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[1]劉躍振,張邦能,汪麗娟,等.鹽酸小檗堿對強直性脊柱炎患者成纖維樣滑膜細胞體外增殖和成骨分化的影響[J].中醫(yī)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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鹽酸小檗堿對強直性脊柱炎患者成纖維樣滑膜細胞體外增殖和成骨分化的影響()
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《中醫(yī)正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第36卷
期數(shù):
2024年06期
頁碼:
10-15，31
欄目:
基礎研究
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:
劉躍振1; 張邦能2; 汪麗娟2; 孟祥云2; 張磊2; 李紅專2
1.甘肅中醫(yī)藥大學,甘肅 蘭州 730000; 2.甘肅省中醫(yī)院,甘肅 蘭州 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
關鍵詞:
脊柱炎; 強直性;  小檗堿;  成纖維樣滑膜細胞;  細胞增殖;  細胞分化
Keywords:
spondylitis; ankylosing;  berberine;  fibroblast-like synoviocytes;  cell proliferation;  cell differentiation
摘要:
目的:探討鹽酸小檗堿對強直性脊柱炎(ankylosing spondylitis,AS)患者成纖維樣滑膜細胞(fibroblast-like synoviocytes,FLS)體外增殖和成骨分化的影響。方法:從AS患者的髖關節(jié)滑膜組織中分離培養(yǎng)FLS,經(jīng)免疫熒光染色鑒定后進行后續(xù)實驗。取第3代FLS,以白細胞介素(interleukin,IL)-17、IL-23預處理,再以0 μmol·L-1(空白組)、80 μmol·L-1(鹽酸小檗堿低劑量組)、120 μmol·L-1(鹽酸小檗堿中劑量組)、200 μmol·L-1(鹽酸小檗堿高劑量組)的鹽酸小檗堿干預后,采用光學顯微鏡觀察細胞增殖情況,以酶聯(lián)免疫吸附法檢測炎癥因子水平和骨穩(wěn)態(tài)調(diào)節(jié)因子水平,通過茜素紅染色觀察細胞成骨分化情況。結果:①炎癥因子水平測定結果。鹽酸小檗堿低、中、高劑量組IL-17、IL-23、IL-6、腫瘤壞死因子α(tumor necrosis factor-α,TNF-α)水平均低于空白組(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),鹽酸小檗堿中、高劑量組IL-17、IL-23、IL-6、TNF-α水平均低于鹽酸小檗堿低劑量組(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),鹽酸小檗堿高劑量組IL-17、IL-23、IL-6、TNF-α水平均低于鹽酸小檗堿中劑量組(IL-17:P=0.029; IL-23:P=0.014; IL-6:P=0.005; TNF-α:P=0.026)。②FLS增殖情況觀察結果。光學顯微鏡觀察結果顯示,與空白組相比,鹽酸小檗堿低、中、高劑量組細胞間隙增大、凋亡細胞數(shù)量增加,變化程度呈現(xiàn)劑量依賴性。③FLS成骨分化情況觀察結果。與空白組相比,鹽酸小檗堿低、中、高劑量組茜素紅染色程度均降低,而且降低程度呈現(xiàn)劑量依賴性。④骨穩(wěn)態(tài)調(diào)節(jié)因子水平測定結果。鹽酸小檗堿低、中、高劑量組Dickkopf相關蛋白1(dickkopf-related protein 1,DKK-1)和核因子κB受體激活蛋白配體(receptor activator of NF-κB ligand,RANKL)水平均低于空白組(DKK-1:P=0.048,P=0.032,P=0.001; RANKL:P=0.046,P=0.021,P=0.001),鹽酸小檗堿中、高劑量組DKK-1和RANKL水平均低于鹽酸小檗堿低劑量組(DKK-1:P=0.028,P=0.002; RANKL:P=0.047,P=0.002),鹽酸小檗堿高劑量組DKK-1和RANKL水平均低于鹽酸小檗堿中劑量組(DKK-1:P=0.018; RANKL:P=0.011)。結論:鹽酸小檗堿能夠抑制AS患者FLS體外增殖,并通過減少FLS分泌骨穩(wěn)態(tài)調(diào)節(jié)因子RANKL和DKK-1抑制FLS成骨分化,而且其作用具有劑量依賴性。
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.

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備注/Memo

備注/Memo:
基金項目:甘肅省科技重點研發(fā)項目(21YF5FA025)
通訊作者:張邦能 E-mail:[email protected]
更新日期/Last Update: 1900-01-01