[1]刘坦,刘德仁,李振珲,等.当归-牛膝药对调控脑和肌肉芳烃受体核转运样蛋白1/时钟节律调节蛋白信号通路对膝骨关节炎大鼠软骨细胞凋亡的影响[J].中国中医骨伤科杂志,2024,32(12):1-9.[doi:10.20085/j.cnki.issn1005-0205.241201 ]
　LIU Tan,LIU Deren,LI Zhenhui,et al.Effects of Couplet Medicinals of Angelica Sinensis and Achyranthes Bidentata Modulation of Bmal1/Clock Signaling Pathway on Cartilage Cells Apoptosis in Rats with Knee Osteoarthritis[J].Chinese Journal of Traditional Medical Traumatology & Orthopedics,2024,32(12):1-9.[doi:10.20085/j.cnki.issn1005-0205.241201 ]
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当归-牛膝药对调控脑和肌肉芳烃受体核转运样蛋白1/时钟节律调节蛋白信号通路对膝骨关节炎大鼠软骨细胞凋亡的影响()
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《中国中医骨伤科杂志》[ISSN:1005-0205/CN:42-1340/R]

卷:
第32卷
期数:
2024年12期
页码:
1-9
栏目:
实验研究
出版日期:
2024-12-15

文章信息/Info

Title:
Effects of Couplet Medicinals of Angelica Sinensis and Achyranthes Bidentata Modulation of Bmal1/Clock Signaling Pathway on Cartilage Cells Apoptosis in Rats with Knee Osteoarthritis
文章编号:
1005-0205(2024)12-0001-09
作者:
刘坦1; 刘德仁1; 李振珲1; 吴鹏1; 茆军1△
1南京中医药大学附属医院/江苏省中医院(南京,210029)
Author(s):
LIU Tan1; LIU Deren1; LI Zhenhui1; WU Peng1; MAO Jun1△
1Affiliated Hospital of Nanjing University of Chinese Medicine(Jiangsu Province Hospital of Chinese Medicine),Nanjing 210029,China.
关键词:
膝骨关节炎;  当归-牛膝药对;  脑和肌肉芳香烃受体核转运样蛋白1/时钟节律调节蛋白信号通路;  凋亡
Keywords:
knee osteoarthritis;  couplet medicinals of Angelica Sinensis and Achyranthes Bidentata;  brain and muscle ARNT-like 1/clock rhythm regulating protein signaling pathway;  apoptosis
分类号:
R-33
DOI:
10.20085/j.cnki.issn1005-0205.241201
文献标志码:
A
摘要:
目的:探讨当归-牛膝药对调控脑和肌肉芳香烃受体核转运样蛋白1(Bmal1)/ 时钟节律调节蛋白(Clock)信号通路对膝骨关节炎(KOA)大鼠软骨细胞凋亡的影响。方法:采用前交叉韧带横断术(ACLT)构建大鼠膝骨关节炎模型,造模成功后的大鼠随机分为模型组、阳性对照组(塞来昔布18 mg/kg)、当归-牛膝药对低剂量组(1.08 g/kg)、当归-牛膝药对高剂量组(2.16 g/kg)、当归-牛膝+Bmal1抑制剂组(当归-牛膝2.16 g/kg+Bmal1抑制剂SR9009 50 mg/kg),每组7只; 另取7只大鼠作为假手术组。假手术组和模型组大鼠灌胃或腹腔注射生理盐水,其余各组大鼠灌胃或腹腔注射相应药物,1次/d,连续28 d。末次给药后,观察各组大鼠膝关节软骨组织病理改变并进行Mankin评分,检测血清中白细胞介素1β(IL-1β)及IL-6的含量,检测软骨细胞凋亡情况,检测软骨组织中Bmal1蛋白阳性表达,测定软骨组织中B细胞淋巴瘤2(Bcl-2)、Bcl-2相关X蛋白(Bax)、活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved caspase 3)凋亡相关蛋白的表达,软骨组织Bmal1/Clock信号通路相关蛋白及mRNA表达。结果:与假手术组比较,模型组大鼠软骨表面破损、连续性中断,软骨细胞排列紊乱,软骨基质中蛋白聚糖含量降低; 血清中IL-1β、IL-6含量显著升高; 凋亡的软骨细胞显著增多; 软骨组织中Bmal1蛋白阳性表达显著降低,Bcl-2蛋白表达显著降低,Bax、Cleaved caspase 3蛋白表达显著升高,Bmal1、Clock蛋白及mRNA显著升高(P<0.05)。与模型组比较,当归-牛膝药对低、高剂量组和阳性对照组大鼠软骨组织的病理改变均显著改善,上述指标均显著逆转(P<0.05); 抑制Bmal1/Clock信号通路能减轻当归-牛膝药对改善软骨细胞凋亡的作用。结论:当归-牛膝药对可改善膝骨关节炎大鼠软骨细胞凋亡,其作用可能与激活Bmal1/Clock信号通路有关。
Abstract:
Objective:To investigate the effects of couplet medicinals of Angelica Sinensis and Achyranthes Bidentata modulation of brain and muscle ARNT-like 1(Bmal1)/clock rhythm regulating protein(Clock)signaling pathway on cartilage cells apoptosis in rats with knee osteoarthritis(KOA).Methods:Anterior cruciate ligament transection(ACLT)was used to construct a rat KOA model,and the rats after successful modeling were randomly divided into model