[1]于兰,解冬千,王霞,等.蒺藜皂苷对白细胞介素-1β诱导的软骨细胞凋亡和炎症因子分泌的影响[J].中医正骨,2024,36(1):23-32.
　YU Lan,XIE Dongqian,WANG Xia,et al.Effects of tribulus terrestris saponins on interleukin-1β-induced chondrocyte apoptosis and inflammatory factor secretion[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2024,36(1):23-32.
点击复制

蒺藜皂苷对白细胞介素-1β诱导的软骨细胞凋亡和炎症因子分泌的影响()
分享到：

《中医正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第36卷
期数:
2024年1期
页码:
23-32
栏目:
基础研究
出版日期:
2024-01-20

文章信息/Info

Title:
Effects of tribulus terrestris saponins on interleukin-1β-induced chondrocyte apoptosis and inflammatory factor secretion
作者:
于兰; 解冬千; 王霞; 及超
保定市第二医院,河北 保定 071001
Author(s):
YU Lan; XIE Dongqian; WANG Xia; JI Chao
The No.2 Hospital of Baoding,Baoding 071001,Hebei,China
关键词:
骨关节炎;  软骨细胞;  细胞凋亡;  蒺藜;  皂苷类;  白细胞介素-1β;  RNA; 环状;  炎症介导素类
Keywords:
osteoarthritis;  chondrocytes;  apoptosis;  fructus tribuli;  saponins;  interleukin-1 beta;  RNA; circular;  inflammation mediators
摘要:
目的:观察蒺藜皂苷对白细胞介素-1β(interleukin-1β,IL-1β)诱导的软骨细胞凋亡和炎症因子分泌的影响,并探讨其作用机制。方法:①软骨细胞培养和转染。采用DMEM培养液培养小鼠软骨细胞(ATDC5细胞),培养后采用转染试剂分别转染pcDNA-circ_0045714、pcDNA、si-circ_0045714、si-NC。②蒺藜皂苷浓度筛选。将ATDC5细胞接种于96孔板中,一组正常培养(正常组),一组加入10 ng·mL-1的IL-1β(IL-1β组),其他组在加入10 ng·mL-1 IL-1β的基础上分别加入浓度为25 μg·mL-1、50 μg·mL-1、100 μg·mL-1、200 μg·mL-1、400 μg·mL-1的蒺藜皂苷。培养后计算软骨细胞存活率,并确定后续实验蒺藜皂苷的浓度。③软骨细胞增殖抑制率检测。将ATDC5细胞分为对照组、IL-1β组、IL-1β+蒺藜皂苷低剂量组、IL-1β+蒺藜皂苷中剂量组、IL-1β+蒺藜皂苷高剂量组,其中对照组采用常规培养液培养,IL-1β组采用含10 ng·mL-1的IL-1β培养液培养,IL-1β+蒺藜皂苷低、中、高剂量组分别采用含50 μg·mL-1、100 μg·mL-1、200 μg·mL-1的蒺藜皂苷和10 ng·mL-1的IL-1β培养液培养。转染pcDNA-circ_0045714、pcDNA的ATDC5细胞,培养方法同IL-1β组,并分别标记为IL-1β+pcDNA-circ_0045714组、IL-1β+pcDNA组。转染si-circ_0045714、si-NC的ATDC5细胞,培养方法同IL-1β+蒺藜皂苷高剂量组,并分别标记为IL-1β+蒺藜皂苷高剂量+si-circ_0045714组、IL-1β+蒺藜皂苷高剂量+si-NC组。培养后计算软骨细胞增殖抑制率。④软骨细胞凋亡率检测。于6孔板中接种ATDC5细胞及转染pcDNA-circ_0045714、pcDNA、si-circ_0045714、si-NC的ATDC5细胞,培养后采用流式细胞仪检测各组软骨细胞凋亡率。⑤软骨细胞中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和白细胞介素6(interleukin 6,IL-6)含量检测。收集各组软骨细胞的培养液,检测上清液中TNF-α和IL-6含量。⑥软骨细胞中circ_0045714表达量检测。提取各组软骨细胞中总RNA,逆转录合成cDNA,然后扩增,最后计算circ_0045714的表达量。结果:①蒺藜皂苷浓度筛选结果。IL-1β组的软骨细胞存活率低于正常组(LSD-t=15.396,P=0.000)。IL-1β组与IL-1β+25 μg·mL-1蒺藜皂苷组的软骨细胞存活率比较,差异无统计学意义(LSD-t=1.555,P=0.918)。IL-1β+50 μg·mL-1蒺藜皂苷组、IL-1β+100 μg·mL-1蒺藜皂苷组、IL-1β+200 μg·mL-1蒺藜皂苷组、IL-1β+400 μg·mL-1蒺藜皂苷组的软骨细胞存活率均高于IL-1β组(LSD-t=4.879,P=0.047; LSD-t=7.686,P=0.001; LSD-t=10.657,P=0.000; LSD-t=10.073,P=0.000)。