狭叶崖爬藤对类风湿关节炎大鼠的免疫调节作用[J].医药导报,2016,35(2):141~143
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[11]张引红,李美宁,王春芳,等.补骨脂素对类风湿性关节炎小鼠模型的免疫调节作用[J].中国实验动物学报,2017,25(2):207~210
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[13]张瑶.祛风舒筋丸对佐剂关节炎大鼠免疫调节及VEGF、ICAM-1表达的影响[J].安徽医药,2017,18(4):653~655
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[15]王凤杰,樊莎莎,陈显兵,等. Lunasin对实验性类风湿关节炎大鼠滑膜成纤维细胞增殖与凋亡的影响[J].免疫学杂志,2016,4(2):114~118
[16]周文旭,方堃,谭湘淑,等.甲氨蝶呤对类风湿性关节炎大鼠滑膜细胞凋亡基因调控的实验研究[J].陕西医学杂志,2018,7(4):105~108
[2]梁卫,梁涛,张丽玲,等.风湿痹痛方治疗大鼠胶原性关节炎的实验研究[J].中国中医急症,2017,26(1):83~85
[3] Jia Z,Jiao H E. Paeoniflorin ameliorates rheumatoid arthritis inrat models through oxidative stress, inflammation and cyclooxygenase 2[J]. Experimental Therapeutic Medicine,2016,11(2):655~659
[4]童萍,何东仪.雷公藤及其提取物对类风湿关节炎免疫调节机制的研究进展[J].现代免疫学,2016,5(3):250~252
[5]彭代平,汤小虎,周瑞彬.桂枝温通经脉配伍对胶原诱导性关节炎大鼠的免疫调节作用[J].中国中西医结合杂志,2017,37(7):813~818
[6] Doss H M,Ganesan R,Rasool M. Trikatu,an herbal com-pound ameliorates rheumatoid arthritis by the suppression of inflammatoryimmune responses in rats with adjuvant-induced arthritis and on cultured fi-broblast like synoviocytes via the inhibition of the NFκB signaling pathway.[J]. Chemico-Biological Interactions,2016,25(19):175~186
[7]陈玉兴,卓俊城,黄雪君,等.痹痛消胶囊对大鼠类风湿性关节炎的作用及其机制[J].中成药,2016,38(6):1211~1215
[8]吴雪莲,吕洪华.风湿祛痛汤对实验性关节炎大鼠的治疗作用及机制研究[J].中药材,2017,13(7):1722~1726
[9] Abhari K,Shekarforoush S S,Hosseinzadeh S, et al. Theeffects of orally administeredBacillus coagulansand inulin on prevention andprogression of rheumatoid arthritis in rats[J]. Food Nutrition Research,2016,60(39):1~8
[10]胡华刚,陶黎,吴佳莹,等.藏药二十五味驴血丸对痹症(类风湿关节炎)大鼠的治疗作用及其机理探讨[J].中国中医基础医学杂志,2016,14(8):1053~1056
[11]张引红,李美宁,王春芳,等.补骨脂素对类风湿性关节炎小鼠模型的免疫调节作用[J].中国实验动物学报,2017,25(2):207~210
[12]刘彩丽,王颖瑛. TNF-α/NF-κB信号通路在依那西普缓解大鼠类风湿性关节炎中的作用[J].中国应用生理学杂志,2017,33(4):373~376
[13]张瑶.祛风舒筋丸对佐剂关节炎大鼠免疫调节及VEGF、ICAM-1表达的影响[J].安徽医药,2017,18(4):653~655
[14]高远,李永吉.类风湿性关节炎发病机制及中药靶向治疗的研究进展[J].中国药房,2016,27(35):5030~5034
[15]王凤杰,樊莎莎,陈显兵,等. Lunasin对实验性类风湿关节炎大鼠滑膜成纤维细胞增殖与凋亡的影响[J].免疫学杂志,2016,4(2):114~118
[16]周文旭,方堃,谭湘淑,等.甲氨蝶呤对类风湿性关节炎大鼠滑膜细胞凋亡基因调控的实验研究[J].陕西医学杂志,2018,7(4):105~108