[1]丁平,张岱阳△,李贺伟,等.积雪草苷对膝骨关节炎大鼠软骨损伤的影响及机制研究[J].中国中医骨伤科杂志,2022,30(09):7-13+19.
　DING Ping,ZHANG Daiyang,LI Hewei,et al.Impacts and Mechanism Study of Asiaticoside on Cartilage Damage and OPG/RANK/RANKL Signaling Pathway in Rats with Knee Osteoarthritis[J].Chinese Journal of Traditional Medical Traumatology & Orthopedics,2022,30(09):7-13+19.
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积雪草苷对膝骨关节炎大鼠软骨损伤的影响及机制研究()
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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09期
页码:
7-13+19
栏目:
实验研究
出版日期:
2022-09-15

文章信息/Info

Title:
Impacts and Mechanism Study of Asiaticoside on Cartilage Damage and OPG/RANK/RANKL Signaling Pathway in Rats with Knee Osteoarthritis
文章编号:
1005-0205(2022)09-0007-07
作者:
丁平1; 张岱阳1△; 李贺伟2; 吴燕茹2; 张光恒3
1恩施土家族苗族自治州中心医院脊柱外科(湖北 恩施,445000)
2华中科技大学同济医学院附属梨园医院
3华中科技大学同济医学院附属梨园医院中心实验室
Author(s):
DING Ping1; ZHANG Daiyang1△; LI Hewei2; WU Yanru2; ZHANG Guangheng3
1Department of Spinal Surgery,Central Hospital of Enshi Tujia and Miao Autonomous Prefecture,Enshi 445000,Hubei China; 2Liyuan Hospital Affiliated to Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430077,China; 3Central Laboratory of Liyuan Hospital Affiliated to Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430077,China.
关键词:
积雪草苷;  膝骨关节炎;  骨保护素;  核因子κB受体活化因子;  核因子κB受体活化因子配体
Keywords:
asiaticoside;  knee osteoarthritis;  osteoprotegerin;  receptor activator of nuclear factor κB;  receptor activator of nuclear factor κB ligand
分类号:
R-33
文献标志码:
A
摘要:
目的:探讨积雪草苷(ASS)对膝骨关节炎(KOA)大鼠软骨损伤和骨保护素(OPG)/核因子κB受体活化因子(RANK)/RANK配体(RANKL)信号通路的影响。方法:采用前交叉韧带切断术(ACLT)加内侧半月板切除术(MMx)诱导建立膝骨关节炎大鼠模型,建模后分为KOA组、ASS低剂量组(ASS-L组,16 mg/kg)、ASS高剂量组(ASS-H组,32 mg/kg)、ASS+si-NC组(32 mg/kg ASS+100 ng LV-si-NC载体)、ASS+si-OPG组(32 mg/kg ASS+100 ng LV-si-OPG载体),另设假手术(Sham)组,每组12只。给予相应的药物干预2周后,通过机械缩爪阈值(PWT)和热缩爪潜伏期(PWL)评估大鼠疼痛行为,酶联免疫吸附(ELISA)法检测血清炎性细胞因子白细胞介素(IL)-1β、IL-6和肿瘤坏死因子-α(TNF-α)水平,苏木精-伊红(HE)和番红O/固绿染色观察关节软骨组织病理学变化,实时荧光定量PCR(RT-qPCR)检测软骨组织基质金属蛋白酶(MMP-13)、Ⅱ型胶原蛋白(COL2A1)、聚集蛋白聚糖(Aggrecan)mRNA表达,蛋白质印迹(Western Blot)检测软骨组织OPG、RANK、RANKL蛋白表达。结果:与Sham组相比,KOA组大鼠的PWT和PWL值、COL2A1 mRNA、Aggrecan mRNA和OPG蛋白水平显著降低,血清IL-1β、IL-6和TNF-α水平、软骨组织Mankin评分和OARSI评分、MMP-13 mRNA和RANK、RANKL蛋白水平显著升高,差异有统计学意义(P<0.05); 与KOA组相比,ASS-L组和ASS-H组大鼠的PWT和PWL值、COL2A1 mRNA、Aggrecan mRNA和OPG蛋白水平显著升高,血清IL-1β、IL-6和TNF-α水平、Mankin评分和OARSI评分、MMP-13 mRNA和RANK、RANKL蛋白水平显著降低,差异有统计学意义(P<0.05); 抑制OPG可明显逆转积雪草苷对膝骨关节炎大鼠软骨损伤的保护作用。结论:积雪草苷可减少促炎细胞因子的释放,抑制MMP-13表达和增加Ⅱ型胶原沉积,抑制关节软骨退变,并可能通过调节OPG/RANK/RANKL信号通路,发挥抗膝骨关节炎作用。
Abstract:
Objective:To investigate the impacts of asiaticoside(ASS)on cartilage damage and osteoprotegerin(OPG)/receptor activator of nuclear factor κB(RANK)/RANK ligand(RANKL)signaling pathway in knee osteoarthritis(KOA)rats.Methods:KOA rat model was established by induction of anterior cruciate ligament amputation(ACLT)and medial meniscectomy(MMx).After modeling,they were grouped into KOA group,ASS low-dose group(ASS-L group,16 mg/kg),ASS high-dose group(ASS-H group,32 mg/kg),ASS+si-NC group(32 mg/kg ASS+100 ng LV-si-NC carrier)and ASS+si-OPG group(32 mg/kg ASS+100 ng LV-si-OPG carrier),and another sham operation(Sham)group was set,with 12 animals in each group.After 2 weeks of corresponding drug intervention,the pain behavior of rats was evaluated by mechanical paw withdrawal threshold(PWT)and thermal paw withdrawal latency(PWL).The levels of serum inflammatory cytokines interleukin(IL)-1β,IL-6 and tumor necrosis factor-α(TNF-α)were measured by enzyme-linked immunosorbent assay(ELISA).The histopathological changes of articular cartilage were observed by hematoxylin-eosin(HE)and safranin O/fast green staining.The mRNA expression of matrix metalloproteinase 13(MMP-13),collagen type Ⅱ(COL2A1)and aggrecan(Aggrecan)in cartilage tissue was measured by real-time quantitative PCR(RT-qPCR),and the protein expression of OPG,RANK and RANKL in cartilage tissue was measured by Western Blot.Results:Compared with the Sham group,the PWT and PWL values,COL2A1 mRNA,Aggrecan mRNA and OPG protein levels were significantly decreased in the KOA group; the serum IL-1β,IL-6 and TNF-α levels,cartilage tissue Mankin scores and OARSI scores,MMP-13 mRNA and RANK,RANKL protein levels were significantly increased(P<0.05).Compared with the KOA group,the PWT and PWL values,COL2A1 mRNA,Aggrecan mRNA and OPG protein levels of the ASS-L group and ASS-H group were significantly increased; the serum IL-1β,IL-6 and TNF-α levels,Mankin scores and OARSI scores,MMP-13 mRNA and RANK,RANKL protein levels were significantly decreased(P<0.05).Inhibition of OPG was able to significantly reverse the protective efficacy of ASS on cartilage injury in KOA rats(P<0.05).Conclusion:ASS can reduce the release of pro-inflammatory cytokines,inhibit the expression of MMP-13,increase the deposition of type Ⅱ collagen and inhibit the degeneration of articular cartilage,which may play an anti-KOA effect by regulating the OPG/RANK/RANKL signaling pathway.

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备注/Memo

备注/Memo:
基金项目:武汉市卫生健康委员会资助项目(武卫通〔2019〕87号)
△通信作者 E-mail:ei68pz@163.com
更新日期/Last Update: 2022-09-10