[1]刘跃振,张邦能,汪丽娟,等.盐酸小檗碱对强直性脊柱炎患者成纤维样滑膜细胞体外增殖和成骨分化的影响[J].中医正骨,2024,36(06):10-15，31.
　LIU Yuezhen,ZHANG Bangneng,WANG Lijuan,et al.Effects of berberine hydrochloride on proliferation and osteogenic differentiation of fibroblast-like synoviocytes derived from patients with ankylosing spondylitis in vitro[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2024,36(06):10-15，31.
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盐酸小檗碱对强直性脊柱炎患者成纤维样滑膜细胞体外增殖和成骨分化的影响()
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《中医正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第36卷
期数:
2024年06期
页码:
10-15，31
栏目:
基础研究
出版日期:
2024-06-20

文章信息/Info

Title:
Effects of berberine hydrochloride on proliferation and osteogenic differentiation of fibroblast-like synoviocytes derived from patients with ankylosing spondylitis in vitro
作者:
刘跃振1; 张邦能2; 汪丽娟2; 孟祥云2; 张磊2; 李红专2
1.甘肃中医药大学,甘肃 兰州 730000; 2.甘肃省中医院,甘肃 兰州 730050
Author(s):
LIU Yuezhen1; ZHANG Bangneng2; WANG Lijuan2; MENG Xiangyun2; ZHANG Lei2;  LI Hongzhuan2
1.Gansu University of Chinese Medicine,Lanzhou 730000,Gansu,China 2.Gansu Provincial Hospital of TCM,Lanzhou 730050,Gansu,China
关键词:
脊柱炎; 强直性;  小檗碱;  成纤维样滑膜细胞;  细胞增殖;  细胞分化
Keywords:
spondylitis; ankylosing;  berberine;  fibroblast-like synoviocytes;  cell proliferation;  cell differentiation
摘要:
目的:探讨盐酸小檗碱对强直性脊柱炎(ankylosing spondylitis,AS)患者成纤维样滑膜细胞(fibroblast-like synoviocytes,FLS)体外增殖和成骨分化的影响。方法:从AS患者的髋关节滑膜组织中分离培养FLS,经免疫荧光染色鉴定后进行后续实验。取第3代FLS,以白细胞介素(interleukin,IL)-17、IL-23预处理,再以0 μmol·L-1(空白组)、80 μmol·L-1(盐酸小檗碱低剂量组)、120 μmol·L-1(盐酸小檗碱中剂量组)、200 μmol·L-1(盐酸小檗碱高剂量组)的盐酸小檗碱干预后,采用光学显微镜观察细胞增殖情况,以酶联免疫吸附法检测炎症因子水平和骨稳态调节因子水平,通过茜素红染色观察细胞成骨分化情况。结果:①炎症因子水平测定结果。盐酸小檗碱低、中、高剂量组IL-17、IL-23、IL-6、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)水平均低于空白组(IL-17:P=0.046,P=0.005,P=0.004; IL-23:P=0.024,P=0.002,P=0.000; IL-6:P=0.012,P=0.000,P=0.000; TNF-α:P=0.012,P=0.004,P=0.001),盐酸小檗碱中、高剂量组IL-17、IL-23、IL-6、TNF-α水平均低于盐酸小檗碱低剂量组(IL-17:P=0.034,P=0.024; IL-23:P=0.020,P=0.000; IL-6:P=0.000,P=0.000; TNF-α:P=0.012,P=0.002),盐酸小檗碱高剂量组IL-17、IL-23、IL-6、TNF-α水平均低于盐酸小檗碱中剂量组(IL-17:P=0.029; IL-23:P=0.014; IL-6:P=0.005; TNF-α:P=0.026)。②FLS增殖情况观察结果。光学显微镜观察结果显示,与空白组相比,盐酸小檗碱低、中、高剂量组细胞间隙增大、凋亡细胞数量增加,变化程度呈现剂量依赖性。③FLS成骨分化情况观察结果。与空白组相比,盐酸小檗碱低、中、高剂量组茜素红染色程度均降低,而且降低程度呈现剂量依赖性。④骨稳态调节因子水平测定结果。盐酸小檗碱低、中、高剂量组Dickkopf相关蛋白1(dickkopf-related protein 1,DKK-1)和核因子κB受体激活蛋白配体(receptor activator of NF-κB ligand,RANKL)水平均低于空白组(DKK-1:P=0.048,P=0.032,P=0.001; RANKL:P=0.046,P=0.021,P=0.001),盐酸小檗碱中、高剂量组DKK-1和RANKL水平均低于盐酸小檗碱低剂量组(DKK-1:P=0.028,P=0.002; RANKL:P=0.047,P=0.002),盐酸小檗碱高剂量组DKK-1和RANKL水平均低于盐酸小檗碱中剂量组(DKK-1:P=0.018; RANKL:P=0.011)。结论:盐酸小檗碱能够抑制AS患者FLS体外增殖,并通过减少FLS分泌骨稳态调节因子RANKL和DKK-1抑制FLS成骨分化,而且其作用具有剂量依赖性。
Abstract:
