河蚌葡聚糖膝关节内注射对豚鼠软骨退变的影响
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上海中医药大学附属曙光医院

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Effects of intra-articular injection of HBP-A on cartilage degeneration in guinea pigs
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Shi’s Center of Orthopedics and Traumatology, Shuguang Hospital Affiliated to Shanghai University of TCM

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    摘要:

    目的:观察关节内注射不同浓度HBP-A对豚鼠膝关节软骨退变及相关促关节软骨分解代谢因子的影响。方法:豚鼠右膝行前交叉韧带切断术,术后右膝关节内分别每周注射1次低、中、高浓度的HBP-A 200μl,连续注射5周。采用SO法观察右胫骨关节软骨组织退变及其损伤分数。各组胫骨关节软骨Runx2、Ihh、ALP、MMP13蛋白表达采用免疫组织化学法检测并进行定量分析。qPCR检测各组股骨关节软骨Ihh、IL-1β、MMP-13和Runx2的mRNA表达。结果:SO法显示模型组软骨明显磨损,软层全部消失或深层断裂,多糖含量减少,细胞出现聚集并且肥大、坏死和排列紊乱,与正常对照组比较损伤评分增加了9倍,差异具有统计学意义(P<0.05)。在给与不同浓度HBP-A关节腔注射后,软骨磨损情况逐渐好转,组织内细胞死亡数降低,蛋白多糖含量丢失减少,OARSI评分也逐渐减少,差异具有统计学意义(P<0.05)。各浓度组胫骨关节软骨Runx2、Ihh、ALP、MMP13蛋白表达明显降低,差异具有统计学意义(P<0.05)。各浓度组股骨关节软骨Ihh、IL-1β、MMP-13和Runx2的mRNA表达同时逐步减小,差异具有统计学意义(P<0.05)。结论:关节腔内注射HBP-A能够减少或延缓豚鼠膝关节软骨退变程度。

    Abstract:

    To observe the effects of intra-articular injection of different concentrations of anodonta glucan HBP-A on knee cartilage degeneration and related factors promoting articular cartilage catabolism in guinea pigs. Methods: Anterior cruciate ligament (ACL) amputation was performed on the right knee joint of guinea pigs. 200μl low, moderate and high concentration of HBP-A were injected into the right knee joint after operation, and then once a week for 5 weeks. The degeneratione the injury score of articular cartilage were measured by OARSI method with Safranine-solid green staining (SO). The protein expressions of Runx2, Ihh, ALP and MMP13 in tibial articular cartilage were detected by immunohistochemical method. qPCR was used to detect the mRNA expression of Ihh, IL-1β, MMP-13 and Runx2 in femoral articular cartilage. Results: SO method showed that the cartilage of the model group was obviously worn, the soft layer disappeared or deeply broken, the content of polysaccharide decreased, and the cells gathered, hypertrophy, necrosis and disorder, and the OARSI score increased 9 times compared with the normal control group, the difference was statistically significant (P < 0.05). After intra-articular injection of different concentrations of HBP-A, cartilage wear was gradually improved, the number of cell death in tissue decreased, the loss of proteoglycan content decreased, and the OARSI score gradually decreased, the difference was statistically significant (P < 0.05). The protein expressions of Runx2, Ihh, ALP and MMP13 in tibial articular cartilage were significantly decreased in all concentration groups (P < 0.05). The mRNA expression of Ihh, IL-1 β, MMP-13 and Runx2 in femoral articular cartilage decreased gradually in all concentration groups, and the difference was statistically significant (P < 0.05). Conclusion: Intra-articular injection of HBP-A can reduce or delay the degree of knee cartilage degeneration in guinea pigs.

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  • 收稿日期:2021-11-28
  • 最后修改日期:2022-02-02
  • 录用日期:2022-06-07
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