摘要
目的评价山茱萸水提液(CO)、0.05μm无机陶瓷膜微滤液(CO-0.05)及中空10 K纤维膜超滤液(CO-10K)对成纤维样滑膜细胞(HFLS)和佐剂性关节炎(AA)大鼠的抗炎作用,评价膜分离技术富集山茱萸抗炎组分的适用性。方法制备白细胞介素-1β(IL-1β)/肿瘤坏死因子-α(TNF-α)诱导的HFLS炎症模型,CCK-8和ELISA法分别检测CO、CO-0.05及CO-10K对HFLS活力和分泌炎症细胞因子的影响;制备AA大鼠模型,SD大鼠分为对照组、模型组、CO(120 mg/kg)组、CO-0.05(120 mg/kg)组、CO-10K(120 mg/kg)组、阳性对照白芍总苷(TGP,0.125 mg/kg)组,造模同时ig给药,共给药23 d。检测各组大鼠足趾肿胀、体质量,ELISA法测定大鼠血清中炎症因子IL-1、IL-6、前列腺素E_2(PGE_2)、TNF-α水平。结果 CO、CO-0.05及CO-10K均可以显著抑制IL-1β/TNF-α诱导的滑膜细胞分泌细胞因子(P<0.05、0.01、0.001),并且能够显著缓解AA大鼠足趾肿胀(P<0.05、0.01、0.001),改善大鼠体质量增长缓慢的情况(P<0.05、0.01、0.001),减少AA大鼠血清中炎性细胞因子的产生(P<0.001)。通过山茱萸膜微滤液和膜超滤液对炎症细胞因子产生的抑制率比较,发现膜超滤液具有更显著的抑制炎症因子产生的作用。结论 CO-0.05和CO-10K均具有显著的抗炎作用,而中空纤维膜超滤分离技术更适用于山茱萸抗炎组分的富集。
Objective To investigate the anti-inflammatory effect of Cornus officinalis(CO) Decotion and its refined solutions from membrane separation by using 0.05 μm inorganic ceramic membrane(CO-0.05) and 10 K polysulfone hollow fiber membrane(CO-10 K), and evaluate the applicability of the membrane separation technique for concentrating the anti-inflammatory compounds of C. officinalis Decotion. Methods Inflammatory model of interleukin(IL)-1β and tumor necrosis factor(TNF)-α stimulated human fibroblast-like synoviocytes(HFLS) was prepared. The CCK-8 assay and ELISA were applied to detect the effects of C. officinalis Decotion and its refined solutions on the viability of HFLS and the secretion of inflammatory cytokines, respectively. Moreover, the animal model of adjuvant arthritis(AA) was used. The SD rats were divided into six groups: control group, model(AA) group and AA groups intragastrically receiving CO(120 mg/kg), CO-0.05(120 mg/kg), CO-10 K(120 mg/kg) and TGP(0.125 mg/kg) with daily treatments for 23 days. The weight and paw swelling of rats in different groups were detected. The ELISA was used to detect secretion levels of prostaglandin E_2(PGE_2), IL-1, IL-6, and TNF-α in serum. Results The production of vascular endothelial growth factor(VEGF) induced by IL-1β/TNF-α were significantly inhibited with C. officinalis Decotion and its refined solutions by membrane separation treatment(P < 0.05, 0.01, 0.001). C. officinalis Decoction and the refined solutions significantly ameliorated paw swelling and increased weight gain of AA rats(P < 0.05, 0.01, 0.001), and reduced the secretion of TNF-α, PGE_2, IL-1, IL-6 in serum(P < 0.001). By comparing the inhibition efficiency of inflammatory cytokines by inorganic ceramic membrane refined solution and polysulfone hollow fiber membrane refined solution, the polysulfone hollow fiber membrane refined solution exhibited better anti-inflammatory activity. Conclusion Both of refined solutions of C. officinalis Decotion from inorganic ceramic membrane and polysulfone hollow fiber membrane separation exhibited dramatically anti-inflammtory activity. Moreover, the polysulfone hollow fiber membrane was more applicable for concentrating the anti-inflammatory compounds of C. officinalis Decotion.