Objective:To investigate the effects of berberine hydrochloride(BBH)on proliferation and osteogenic differentiation of fibroblast-like synoviocytes(FLSs)derived from patients with ankylosing spondylitis(AS)in vitro.Methods:The FLSs were isolated from the hip synovial tissues of AS patients and cultured,and then identified by immunofluorescence staining for subsequent experiments.The third-generation FLSs were collected and pretreated with interleukin(IL)-17 and IL-23,followed by intervention with BBH at concentrations of 0(blank group),80(low-dose BBH group),120(medium-dose BBH group)and 200 μmol/L(high-dose BBH group),respectively.After the end of intervention,the cellular proliferation was observed by using optical microscopy,and the levels of inflammatory factors and bone homeostasis regulatory factors were detected by using enzyme-linked immunosorbent assay(ELISA),meanwhile,the cellular osteogenic differentiation was observed via alizarin red staining(ARS).Results:①The levels of IL-17,IL-23,IL-6 and tumor necrosis factor-α(TNF-α)were lower in low-,medium-,and high-dose BBH group compared to blank group(IL-17:P=0.046,P=0.005,P=0.004; IL-23:P=0.024,P=0.002,P=0.000; IL-6:P=0.012,P=0.000,P=0.000; TNF-α:P=0.012,P=0.004,P=0.001),and were lower in medium- and high-dose BBH group compared to low-dose BBH group(IL-17:P=0.034,P=0.024; IL-23:P=0.020,P=0.000; IL-6:P=0.000,P=0.000; TNF-α:P=0.012,P=0.002),and were lower in high-dose BBH group compared to medium-dose BBH group(IL-17:P=0.029; IL-23:P=0.014; IL-6:P=0.005; TNF-α:P=0.026).②The results of observation by optical microscopy showed that,compared to blank group,the intercellular space and the number of apoptotic cells increased in low-,medium-,and high-dose BBH group,with a dose-dependence exhibited in the degree of change.③Compared with that of blank group,the ARS intensity was reduced in low-,medium-,and high-dose BBH group,with a dose-dependence exhibited in the degree of reduction.④The levels of Dickkopf-related protein 1(DKK1)and receptor activator of NF-κB ligand(RANKL)were lower in low-,medium-,and high-dose BBH group compared to blank group(DKK1:P=0.048,P=0.032,P=0.001; RANKL:P=0.046,P=0.021,P=0.001),and were lower in medium- and high-dose BBH group compared to low-dose BBH group(DKK1:P=0.028,P=0.002; RANKL:P=0.047,P=0.002),and were lower in high-dose BBH group compared to medium-dose BBH group(DKK1:P=0.018; RANKL:P=0.011).Conclusion:BBH can inhibit the proliferation of FLSs derived from AS patients in vitro,and repress the osteogenic differentiation of FLSs via reducing the bone homeostasis regulatory factors RANKL and DKK1 secreted by FLSs,with a dose-dependence exhibited in the effects.