IL-1β+400 μg·mL-1蒺藜皂苷组与IL-1β+200 μg·mL-1蒺藜皂苷组的软骨细胞存活率比较,差异无统计学意义(LSD-t=0.584,P=0.999)。因此,选择浓度为50 μg·mL-1、100 μg·mL-1、200 μg·mL-1的蒺藜皂苷进行实验,并分别标记为蒺藜皂苷低剂量组、中剂量组、高剂量组。②软骨细胞增殖抑制率、软骨细胞凋亡率、软骨细胞中TNF-α和IL-6含量检测结果。IL-1β组的软骨细胞增殖抑制率、软骨细胞凋亡率、软骨细胞中TNF-α和IL-6含量均高于对照组、IL-1β+蒺藜皂苷低剂量组、IL-1β+蒺藜皂苷中剂量组、IL-1β+蒺藜皂苷高剂量组(软骨细胞增殖抑制率:LSD-t=55.829,P=0.000; LSD-t=8.879,P=0.001; LSD-t=20.507,P=0.000; LSD-t=30.315,P=0.000; 软骨细胞凋亡率:LSD-t=27.508,P=0.000; LSD-t=5.076,P=0.032; LSD-t=11.689,P=0.000; LSD-t=21.284,P=0.000; 软骨细胞中TNF-α含量:LSD-t=29.990,P=0.000; LSD-t=7.720,P=0.002; LSD-t=17.182,P=0.000; LSD-t=24.615,P=0.000; 软骨细胞中IL-6含量:LSD-t=33.441,P=0.000; LSD-t=6.324,P=0.008; LSD-t=15.440,P=0.000; LSD-t=25.096,P=0.000)。IL-1β+蒺藜皂苷低剂量组的软骨细胞增殖抑制率、软骨细胞凋亡率、软骨细胞中TNF-α和IL-6含量均高于IL-1β+蒺藜皂苷中剂量组、IL-1β+蒺藜皂苷高剂量组(软骨细胞增殖抑制率:(LSD-t=11.627,P=0.000; LSD-t=21.436,P=0.000; 软骨细胞凋亡率:LSD-t=6.613,P=0.006; LSD-t=16.209,P=0.000; 软骨细胞中TNF-α含量:LSD-t=9.463,P=0.000; LSD-t=16.895,P=0.000; 软骨细胞中IL-6含量:LSD-t=9.117,P=0.001; LSD-t=18.773,P=0.000)。IL-1β+蒺藜皂苷中剂量组的软骨细胞增殖抑制率、软骨细胞凋亡率、软骨细胞中TNF-α和IL-6含量均高于IL-1β+蒺藜皂苷高剂量组(LSD-t=9.808,P=0.000; LSD-t=9.595,P=0.000; LSD-t=7.432,P=0.003; LSD-t=9.656,P=0.000)。③软骨细胞中circ_0045714表达量检测结果。IL-1β组软骨细胞中circ_0045714表达量低于对照组、IL-1β+蒺藜皂苷低剂量组、IL-1β+蒺藜皂苷中剂量组、IL-1β+蒺藜皂苷高剂量组(LSD-t=43.218,P=0.000; LSD-t=9.487,P=0.000; LSD-t=22.136,P=0.000; LSD-t=34.785,P=0.000)。IL-1β+蒺藜皂苷低剂量组软骨细胞中circ_0045714表达量低于IL-1β+蒺藜皂苷中剂量组、IL-1β+蒺藜皂苷高剂量组(LSD-t=12.649,P=0.000; LSD-t=25.298,P=0.000)。IL-1β+蒺藜皂苷中剂量组软骨细胞中circ_0045714表达量低于IL-1β+蒺藜皂苷高剂量组(LSD-t=12.649,P=0.000)。④过表达和干扰circ_0045714的软骨细胞构建结果。转染pcDNA、pcDNA-circ_0045714的软骨细胞中circ_0045714表达量比较,差异有统计学意义(1.00±0.00,4.18±0.14,t=39.342,P=0.000),说明过表达circ_0045714的软骨细胞构建成功。转染si-NC、si-circ_0045714的软骨细胞中circ_0045714表达量比较,差异有统计学意义(1.00±0.00,0.28±0.04,t=31.177,P=0.000),说明干扰circ_0045714的软骨细胞构建成功。⑤过表达circ_0045714对IL-1β诱导的软骨细胞影响结果。IL-1β+pcDNA-circ_0045714组的软骨细胞增殖抑制率、软骨细胞凋亡率、软骨细胞中TNF-α和IL-6含量均低于IL-1β+pcDNA组(t=16.290,P=0.000; t=11.359,P=0.000; t=11.988,P=0.000; t=12.266,P=0.000)。⑥干扰circ_0045714对IL-1β诱导的软骨细胞影响结果。IL-1β+蒺藜皂苷高剂量+si-circ_0045714组软骨细胞增殖抑制率、软骨细胞凋亡率、软骨细胞中TNF-α和IL-6含量均高于IL-1β+蒺藜皂苷高剂量+si-NC组(t=9.586,P=0.001; t=9.120,P=0.001; t=7.069,P=0.002; t=10.548,P=0.001)。结论:蒺藜皂苷能抑制IL-1β诱导的软骨细胞凋亡及炎症因子表达,具有治疗骨关节炎的潜在价值,其作用机制可能与上调软骨细胞中circ_0045714表达有关,且200 μg·mL-1蒺藜皂苷的效果更佳。
Abstract:
