中医正骨2025，Vol.37Issue(5)：30-41,12.
独活-秦艽对膝骨关节炎模型大鼠的影响及其作用机制研究
Effects of Duhuo(独活)-Qinjiao(秦艽)on knee osteoarthritis in model rats and its mechanism:an experimen-tal study
摘要
Abstract
Objective:To observe the therapeutic effects of Duhuo(Angelicae Pubescentis Radix(TCD))-Qinjiao(Gentianae Macro-phyllae Radix)(DH-QJ)on knee osteoarthritis(KOA)in model rats,and to explore its underlying mechanism.Methods:Fifty-six male SD rats were selected and randomized into blank group,model group,celecoxib group,DH-QJ low-dose group,DH-QJ high-dose group,DH-QJ polysaccharide group,and DH-QJ supernatant group,with 8 cases in each group.The rats in blank group were intervened by knee intra-ar-ticular injection of normal saline,while the rest were modeled by knee intra-articular injection of sodium monoiodoacetate to induce KOA.From day 7 after successful modeling,the rats in celecoxib group,DH-QJ low-dose group,DH-QJ high-dose group,DH-QJ polysaccharide group,and DH-QJ supernatant group were intervened by intragastric administration with celecoxib solution(0.018 g/kg),DH-QJ solution(crude drug:2.1 g/kg),DH-QJ solution(crude drug:4.2 g/kg),extracted DH-QJ polysaccharide,and extracted DH-QJ supernatant,re-spectively,while the ones in blank group and model group with the same dose of normal saline,once a day for consecutive 4 weeks.The knee diameter difference(difference between post-modeling diameter and pre-modeling diameter)of the rats in each group were measured before and after the modeling.After the end of the intervention,the rat knee joints and faeces were harvested,and the blood was drawn from the abdominal aorta of rats in each group.The knee joints were stained with HE to observe the histopathological changes,and the degree of cartilage lesions was assessed using the modified Mankin's score.Furthermore,the serum levels of tumor necrosis factor-α(TNF-α),inter-leukin(IL)-6,IL-10,matrix metalloproteinase(MMP)-1,MMP-3 and MMP-13 in the abdominal aortic blood were detected by ELISA,and the microbial communities in the rat fecal samples were identified by 16S ribosomal DNA(rDNA)sequencing.Results:①The knee diameter difference.On day 7 after the modeling,the knee diameter difference was smaller in blank group compared to the other 6 groups(P=0.000,P=0.000,P=0.000,P=0.000,P=0.000,P=0.000).On day 14 after the modeling,the knee diameter difference was smaller in celecoxib group compared to model group(P=0.044).②The histopathological findings in knee joint.The modified Mankin's score was higher in model group compared to blank group,celecoxib group,DH-QJ high-dose group,DH-QJ polysaccharide group,and DH-QJ super-natant group(P=0.000,P=0.003,P=0.003,P=0.000,P=0.000),was higher in celecoxib group compared to DH-QJ high-dose group(P=0.044),and was higher in DH-QJ low-dose group compared to DH-QJ high-dose group,DH-QJ polysaccharide group,and DH-QJ supernatant group(P=0.000,P=0.003,P=0.012).③The serum level of TNF-α.The serum level of TNF-α was lower in blank group,celecoxib group,DH-QJ high-dose group,and DH-QJ supernatant group compared to model group(P=0.038,P=0.016,P=0.016,P=0.016),and was lower in celecoxib group,DH-QJ high-dose group,and DH-QJ supernatant group compared to DH-QJ low-dose group and DH-QJ polysaccharide group(P=0.029,P=0.034;P=0.000,P=0.000;P=0.008,P=0.009).④The serum level of IL-6.The serum level of IL-6 was lower in blank group,celecoxib group,DH-QJ high-dose group,and DH-QJ supernatant group compared to model group(P=0.012,P=0.000,P=0.000,P=0.000),was lower in celecoxib group and DH-QJ supernatant group compared to DH-QJ low-dose group and DH-QJ polysaccharide group(P=0.000,P=0.000;P=0.000,P=0.000),and was lower in DH-QJ high-dose group compared to DH-QJ low-dose group,DH-QJ polysaccharide group,and DH-QJ supernatant group(P=0.000,P=0.000,P=0.028).