[1]吕先俊 常利民 陈同林 刘宝戈.夏枯草黄酮对骨关节炎大鼠软骨细胞凋亡的影响及作用机制研究[J].中国中医骨伤科杂志,2022,30(03):1-6.
　LYU Xianjun CHANG Limin CHEN Tonglin LIU Baoge.Study of Efficacy of Prunella Vulgaris Flavonoids on Chondrocyte Apoptosis in Osteoarthritis Rats and Its Action Mechanism[J].Chinese Journal of Traditional Medical Traumatology & Orthopedics,2022,30(03):1-6.
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夏枯草黄酮对骨关节炎大鼠软骨细胞凋亡的影响及作用机制研究()
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《中国中医骨伤科杂志》[ISSN:1005-0205/CN:42-1340/R]

卷:
第30卷
期数:
2022年03期
页码:
1-6
栏目:
实验研究
出版日期:
2022-03-15

文章信息/Info

Title:
Study of Efficacy of Prunella Vulgaris Flavonoids on Chondrocyte Apoptosis in Osteoarthritis Rats and Its Action Mechanism
文章编号:
1005-0205(2022)03-0001-06
作者:
吕先俊1 常利民1 陈同林1 刘宝戈2
1北京朝阳急诊抢救中心创伤骨科(北京,100020)2首都医科大学附属北京天坛医院骨科
Author(s):
LYU Xianjun1 CHANG Limin1 CHEN Tonglin1 LIU Baoge2
1Department of Trauma Orthopedics, Beijing Chaoyang Emergency Rescue Center, Beijing 100020, China; 2Department of Orthopedics, Beijing Tiantan Hospital Affiliated to Capital Medical University, Beijing 100070, China.
关键词:
夏枯草黄酮;  骨关节炎;  大鼠;  软骨细胞;  凋亡
Keywords:
prunella vulgaris flavonoids;  osteoarthritis;  rat;  chondrocyte;  apoptosis
分类号:
R-33
文献标志码:
A
摘要:
目的:探讨夏枯草黄酮对骨关节炎(OA)大鼠软骨细胞凋亡及Ras同源基因家族成员A(RhoA)/Rho相关卷曲螺旋蛋白激酶(ROCK)通路的影响。方法:通过切断大鼠前交叉副韧带、内侧副韧带,摘除内侧半月板构建OA大鼠模型,将造模成功大鼠随机分为OA组、夏枯草黄酮低、中、高剂量组和RhoA通路抑制剂(Y-27632)组; 另取12只大鼠仅剪开皮肤暴露关节腔,作为假手术组; 假手术组和OA组大鼠均给予生理盐水灌胃,夏枯草黄酮低、中、高剂量组分别给予6.125,12.250,24.500 mg/kg夏枯草黄酮灌胃,Y-27632组给5 mg/kg的Y-27632灌胃; 1次/d,连续8周。末次给药后24 h,检测大鼠压痛阈值、热痛阈值,酶联免疫吸附法检测大鼠血清肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)水平,苏木精-伊红(HE)染色法观察大鼠关节软骨形态,一步法原位末端标记(TUNEL)法检测大鼠软骨细胞凋亡,蛋白免疫印迹法检测大鼠软骨组织凋亡蛋白半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)及RhoA/ROCK通路蛋白的表达。结果:假手术组大鼠关节软骨结构清晰,排列规整,表层光滑无缺损,各层细胞分布均匀; OA组大鼠关节软骨表层粗糙且不完整,纤维化明显,各层细胞排列紊乱; 夏枯草黄酮各剂量组、Y-27632组大鼠表层结构相对正常,但部分区域仍存在细胞紊乱现象; 夏枯草黄酮高剂量组与Y-27632组两组间大鼠骨关节软骨形态差异不明显。与假手术组比较,OA组大鼠压痛阈值及热痛阈值显著降低,Mankin’s评分、血清TNF-α及IL-1β水平、软骨细胞凋亡数、软骨组织Caspase-3、RhoA、ROCK蛋白表达显著升高(P<0.05); 与OA组比较,夏枯草黄酮低、中、高剂量组压痛阈值及热痛阈值显著升高,Mankin’s评分、血清TNF-α及IL-1β水平、软骨细胞凋亡数、软骨组织Caspase-3、RhoA、ROCK蛋白表达显著降低,且呈现夏枯草黄酮剂量依赖性(P<0.05); 夏枯草黄酮高剂量组和Y-27632组上述指标差异无统计学意义(P>0.05)。结论:夏枯草黄酮通过抑制OA大鼠软骨细胞凋亡改善大鼠关节软骨损伤,可能与抑制RhoA/ROCK信号通路有关。
Abstract:
Objective:To investigate the efficacy of prunella vulgaris flavonoids on chondrocyte apoptosis and rat’s homolog gene family member A(RhoA)/Rho-associated coiled-coil containing protein kinase(ROCK)pathway in osteoarthritis(OA)rats.Methods:The OA rat model was constructed by cutting off the rat’s anterior cruciate collateral ligament and medial collateral ligament and removing the medial meniscus.These model rats were randomly divided into OA group, prunella vulgaris flavonoids low, medium and high dose groups and RhoA pathway inhibitor(Y-27632)group,other 12 rats were cut open only