Objective:To observe the effects of tribulus terrestris saponins(TTS)on interleukin-1β(IL-1β)-induced chondrocyte apoptosis and inflammatory cytokine secretion,and to explore its underlying mechanism.Methods:①The mouse chondrocytes(ATDC5 cells)were harvested and cultured in the Dulbecco's Modified Eagle's Medium(DMEM).After 24-hour culture,the ATDC5 cells were transfected with pcDNA-circ_0045714,pcDNA,si-circ_0045714,and si-NC,respectively,by using transfection reagents.②The ATDC5 cells were inoculated onto the 96-well plates,with one group cultured in normal DMEM(normal group),one group in DMEM containing 10 ng/mL IL-1β(IL-1β group),and the rest in DMEM adding with 10 ng/mL IL-1β and TTS with concentration of 25,50,100,200,and 400 μg/mL,respectively.After 24-hour culture,the survival rate of chondrocytes in each group was calculated,based on which the concentration of TTS used in subsequent experiments was determined.③The ATDC5 cells were divided into control group,IL-1β group,IL-1β+low-dose TTS(L-TTS)group,IL-1β+medium-dose TTS(M-TTS)group,and IL-1β+high-dose TTS(H-TTS)group.The ATDC5 cells in control group were cultured in conventional DMEM,the ones in IL-1β group in DMEM with 10 ng/mL IL-1β,and the ones in the other 3 groups in DMEM adding with 10 ng/mL IL-1β and TTS with concentration of 50,100,and 200 μg/mL,respectively.The pcDNA-circ_0045714- and pcDNA-transfected ATDC5 cells were cultured with the same method as IL-1β group,and were labeled as IL-1β+pcDNA-circ_0045714 group and IL-1β+pcDNA group,respectively.The si-circ_0045714- and si-NC-transfected ATDC5 cells were cultured with the same method as IL-1β+H-TTS group,and were labeled as IL-1β+H-TTS+si-circ_0045714 group and IL-1β+H-TTS+si-NC group,respectively.After 24-hour culture,the proliferation inhibition rate of the chondrocytes in each group was calculated.④The ATDC5 cells and the pcDNA-circ_0045714-,pcDNA-,si-circ_0045714- and si-NC-transfected ATDC5 cells were inoculated onto the 6-well plates.After 12-hour culture,the apoptosis rate of chondrocytes in each group was detected by using flow cytometry.⑤The culture medium of chondrocytes was collected from each group,and the levels of tumor necrosis factor-α(TNF-α)and interleukin 6(IL-6)in the supernatant were detected.