参考文献/References:

[1] MCGONAGLE D,DAVID P,MACLEOD T,et al.Predominant ligament-centric soft-tissue involvement differentiates axial psoriatic arthritis from ankylosing spondylitis[J].Nat Rev Rheumatol,2023,19(12):818-827.
[2] ZHANG S,XU P,ZHU Z,et al.Acetylation of p65(Lys310)by p300 in macrophages mediates anti-inflammatory property of berberine[J].Redox Biol,2023,62:102704.
[3] SAMADI P,SARVARIAN P,GHOLIPOUR E,et al.Berbe-rine:a novel therapeutic strategy for cancer[J].IUBMB Life,2020,72(10):2065-2079.
[4] BLAIS J E,HUANG X,ZHAO J V.Overall and sex-specific effect of berberine for the treatment of dyslipidemia in adults:a systematic review and meta-analysis of randomized placebo-controlled trials[J].Drugs,2023,83(5):403-427.
[5] CORTEZ-NAVARRETE M,PÉREZ-RUBIO K G,ESCOBEDO-GUTIÉRREZ M J.Role of fenugreek,cinnamon,curcuma longa,berberine and momordica charantia in type 2 diabetes mellitus treatment:a review[J].Pharmaceuticals(Basel),2023,16(4):515.
[6] YANG M,YANG T,MEI L,et al.The potential of berberine to target telocytes in rabbit heart[J].Planta Med,2024,90(2):84-95.
[7] SUADONI M T,ATHERTON I.Berberine for the treatment of hypertension:a systematic review[J].Complement Ther Clin Pract,2021,42:101287.
[8] MA W,ZHANG Y,YU M,et al.In-vitro and in-vivo anti-breast cancer activity of synergistic effect of berberine and exercise through promoting the apoptosis and immunomodulatory effects[J].Int Immunopharmacol,2020,87:106787.
[9] SHEN P,JIAO Y,MIAO L,et al.Immunomodulatory effects of berberine on the inflamed joint reveal new therapeutic targets for rheumatoid arthritis management[J].J Cell Mol Med,2020,24(21):12234-12245.
[10] EHTESHAMFAR S M,AKHBARI M,AFSHARI J T,et al.Anti-inflammatory and immune-modulatory impacts of berberine on activation of autoreactive T cells in autoimmune inflammation[J].J Cell Mol Med,2020,24(23):13573-13588.
[11] SINGH S,PATHAK N,FATIMA E,et al.Plant isoquinoline alkaloids:advances in the chemistry and biology of berbe-rine[J].Eur J Med Chem,2021,226:113839.
[12] OCH A,PODGÓRSKI R,NOWAK R.Biological activity of berberine—a summary update[J].Toxins(Basel),2020,12(11):713.
[13] VAN DER LINDEN S,VALKENBURG H A,CATS A.Evaluation of diagnostic criteria for ankylosing spondylitis.A proposal for modification of the New York criteria[J].Arthritis Rheum,1984,27(4):361-368.
[14] TSUKAZAKI H,KAITO T.The role of the IL-23/IL-17 pathway in the pathogenesis of spondyloarthritis[J].Int J Mol Sci,2020,21(17):6401.
[15] HAONAN L,ZEHANG S,JIACONG H,et al.Interleukin-23 mediates the reduction of GADD45a expression to attenuate oxidative stress-induced cellular senescence in human fibroblasts[J].Mech Ageing Dev,2023,212:111808.
[16] DING M,CHENG Y,XU Z,et al.Hypoxia inhibits osteogenesis and promotes adipogenesis of fibroblast-like synoviocytes via upregulation of leptin in patients with rheumatoid arthritis[J].J Immunol Res,2022(1):1431399.
[17] WEI L,ZHANG X,YAO Y,et al.LncRNA HOTTIP impacts the proliferation and differentiation of fibroblast-like synoviocytes in ankylosing spondylitis through the microRNA-30b-3p/PGK1 axis[J].J Orthop Surg Res,2023,18(1):237.
[18] NYGAARD G,FIRESTEIN G S.Restoring synovial homeostasis in rheumatoid arthritis by targeting fibroblast-like synoviocytes[J].Nat Rev Rheumatol,2020,16(6):316-333.
[19] 叶倩仪,牟兴,吴歆,等.成纤维细胞在自身免疫性疾病中作用的研究进展[J].现代免疫学,2022,42(5):421-426.
[20] YU Y,CAI W,ZHOU J,et al.Anti-arthritis effect of berbe-rine associated with regulating energy metabolism of macrophages through AMPK/HIF-1α pathway[J].Int Immunopharmacol,2020,87:106830.
[21] DINESH P,RASOOL M.Berberine mitigates IL-21/IL-21R mediated autophagic influx in fibroblast-like synoviocytes and regulates Th17/Treg imbalance in rheumatoid arthritis[J].Apoptosis,2019,24(7/8):644-661.
[22] BIEDRON G,CZEPIEL M,SIEDLAR M,et al.Serum concentration of dickkopf-related protein 1(DKK1)in psoriatic arthritis in the context of bone remodelling[J].Rheumatol Int,2023,43(12):2175-2183.
[23] CHEN S,LI Z,CHEN D,et al.Piezo1-mediated mechanotransduction promotes entheseal pathological new bone formation in ankylosing spondylitis[J].Ann Rheum Dis,2023,82(4):533-545.
[24] UCIECHOWSKI P,DEMPKE W.Interleukin-6:a masterplayer in the cytokine network[J].Oncology,2020,98(3):131-137.
[25] WANG Q,LIU Y,WU J,et al.Potential significance of changes in serum levels of IL-17,TNF-α and DKK-1 in the progression of the rheumatoid arthritis[J].Autoimmunity,2023,56(1):2276068.
[26] ZHANG L,OUYANG H,XIE Z,et al.Serum DKK-1 level in the development of ankylosing spondylitis and rheumatic arthritis:a meta-analysis[J].Exp Mol Med,2016,48(4):e228.

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备注/Memo

备注/Memo:
基金项目:甘肃省科技重点研发项目(21YF5FA025)
通讯作者:张邦能 E-mail:zbn81@163.com
更新日期/Last Update: 1900-01-01