to expose the joint cavity as the sham operation group,rats in sham operation group and OA group were given normal saline by gavage,prunella vulgaris flavonoids low, medium and high dose groups were given 6.125, 12.250 and 24.500 mg/kg prunella flavone by gavage respectively, and Y-27632 group was given 5 mg/kg Y-27632 by gavage once a day for 8 weeks.The tenderness threshold and heat pain threshold of rats were measured; enzyme-linked immunosorbent assay was used to detect the levels of serum tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)in rats,hematoxylin-eosin(HE)staining method was used to observe the morphology of rat articular cartilage; one-step in situ end labeling(TUNEL)method was used to detect rat chondrocyte apoptosis,Western Blot was used to detect the expression of apoptotic protein Caspase-3 and RhoA/ROCK pathway protein in rat cartilage tissue 24 h after the last administration.Results:The articular cartilage structure of rats in the sham operation group was clear and regular,the surface was smooth without defects, and the cells of each layer were evenly distributed.The surface of the articular cartilage of rats in the OA group was rough and incomplete, with obvious fibrosis, and the cell arrangement of each layer was disordered,the surface structure of rats in the prunella vulgaris flavonoids each dose group and Y-27632 group was relatively normal, but there were still cell disorders in some areas,there was no significant difference in the morphology of the bone articular cartilage between the high-dose prunella vulgaris flavonoids group and the Y-27632 group.Compared with the sham operation group, the tenderness threshold and heat pain threshold of rats in the OA group were significantly reduced, and the Mankin’s score, serum TNF-α, IL-1β levels, chondrocyte apoptosis number, cartilage tissue Caspase-3, RhoA and ROCK protein expression were significantly increased(P<0.05).Compared with the OA group, the tenderness threshold and heat pain threshold of rats in the prunella vulgaris flavonoids low, medium, and high dose groups were significantly increased, and the Mankin’s score, serum TNF-α, IL-1β levels, chondrocyte apoptosis number, cartilage tissue Caspase-3, RhoA and ROCK protein expression were significantly reduced, and showed dose-dependent of the prunella vulgaris flavonoids(P<0.05),there was no significant difference in the above indexes between prunella vulgaris flavonoids high dose group and Y-27632 group(P>0.05).Conclusion:Prunella vulgaris flavonoids can improve rat articular cartilage damage by inhibiting chondrocyte apoptosis in osteoarthritis rats, which may be related to the inhibition of RhoA/ROCK signaling pathway.

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备注/Memo

备注/Memo:
基金项目:北京市科学技术委员会科研计划项目(D191100000417005)
更新日期/Last Update: 1900-01-01