⑥The total RNA was extracted from chondrocytes in each group for synthesizing the cDNA by reverse transcription,followed by amplification for calculating the expression level of circ_0045714 in chondrocytes.Results:①The survival rate of chondrocytes was lower in IL-1β group compared to normal group(LSD-t=15.396,P=0.000),and lower compared with that of IL-1β+50 μg/mL TTS group,IL-1β+100 μg/mL TTS group,IL-1β+200 μg/mL TTS group and IL-1β+400 μg/mL TTS group(LSD-t=4.879,P=0.047; LSD-t=7.686,P=0.001; LSD-t=10.657,P=0.000; LSD-t=10.073,P=0.000); while the differences between IL-1β group and IL-1β+25 μg/mL TTS group,and between IL-1β+200 μg/mL TTS group and IL-1β+400 μg/mL TTS group were not statistically significant(LSD-t=1.555,P=0.918; LSD-t=0.584,P=0.999).Therefore,the TTS with concentrations of 50,100 and 200 μg/mL were selected for the further experiments and were labeled as L-TTS group,M-TTS group,and H-TTS group,respectively,based on the concentrations.②The proliferation inhibition rate,the apoptosis rate,and the levels of TNF-α and IL-6 in the chondrocytes were higher in IL-1β group compared with those of control group,IL-1β+L-TTS group,IL-1β+M-TTS group,and IL-1β+H-TTS group(the proliferation inhibition rate:LSD-t=55.829,P=0.000; LSD-t=8.879,P=0.001; LSD-t=20.507,P=0.000; LSD-t=30.315,P=0.000; the apoptosis rate:LSD-t=27.508,P=0.000; LSD-t=5.076,P=0.032; LSD-t=11.689,P=0.000; LSD-t=21.284,P=0.000; the level of TNF-α in the chondrocytes:LSD-t=29.990,P=0.000; LSD-t=7.720,P=0.002; LSD-t=17.182,P=0.000; LSD-t=24.615,P=0.000; the level of IL-6 in the chondrocytes:LSD-t=33.441,P=0.000; LSD-t=6.324,P=0.008; LSD-t=15.440,P=0.000; LSD-t=25.096,P=0.000),and they were higher in IL-1β+L-TTS group compared to IL-1β+M-TTS group and IL-1β+H-TTS group(the proliferation inhibition rate:LSD-t=11.627,P=0.000; LSD-t=21.436,P=0.000; the apoptosis rate:LSD-t=6.613,P=0.006; LSD-t=16.209,P=0.000; the level of TNF-α in the chondrocytes:LSD-t=9.463,P=0.000; LSD-t=16.895,P=0.000; the level of IL-6 in the chondrocytes:LSD-t=9.117,P=0.001; LSD-t=18.773,P=0.000),furthermore,they were higher in IL-1β+M-TTS group compared to IL-1β+H-TTS group(LSD-t=9.808,P=0.000; LSD-t=9.595,P=0.000; LSD-t=7.432,P=0.003; LSD-t=9.656,P=0.000).③The expression level of circ_0045714 in the chondrocytes was lower in IL-1β group compared to control group,IL-1β+L-TTS group,IL-1β+M-TTS group,and IL-1β+H-TTS group(LSD-t=43.218,P=0.000; LSD-t=9.487,P=0.000; LSD-t=22.136,P=0.000; LSD-t=34.785,P=0.000),and was lower in IL-1β+L-TTS group compared to IL-1β+M-TTS group and IL-1β+H-TTS group(LSD-t=12.649,P=0.000; LSD-t=25.298,P=0.000),moreover,it was lowly expressed in IL-1β+M-TTS group compared to IL-1β+H-TTS group(LSD-t=12.649,P=0.000).