引文
[1]Calabresi E,Petrelli F F,Bonifacio A,et al.One year in review 2018:Pathogenesis of rheumatoid arthritis[J].Clin Exp Rheumatol,2018,36(1):175-184.
[2]魏志萍,洪芬芳,杨树龙.中药治疗类风湿关节炎的研究进展[J].中华中医药杂志,2017,32(12):5477-5481.
[3]池里群,周彬,高文远,等.治疗类风湿性关节炎常用药物的研究进展[J].中国中药杂志,2014,39(15):2851-2858.
[4]Jagpal A,Curtis J R.Gastrointestinal perforations with piologics in patients with rheumatoid arthritis:Implications for clinicians[J].Drug Safety,2018:doi:10.1007/s40264-018-0639-1.
[5]Bartok B,Firestein G S.Fibroblast-like synoviocytes:Key effector cells in rheumatoid arthritis[J].Immunol Rev,2010,233(1):233-255.
[6]Ganesan R,Rasool M.Fibroblast-like synoviocytesdependent effector molecules as a critical mediator for rheumatoid arthritis:Current status and future directions[J].Inter Rev Immunol,2017,36(1):20-30.
[7]Kaneko S,Kondo Y,Yokosawa M,et al.Rheumatoid arthritis and cytokines[J].J Clin Med,2016,74(6):913-918.
[8]Meng Q L,Du X Z,Wang H L,et al.Astragalus polysaccharides inhibits cell growth and proinflammatory response in IL-1β-stimulated fibroblast-like synoviocytes by enhancement of autophagy via PI3K/AKT/m TOR inhibition[J].Apoptosis,2017,22(9):1138-1146.
[9]Rebrov B,Komarova E,Blagodarenko A,et al.AB0196Vascular endothelial growth factor in patients with rheumatoid arthritis[J].Ann Rheum Dis,2017,76(2):1115-1116.
[10]岑琴,周丽莉,礼彤.膜分离技术及其在中药领域中的应用[J].沈阳药科大学学报,2008,25(1):77-80.
[11]陈伟平.膜分离技术分离昆明山海棠根提取液总生物碱[J].中药材,2014,37(7):1287-1289.
[12]刘红波,唐志书,宋忠兴,等.膜分离技术对沙棘籽油精制的初步研究[J].膜科学与技术,2017,37(2):124-130.
[13]欧阳丽,何鼎胜,苏孝礼,等.反萃分散液膜分离提取黄连中生物碱研究[J].分析化学,2009,37(3):346-346.
[14]叶贤胜,赫军,张佳琳,等.山茱萸的化学成分研究[J].中国中药杂志,2016,41(24):4605-4609.
[15]杨明明,袁晓旭,赵桂琴,等.山茱萸化学成分和药理作用的研究进展[J].承德医学院学报,2016,33(5):398-400.
[16]武婧,唐志书,古川,等.无机陶瓷膜微滤精制山茱萸水提液的工艺研究[J].中南药学,2015,13(9):943-946.
[17]武婧,唐志书,宋忠兴,等.山茱萸水提液超滤工艺优化研究[J].中草药,2015,46(22):3338-3343.
[18]武婧.膜分离技术富集山茱萸抗风湿组分的适用性研究与评价[D].西安:陕西中医药大学,2016.
[19]李建民.山茱萸总甙抗类风湿性关节炎分子免疫机理研究[D].北京:北京中医药大学,2001.
[20]宋逍,段玺,赵鹏,等.膜分离技术应用于柴黄口服液的纯化分离工艺研究[J].吉林中医药,2017,37(2):183-187.
[21]张可人,张依人,彭买姣,等.陶瓷膜分离技术精制古汉养生精提取液的工艺研究[J].湖南中医药大学学报,2018,38(8):875-878.
[22]朱华旭,唐志书,段金廒,等.基于资源循环经济的中药脉络宁注射液废弃物资源化循环利用中的共性关键问题[J].中国现代中药,2017,19(12):1672-1687.
