仙龙颗粒联合骨髓间充质干细胞治疗类风湿性关节炎大鼠的疗效及对辅助性T细胞17/调节性T细胞平衡的影响

Efficacy of Xianlong Granule Combined with Bone Mesenchymal Stem Cell in the Treatment of Rheumatoid Arthritis Rats and Its Influence on Th17/Treg Balance

目的探讨仙龙颗粒联合骨髓间充质干细胞(Bone mesenchymal stem cells,BMSC)治疗类风湿性关节炎(Rheumatoid arthritis,RA)的疗效及对辅助性T细胞17(Thelper cell 17,Th17)/调节性T细胞(Tregulatory cells,Treg)平衡的影响。方法将35只8周龄SD大鼠随机分为模型组、泼尼松组、仙龙颗粒组、BMSC组、联合组5组,每组各7只。造模后7 d,模型组、泼尼松组、仙龙颗粒组分别给予生理盐水、泼尼松溶液、仙龙颗粒溶液灌胃;BMSC组、联合组尾静脉注射BMSC后,再分别给予生理盐水、仙龙颗粒灌胃,均连续灌胃4周。治疗后对各组大鼠进行关节炎指数(Arthritis,AI)、足肿胀程度,滑膜组织Th17、Treg细胞,血清肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)、白介素-17(Interleukin-17,IL-17)、白介素-10(Interleukin-10,IL-10)及叉状头/翅膀状螺旋转录因子3(Forkhead/winged helix family transcription factor 3,FoxP3)和维甲酸相关孤儿受体γt(Retinoid-related orphan nuclear receptorγt,RORγt)mRNA水平测量。结果治疗后泼尼松组、仙龙颗粒组、BMSC组和联合组AI评分、足肿胀程度均低于模型组,且联合组均低于泼尼松组、仙龙颗粒组和BMSC组(P<0.05)。治疗后泼尼松组、仙龙颗粒组、BMSC组和联合组滑膜组织Th17、Th17/Treg、RORγt mRNA水平均低于模型组,Treg、FoxP3 mRNA水平均高于模型组,且联合组滑膜组织Th17、Th17/Treg、RORγt mRNA水平均低于泼尼松组、仙龙颗粒组和BMSC组,Treg、FoxP3 mRNA水平均高于泼尼松组、仙龙颗粒组和BMSC组(P<0.05)。治疗后,泼尼松组、仙龙颗粒组、BMSC组和联合组血清TNF-α、IL-17、IL-10水平均低于模型组,且联合组均低于泼尼松组、仙龙颗粒组和BMSC组(P<0.05)。结论仙龙颗粒联合BMSC治疗RA大鼠效果好,能够有效改善其AI评分和足肿胀度,还能够通过调节FoxP3 mRNA、RORγt mRNA水平改善RA大鼠的Th17/Treg细胞平衡状态,进而发挥抗炎作用。

Objective To investigate the efficacy of Xianlong granule combined with bone mesenchymal stem cells(BMSC)in the treatment of rheumatoid arthritis(RA)and its influence on T helper cell 17(Th17)/T regulatory cell(Treg)balance.Methods 35 SD rats aged 8 weeks were randomly divided into 5 groups,with 7 rats in each group.They are model group,prednisone group,Xianlong granule group,BMSC group,and combined group.After 7 days of modeling,the model group,prednisone group and Xianlong granule group were given normal saline,prednisone solution and Xianlong granule solution by gavage respectively.After the BMSC group and the combination group were injected with BMSC via tail vein,they were given normal saline and Xianlong granules by gavage respectively.All patients were given gavage for 4 weeks.After treatment,arthritis index(AI),foot swelling degree,synovial tissue Th17,Treg cells,serum tumor necrosis factor-α(TNF-α),interleukin-17(IL-17),IL-10,synovial tissue forkhead/winged helix family transcription factor 3(Foxp3)and retinoid-related orphan nuclear receptorγt(RORγt)mRNA levels were measured in each group.Results After treatment,AI score and foot swelling degree of prednisone group,Xianlong granule group,BMSC group and combined group were lower than model group,and combined group were lower than prednisone group,Xianlong granule group and BMSC group,and the differences were statistically significant(P<0.05).After treatment,the levels of synovial tissueTh17,Th17/Treg,RORγt mRNA in prednisone group,xianlong granule group,BMSC group and combined group were lower than that in model group,and the levels of Treg and Foxp3 mRNA were higher than that in model group.The levels of synovial tissueTh17,Th17/Treg,RORγt mRNA in combined group were lower than those in prednisone group,xianlong granule group and BMSC group,and the levels of Treg and Foxp3 mRNA in combined group were higher than those in prednisone group,xianlong granule group and BMSC group,and the differences were statistically significant(P<0.05).After treatment,the serum levels of TNF-α,IL-17 and IL-10 in prednisone group,Xianlong granule group,BMSC group and combined group were lower than those in model group,and the combined group were lower than those in prednisone group,Xianlong granule group and BMSC group,and the differences were statistically significant(P<0.05).Conclusions Xianlong granule combined with BMSC has a good effect in the treatment of RA rats,which can effectively improve AI score and foot swelling degree,and improve the balance state of Th17/Treg cells in RA rats by regulating Foxp3 mRNA and RORγt mRNA levels,thus exerting anti-inflammatory effects.