④The expression level of circ_0045714 showed a significant difference in the chondrocytes transfected with pcDNA and pcDNA-circ_0045714(1.00±0.00,4.18±0.14,t=39.342,P=0.000),which indicated the chondrocytes with overexpression of circ_0045714 were successfully constructed.The difference in the expression level of circ_0045714 among the chondrocytes transfected with si-NC and si-circ_0045714 indicated the successful silencing of circ-0045714 in chondrocytes(1.00±0.00,0.28±0.04,t=31.177,P=0.000).⑤The IL-1β+pcDNA-circ_0045714 group showed lower proliferation inhibition rate,apoptosis rate,and TNF-α and IL-6 levels in the chondrocytes compared to IL-1β+pcDNA group(t=16.290,P=0.000; t=11.359,P=0.000; t=11.988,P=0.000; t=12.266,P=0.000).⑥The IL-1β+H-TTS+si-circ_0045714 group exhibited higher proliferation inhibition rate,apoptosis rate,and TNF-α and IL-6 levels in the chondrocytes compared to IL-1β+H-TTS+si-NC group(t=9.586,P=0.001; t=9.120,P=0.001; t=7.069,P=0.002; t=10.548,P=0.001).Conclusion:The TTS can inhibit IL-1β-induced chondrocyte apoptosis and the expression of inflammatory cytokine,which has potential value in treatment of osteoarthritis.It may exert the effects by upregulating the expression of circ_0045714 in the chondrocytes,and the effects are stronger at the concentration of 200 μg/mL.

参考文献/References:

[1] DUAN L,DUAN D,WEI W,et al.MiR-19b-3p attenuates IL-1β induced extracellular matrix degradation and inflammatory injury in chondrocytes by targeting GRK6[J].Mol Cell Biochem,2019,459(1-2):205-214.
[2] 常青,王伟,杨增华,等.黄芩苷对IL-1β诱导大鼠软骨细胞凋亡和炎症反应的抑制作用及相关机制研究[J].中国免疫学杂志,2020,36(21):2603-2607.
[3] PARK Y J,CHO Y R,OH J S,et al.Effects of Tribulus terrestris on monosodium iodoacetate-induced osteoarthritis pain in rats[J].Mol Med Rep,2017,16(4):5303-5311.
[4] 翟凤国,周福波,李厚忠,等.蒺藜皂苷对脑缺血再灌注大鼠炎性细胞因子表达的影响[J].牡丹江医学院学报,2014,35(5):4-6.
[5] 李桂双,卜洁琼,龙剑文.蒺藜皂苷抑制肿瘤坏死因子α和干扰素γ诱导HaCaT细胞的炎症反应[J].中国皮肤性病学杂志,2019,33(2):131-137.
[6] JIANG H,DAI J,ZHANG C,et al.Circ_0045714 alleviates TNF-α-induced chondrocyte injury and extracellular matrix degradation through miR-218-5p/HRAS axis[J].J Bioenerg Biomembr,2021,53(1):97-107.
[7] 齐秀春,陈昕,曹玉净,等.雷公藤多苷通过Wnt/β-catenin缓解IL-1β诱导的软骨细胞损伤[J].中成药,2020,42(11):2890-2896.
[8] LI G,TAN W,FANG Y,et al.circFADS2 protects LPS-treated chondrocytes from apoptosis acting as an interceptor of miR-498/mTOR cross-talking[J].Aging(Albany NY),2019,11(10):3348-3361.
