运脾解毒通络祛湿方对胶原诱导性关节炎大鼠炎症因子表达及脂质代谢的影响

Effects of Yunpi Jiedu Tongluo Qushi Formula on expression of inflammatory factors and lipid metabolism in rats with collagen-induced arthritis

目的:探究运脾解毒通络祛湿方对胶原诱导性关节炎(CIA)大鼠脂质代谢及相关炎症因子的影响,并寻找二者的相关性。方法:32只雌性Wistar大鼠适应性喂养1周后,随机分为对照组、模型组、中药组、西药组,每组8只。以胶原注射的方式构建CIA模型,在第14天分别以单蒸水、运脾解毒通络祛湿方、甲氨蝶呤干预3周,期间每周评价各组大鼠的体质量及双后肢足趾容积,从第14天开始每周评价各组关节炎症评分。ELISA法检测初次免疫后第21、35天大鼠血清中转化生长因子β1(TGF-β1)、肿瘤坏死因子α(TNF-α)、白细胞介素-17A(IL-17A)水平。对初次免疫后第35天大鼠血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)、总蛋白(TP)及肌酐(Crea)含量进行检测。利用多维质谱“鸟枪法”脂质组学技术对磷脂酰肌醇(PI)、神经酰胺(CER)、磷脂酰丝氨酸(PS)、4-羟基壬烯醛(HNE)、溶血磷脂酰胆碱(LPC)、鞘磷脂(SM)、磷脂酰甘油(PG)7大类脂质分子进行分析;对具有差异的脂质分子与炎症因子进行Pearson相关性分析。结果:各组大鼠体质量无明显变化。与模型组比较,各治疗组关节炎症评分显著下降(P<0.05,P<0.01),西药组大鼠足趾肿胀度显著降低(P<0.05),中药组足趾肿胀度也得到了一定的缓解。药物干预后,各治疗组血清IL-17A、TGF-β1、TNF-α水平较模型组均显著下降(P<0.01)。各治疗组ALT、AST、TP含量无显著性差异,而Crea含量显著下降(P<0.01,P<0.05)。药物干预后,与模型组比较,PI含量显著下降(P<0.01)。PI脂质分子与炎症因子的Pearson相关性分析结果显示,PI脂质分子与大鼠血清中IL-17A、TGF-β1、TNF-α水平具有显著的正相关性(P<0.01,P<0.05)。结论:CIA大鼠模型中存在PI类脂质代谢异常,且该脂质代谢异常与炎症相关,运脾解毒通络祛湿方可通过调节PI类脂质的代谢,降低炎症水平,减轻关节肿胀,改善类风湿关节炎的疾病状态,且该方对肝肾功能无显著影响。

Objective:To investigate the effect of Yunpi Jiedu Tongluo Qushi Formula on lipid metabolism and related inflammatory factors in collagen-induced arthritis(CIA)rats,and to find the correlation between the two.Methods:A total of 32 female Wistar rats were randomly divided into control group,model group,traditional Chinese medicine group and western medicine group after adaptive feeding for one week,8 rats each group.The CIA model was constructed by collagen injection.On the 14th day,single distilled water,Yunpi Jiedu Tongluo Qushi Decoction,and methotrexate were used to intervene for 3 weeks.During the period,the body weight and hind limbs of the rats in each group were evaluated every week.Toe volume,and arthritis scores in each group were assessed weekly from day 14.The serum levels of TGF-β1,TNF-αand IL-17A were detected by ELISA on the 21st and 35th days after the primary immunization.The contents of ALT,AST,TP and Crea in the serum of rats were detected on the 35th day after the primary immunization.The MDMS-SL technology was used to analyze 7 major lipid molecules including PI,CER,PS,HNE,LPC,SM,PG;Pearson correlation analysis was performed on lipid molecules with differences and inflammatory factors.Results:The body weight of the rats in each group did not change significantly.Compared with the model group,the arthritis scores in each treatment group were significantly decreased(P<0.05,P<0.01),the degree of toe swelling of the rats in the western medicine group was significantly reduced(P<0.05),and the degree of toe swelling in the traditional Chinese medicine group was also relieved to a certain exten.After drug intervention,the levels of serum IL-17A,TGF-β1 and TNF-αin each treatment group were significantly lower than those in the model group(P<0.01).There was no significant difference in the contents of ALT,AST and TP among the treatment groups,while the Crea content decreased significantly(P<0.01,P<0.05).Through qualitative and quantitative analysis of MDMS-SL,compared with the model group,the contents of PI decreased significantly after the intervention of traditional Chinese medicine and western medicine(P<0.01).The Pearson correlation analysis of PI lipid molecules and inflammatory factors showed that PI lipid molecules were significantly correlated with the levels of IL-17A,TGF-β1,and TNF-αin rat serum,all of which were positively correlated(P<0.01,P<0.05).Conclusion:There is abnormal PI lipid metabolism in CIA rat model,and the abnormal lipid metabolism is related to inflammation.Yunpi Jiedu Tongluo Qushi Formula can regulate the metabolism of PI lipids,reduce the level of inflammation,relieve joint swelling and improve the disease state of RA,and it has no significant effect on liver and kidney function.