中医正骨2024，Vol.36Issue(6) ：10-15,31.

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

Effects of berberine hydrochloride on proliferation and osteogenic differentiation of fibroblast-like synoviocytes derived from patients with ankylosing spondylitis in vitro

刘跃振 张邦能 汪丽娟 孟祥云 张磊 李红专
中医正骨2024，Vol.36Issue(6) ：10-15,31.
✕
  • NETL
  • NSTL
  • 万方数据

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

Effects of berberine hydrochloride on proliferation and osteogenic differentiation of fibroblast-like synoviocytes derived from patients with ankylosing spondylitis in vitro

刘跃振 1张邦能 2汪丽娟 2孟祥云 2张磊 2李红专2
扫码查看

作者信息

  • 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
  • 折叠

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 fi-broblast-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 ho-meostasis regulatory factors were detected by using enzyme-linked immunosorbent assay(ELISA),meanwhile,the cellular osteogenic differ-entiation was observed via alizarin red staining(ARS).Results:① The 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 medi-um-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,com-pared 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 com-pared 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 de-rived 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.

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

Key words

spondylitis,ankylosing/berberine/fibroblast-like synoviocytes/cell proliferation/cell differentiation

引用本文复制引用

基金项目

甘肃省科技重点研发项目(21YF5FA025)

出版年

2024
中医正骨
河南省正骨研究院

中医正骨

CSTPCD
影响因子：1.912
ISSN：1001-6015
参考文献量1
段落导航相关论文