[9] 熊杰,匡浩铭,钟秀远,等.铁包金按摩膏联合刮法对膝骨关节炎模型兔关节软骨IL-1β、TNF-α及A20蛋白表达的影响[J].中医药导报,2022,28(5):42-46.
[10] 张安琪,郑福增,周子朋,等.女贞子多糖对IL-1β诱导的关节软骨细胞损伤和炎症反应的作用[J].中成药,2022,44(5):1447-1453.
[11] 汪珏,郑林峰,徐进,等.骨碎补总黄酮对IL-1β诱导体外软骨细胞损伤的保护作用[J].中国现代应用药学,2021,38(12):1441-1447.
[12] 全惠琳,郇宇,胡学昱.褪黑素对IL-1β/TNF-α诱导终板软骨细胞黏附减少的作用和机制[J].空军军医大学学报,2022,43(5):444-449.
[13] LIU C,REN S,ZHAO S,et al.LncRNA MALAT1/MiR-145 adjusts IL-1β-induced chondrocytes viability and cartilage matrix degradation by regulating ADAMTS5 in human osteoarthritis[J]. Yonsei Med J,2019,60(11):1081-1092.
[14] 王艳霞.蒺藜皂苷通过上调PDCD4表达阻滞肺癌A549细胞周期并诱导细胞凋亡[J].中国药师,2020,23(10):1894-1898.
[15] 朱克春,马萍.蒺藜总皂苷对LPS诱导的巨噬细胞分泌IL-1β、IL-6、TNF-α、IL-2、NO的影响和机制[J].中国免疫学杂志,2021,37(16):1958-1963.
[16] 张素军,冯尚彩.蒺藜皂苷对正常和2型糖尿病大鼠餐后血糖水平的影响[J].实用药物与临床,2012,15(1):1-3.
[17] WANG Y,GUO W,LIU Y,et al.Investigating the protective effect of gross saponins of Tribulus terrestrisfruit against ischemic stroke in rat using metabolomics and network pharmacology[J].Metabolites,2019,9(10):240.
[18] 张爽,李红,梁蕾,等.蒺藜皂苷预适应对大鼠离体心脏缺血再灌注损伤的保护作用[J].吉林大学学报(医学版),2010,36(2):229-232.
[19] TAO S C,HUANG J Y,GAO Y,et al.Small extracellular vesicles in combination with sleep-related circRNA3503:a targeted therapeutic agent with injectable thermosensitive hydrogel to prevent osteoarthritis[J].Bioact Mater,2021,6(12):4455-4469.
[20] LI H,LIU Z,GUO X,et al.Circ_0128846/miR-140-3p/JAK2 network in osteoarthritis development[J].Immunol Invest,2022,51(6):1529-1547.
[21] YUAN X,ZHANG Y,CAI C,et al.Circular RNA circZNF652 is overexpressed in osteoarthritis and positively regulates LPS-induced apoptosis of chondrocytes by upregulating PTEN[J].Autoimmunity,2021,54(7):415-421.
[22] CHEN G,LIU T,YU B,et al.CircRNA-UBE2G1 regulates LPS-induced osteoarthritis through miR-373/HIF-1a axis[J].Cell Cycle,2020,19(13):1696-1705.
[23] SHEN S,WU Y,CHEN J,et al.CircSERPINE2 protects against osteoarthritis by targeting miR-1271 and ETS-related gene[J].Ann Rheum Dis,2019,78(6):826-836.
[24] NI J L,DANG X Q,SHI Z B.CircPSM3 inhibits the proliferation and differentiation of OA chondrocytes by targeting miRNA-296-5p[J].Eur Rev Med Pharmacol Sci,2020,24(7):3467-3475.
[25] DING R,ZHOU J,XU J,et al.Circ_0045714/miR-331-3p interaction affects IL-1β-evoked human articular chondrocyte injury through regulating PIK3R3 in a ceRNA regulatory cascade[J].J Orthop Surg Res,2021,16(1):595.