group,positive control group(celecoxib 18 mg/kg),Angelica Sinensis and Achyranthes Bidentata low-dose group(1.08 g/kg),Angelica Sinensis and Achyranthes Bidentata high-dose group(2.16 g/kg),Angelica Sinensis and Achyranthes Bidentata + Bmal1 inhibitor group(Angelica Sinensis and Achyranthes Bidentata 2.16 g/kg + Bmal1 inhibitor SR9009 50 mg/kg),with 7 rats in each group; another 7 rats were taken as the sham operation group. The rats in the sham-operation group and model group were injected with saline by gavage or intraperitoneal injection,and the rats in the remaining groups were injected with the corresponding drugs by gavage or intraperitoneal injection once a day for 28 d. After the last administration,the histopathological changes in the cartilage of the knee joints of rats in each group were observed and Mankin score was performed,the levels of interleukin 1β(IL-1β)and IL-6 in serum were detected,the chondrocyte apoptosis was detected,the positive expression of Bmal1 protein in cartilage tissue was detected,and the expression of B-cell lymphoma 2(Bcl-2),Bcl-2-associated X protein(Bax),and the expression of cleaved cysteinyl aspartate specific proteinase 3( Cleaved caspase 3)apoptosis-related protein expression,and cartilage tissue Bmal1/Clock signaling pathway-related protein and mRNA expression.Results:Compared with the sham operation group,rats in the model group had broken cartilage surfaces,interrupted continuity,disorganized chondrocyte arrangement,and reduced proteoglycan content in cartilage matrix; the serum levels of IL-1β and IL-6 were significantly elevated; the apoptotic chondrocytes were significantly elevated; the percentage of Bmal1 protein-positive cells in the cartilage tissues was significantly lower,the expression of Bcl-2 protein was significantly lower,and the expressions of Bax,Cleaved caspase 3 protein were significantly elevated,as well as Bmal1,Clock protein and mRNA were significantly elevated(P<0.05). Compared with the model group,the couplet medicinals of Angelica Sinensis and Achyranthes Bidentata pairs of low- and high-dose groups and the positive control group significantly improved the pathological changes in the cartilage tissue of rats,and all of the above indexes were significantly reversed(P<0.05); the inhibition of the Bmal1/Clock signaling pathway could attenuate the effects of the couplet medicinals of Angelica Sinensis and Achyranthes Bidentata on the improvement of apoptosis of chondrocytes.Conclusion:The couplet medicinals of Angelica Sinensis and Achyranthes Bidentata ameliorated cartilage cells apoptosis in KOA rats,and its effect may be related to the activation of the Bmal1/Clock signaling pathway.

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备注/Memo

备注/Memo:
基金项目:国家自然科学基金项目(82205143)
江苏省中医院第三批高峰学术人才项目(y2021rc20)
江苏省中医院优秀青年博士培养专项(2023QB0122)
△通信作者 E-mail:maojun@njucm.edu.cn
更新日期/Last Update: 2024-12-05