⑤The serum level of IL-10.The serum level of IL-10 was higher in blank group,celecoxib group,DH-QJ high-dose group,and DH-QJ supernatant group compared to model group(P=0.001,P=0.001,P=0.000,P=0.000),was higher in celecoxib group com-pared to DH-QJ polysaccharide group(P=0.000),was higher in DH-QJ high-dose group and DH-QJ supernatant group compared to DH-QJ low-dose group and DH-QJ polysaccharide group(P=0.010,P=0.000;P=0.000,P=0.000),and was higher in DH-QJ low-dose group compared to DH-QJ polysaccharide group(P=0.004).⑥The serum level of MMP-1.The serum level of MMP-1 was lower in blank group,celecoxib group,DH-QJ high-dose group,and DH-QJ supernatant group compared to model group(P=0.000,P=0.000,P=0.001,P=0.000),was lower in celecoxib group compared to DH-QJ low-dose group and DH-QJ polysaccharide group(P=0.005,P=0.017),and was lower in DH-QJ supernatant group compared to DH-QJ low-dose group,DH-QJ high-dose group,and DH-QJ polysaccha-ride group(P=0.001,P=0.022,P=0.004).⑦The serum level of MMP-3.The serum level of MMP-3 was lower in blank group,celecox-ib group,and DH-QJ supernatant group compared to model group(P=0.041,P=0.003,P=0.000),was lower in celecoxib group com-pared to DH-QJ low-dose group and DH-QJ polysaccharide group(P=0.011,P=0.047),and was lower in DH-QJ supernatant group compared to DH-QJ low-dose group,DH-QJ high-dose group,and DH-QJ polysaccharide group(P=0.000,P=0.005,P=0.000).⑧The serum level of MMP-13.The serum level of MMP-13 was lower in blank group,celecoxib group,DH-QJ high-dose group,and DH-QJ supernatant group compared to model group(P=0.000,P=0.007,P=0.000,P=0.001),and was lower in DH-QJ high-dose group and DH-QJ supernatant group compared to DH-QJ low-dose group and DH-QJ polysaccharide group(P=0.000,P=0.000;P=0.021,P=0.002).⑨The gut microbiota.The Alpha diversity analysis revealed that there was no statistical difference in Chao1 and Shannon indi-ces of gut microbiota among the 7 groups in general.The Beta diversity analysis showed that the inter-group differences in gut microbial composition exceeded the intra-group variations(R2=0.369,P=0.014).The species composition and LEfSe analysis indicated that,at the phylum level,the relative abundance of Firmicutes in the gut microbiota increased,while that of Bacteroidota decreased in DH-QJ high-dose group and DH-QJ supernatant group compared to model group;at the genus level,the relative abundance of norank_f_Muribaculaceae,UCG-005,and Blautia in the gut microbiota increased,while that of norank_o__Clostridia_UCG-014(except for DH-QJ supernatant group)and Romboutsia decreased in DH-QJ low-dose group,DH-QJ high-dose group,DH-QJ polysaccharide group,and DH-QJ supernatant group compared to model group;apart from that,the relative abundance of Lactobacillus in the gut microbiota increased in celecoxib group and DH-QJ high-dose group,and that of Blautia increased in celecoxib group compared to model group.Conclusion:DH-QJ can alleviate the inflammatory response and protect the articular cartilage in KOA model rats,potentially via active components in its supernatant.It may exert the effects by modulating the gut microbiota via enriching the beneficial taxa,e.g.,norank_f_Muribaculaceae,Lactobacillus,UCG-005,and Blautia,and suppressing the pathogenic genera,e.g.,norank_o__Clostridia_UCG-014 and Romboutsia.关键词
骨关节炎,膝/大鼠/独活(中药)/秦艽/胃肠道微生物群/DNA,核糖体Key words
osteoarthritis,knee/rats/angelicae pubescentis radix(TCD)/gentianae macrophyllae radix/gastrointestinal microbiome/DNA,ribosomal引用本文复制引用
赵晨洋,张园媛,周娇,任育鹏,石崇荣,匡海学,王秋红..独活-秦艽对膝骨关节炎模型大鼠的影响及其作用机制研究[J].中医正骨,2025,37(5):30-41,12.基金项目
全国名老中医药专家传承工作室建设项目(国中医药人教函[2022]75号) （国中医药人教函[2022]75号）
第七批全国老中医药专家学术经验继承工作项目(国中医药人教函[2022]76号) （国中医药人教函[2022]76号）
黑龙江省"头雁"团队支持项目(黑龙江省头雁行动领导小组文件[2019]5号) （黑龙江省头雁行动领导小组文件[2019]5号）
中医药传承与创新"百千万"人才工程——岐黄工程首席科学家项目(国中医药人教函[2021]7号) （国中医药人教函[2021]7号）
全国老药工传承工作室建设项目(国中医药人教函[2024]255号) （国中医药人教函[2024]255号）