[26] LI B F,ZHANG Y,XIAO J,et al.Hsa_circ_0045714 regulates chondrocyte proliferation,apoptosis and extracellular matrix synthesis by promoting the expression of miR-193b target gene IGF1R[J].Hum Cell,2017,30(4):311-318.

相似文献/References:

[1]孟维娜,明立功,王新德,等.关节镜下清理联合腓骨近1/3段截骨治疗膝骨关节炎[J].中医正骨,2015,27(11):40.
[2]明立功,孟维娜,王新德,等.腓骨近端截骨治疗内侧间室膝骨关节炎的近期疗效观察[J].中医正骨,2015,27(10):25.
[3]张杰,王人彦,张玉柱.膝骨关节炎的治疗进展[J].中医正骨,2015,27(10):68.
[4]梁朝,蔡静怡,闫立,等.针刀疗法改善膝骨关节炎早期疼痛症状的疗效评价[J].中医正骨,2015,27(09):9.
　LIANG Zhao,CAI Jingyi,YAN Li,et al.Evaluation of the curative effect of needle-knife therapy for relieving knee pain in patients with early knee osteoarthritis[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2015,27(1):9.
[5]王建武,党建军,李强,等.四联疗法治疗膝骨关节炎[J].中医正骨,2015,27(08):44.
[6]刘红娟,郭会利,郭树农.云克联合中药治疗膝骨关节炎的护理[J].中医正骨,2015,27(08):75.
[7]陈卫衡.探索建立系统的膝骨关节炎中医临床科研范式 和理论体系[J].中医正骨,2015,27(07):1.
[8]郑春松,叶蕻芝,李西海,等.透骨消痛胶囊中补肾柔肝药和活血祛风药治疗 骨关节炎作用方式的计算机模拟比较[J].中医正骨,2015,27(07):6.
　ZHENG Chunsong,YE Hongzhi,LI Xihai,et al.Comparison of the mode of action of Bushen Rougan(补肾柔肝)drugs versus Huoxue Qufeng(活血祛风)drugs contained in Tougu Xiaotong Jiaonang(透骨消痛胶囊)for the treatment of osteoarthritis:A computer simulation study[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2015,27(1):6.
[9]帅波,沈霖,杨艳萍,等.加味青娥丸治疗膝骨关节炎的作用机制研究[J].中医正骨,2015,27(07):15.
　SHUAI Bo,SHEN Lin,YANG Yanping,et al.Study on the mechanism of action of Jiawei Qing'e Wan(加味青娥丸)for the treatment of knee osteoarthritis[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2015,27(1):15.
[10]梅其杰,袁长深,段戡,等.壮药骨痹方烫熨联合运动疗法治疗膝骨关节炎的临床研究[J].中医正骨,2015,27(07):27.
　MEI Qijie,YUAN Changshen,DUAN Kan,et al.Clinical study of the curative effect of hot compressing and rubbing with packet of Gubi Fang(骨痹方)combined with exercise therapy in the treatment of knee osteoarthritis[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2015,27(1):27.
[11]郑春松,叶蕻芝,李西海,等.独活寄生汤含药血清对白细胞介素1β诱导的 退变关节软骨细胞中基质金属蛋白酶 和环氧化酶2表达的影响[J].中医正骨,2015,27(12):1.
　ZHENG Chunsong,YE Hongzhi,LI Xihai,et al.Impact of Duhuo Jisheng Tang(独活寄生汤)medicated serum on expression of matrix metalloproteinase and cyclooxygenase 2 in degenerative articular chondrocytes induced by interleukin-1 beta[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2015,27(1):1.
[12]林木南,林艳红,刘献祥,等.温热疏密波对膝骨关节炎模型大鼠软骨细胞 RAS/丝裂原活化蛋白激酶信号通路的影响[J].中医正骨,2016,28(02):1.
　LIN Munan,LIN Yanhong,LIU Xianxiang,et al.Effect of warm sparse-dense wave on RAS/mitogen-activated protein kinase signaling pathway in chondrocytes of knee osteoarthritis rat models[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2016,28(1):1.
[13]原晓强,金王东,周云婧,等.纯化血小板对大鼠软骨细胞增殖及膝骨关节炎大鼠软骨修复的作用研究[J].中医正骨,2016,28(12):6.
　YUAN Xiaoqiang,JIN Wangdong,ZHOU Yunjing,et al.Effect of purified platelet rich plasma on chondrocyte proliferation in rats and cartilage repair in rats with knee osteoarthritis:an experimental study[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2016,28(1):6.
[14]刘启明,罗统富,董黎强,等.植物雌激素类中药延缓关节软骨细胞退变的研究进展[J].中医正骨,2017,29(05):27.
[15]叶锦霞,付长龙,林洁,等.透骨消痛胶囊对毒胡萝卜素诱导的内质网应激PEKR信号通路介导的大鼠体外培养关节软骨细胞凋亡的影响[J].中医正骨,2017,29(06):1.
　YE Jinxia,FU Changlong,LIN Jie,et al.Effect of Tougu Xiaotong Jiaonang(透骨消痛胶囊)on apoptosis mediated by endoplasmic reticulum stress(PEKR signaling pathway)and induced by thapsigargin in rat's articular chondrocytes cultured in vitro[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2017,29(1):1.
[16]何晓娟,林平冬,马玉环,等.独活寄生汤含药血清抑制白细胞介素1β诱导的软骨细胞炎症反应的作用机制研究[J].中医正骨,2017,29(08):1.
　HE Xiaojuan,LIN Pingdong,MA Yuhuan,et al.Study on mechanism of action of Duhuo Jisheng Tang(独活寄生汤)medicated serum in inhibiting inflammatory reaction induced by interleukin-1 beta in chondrocytes[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2017,29(1):1.
[17]郑文伟,何晓娟,贾良良,等.跳骨片含药血清抑制脂多糖诱导的软骨细胞炎症反应的作用机制研究[J].中医正骨,2017,29(08):8.
　ZHENG Wenwei,HE Xiaojuan,JIA Liangliang,et al.Study on mechanism of action of Tiaogu Pian(跳骨片)medicated serum in inhibiting inflammatory reaction induced by lipopolysaccharides in chondrocytes[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2017,29(1):8.
[18]陈世宣,刘益杰,胡笑燊,等.基于瞬时受体电位香草素亚家族4信号通路探讨牛蒡子苷元对体外培养软骨细胞增殖及Ⅱ型胶原蛋白和软骨蛋白聚糖表达的影响[J].中医正骨,2017,29(10):13.
　CHEN Shixuan,LIU Yijie,HU Xiaoshen,et al.An experimental study of effect of arctigenin on proliferation of chondrocyte cultured in vitro and expression of typeⅡcollagen protein and aggrecan based on the transient receptor potential vanilloid 4 signaling pathway[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2017,29(1):13.
[19]陈俊,吴广文,许惠凤,等.独活寄生汤含药血清对大鼠退变软骨细胞蛋白激酶 R样内质网激酶/免疫球蛋白结合蛋白信号通路的影响[J].中医正骨,2018,30(08):1.
　CHEN Jun,WU Guangwen,XU Huifeng,et al.Effect of Duhuo Jisheng Tang(独活寄生汤)medicated serum on protein kinase R-like endoplasmic reticulum kinase/immunoglobulin-binding protein signaling pathway in degenerated chondrocytes of rats[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2018,30(1):1.
[20]许丽梅,李慧,许云腾,等.基于NF-κB信号通路探讨独活寄生汤抑制脂多糖诱导的软骨细胞炎症反应的作用机制[J].中医正骨,2019,31(07):9.
　XU Limei,LI Hui,XU Yunteng,et al.An experimental study of mechanism of action of Duhuo Jisheng Tang(独活寄生汤)in inhibiting inflammatory reaction induced by lipopolysaccharide in chondrocytes based on NF-κB signaling pathway[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2019,31(1):9.

备注/Memo

备注/Memo:
基金项目:保定市科技计划项目(2041ZF019)
更新日期/Last Update: 1900-01-01