Hossain Mohammad Amjad, Alam Md Jahangir, Kim Bumseok, Kang Chang-Won, Kim Jong-Hoon
College of Veterinary Medicine, Biosafety Research Institute, Chonbuk National University, 54596 79 Gobong-ro, Iksan-city, Jeollabuk-Do, Republic of Korea.
College of Veterinary Medicine, Biosafety Research Institute, Chonbuk National University, 54596 79 Gobong-ro, Iksan-city, Jeollabuk-Do, Republic of Korea.
Phytomedicine. 2022 Jun;100:154039. doi: 10.1016/j.phymed.2022.154039. Epub 2022 Mar 11.
Osteoarthritis (OA) is the most common joint complaint resulting in pain, disability, and loss of quality of life. On the other hand, ginsenoside-Rb1 is a plant product derived from ginseng that possesses immune-regulation and anti-inflammatory activities. However, it has been reported that different rout of administration but hydrogel-based Ginsenoside-Rb1 in an OA rabbit model has not been investigated.
The aim of this study was to investigate the potential effects of ginsenoside-Rb1 such as anti-arthritic activity in a rabbit knee OA model via NF- κB, PI3K/Akt, and P38/(MAPK) pathways.
In the current study, rabbit osteoarthritis was induced by hollow trephine on the femur trochlea and the hydrogel-based Ginsenoside-Rb1 sheets were inserted on the rabbit knee to assess the anti-arthritis activity of ginsenoside-Rb1 which is sustained release.
After the hydrogel-based Rb1 sheet insert on the rabbit knee, macroscopic and micro CT was performed for investigation of chondroprotective effect. Matrix metalloproteinases (MMPs) and apoptotic expression were assessed through Immunohistochemistry and RT-PCR assay. In addition, the flow cytometry technique was used for the investigation of intracellular reactive oxygen species (ROS) production and histological changes were examined by HE, safranin O, and Masson trichrome staining method. Furthermore, the NF- κB, PI3K/Akt, and P38/(MAPK) pathways were investigated using Western blot analysis.
Macroscopic and micro CT investigation of hydrogel-Rb1 treatment showed a dose-dependent chondroprotective effect. Immunohistochemistry and RT-PCR revealed that expression of matrix metalloproteinases (MMPs) and apoptotic markers TNF-α, caspase-3, and bax are down-regulated in a dose-dependent fashion following implantation of hydrogel-Rb. Higher levels of intracellular reactive oxygen species (ROS) were observed in the OA group. In histopathological investigation of hydrogel-Rb1 exhibited larger amounts of chondro cells, glycosaminoglycan's, and collagen compared to the defect group. Furthermore, the NF- κB, PI3K/Akt, and P38/(MAPK) pathways were downregulated by hydrogel-Rb1 while the disease model showed upstream. In the meantime, MMP expression level was considerably down-regulated.
Our results indicate the protective effect of ginsenoside-Rb1 against OA pathogenesis through prevention of apoptosis with suppression of ROS production and activation of NF-κB signaling through downregulation of the MAPK and PI3K/Akt signaling pathways.
骨关节炎（OA）是最常见的关节疾病，会导致疼痛、残疾和生活质量下降。另一方面，人参皂苷-Rb1是一种从人参中提取的植物产物，具有免疫调节和抗炎活性。然而，据报道，不同给药途径的基于水凝胶的人参皂苷-Rb1在OA兔模型中的作用尚未得到研究。
本研究旨在通过NF-κB、PI3K/Akt和P38/（MAPK）信号通路，研究人参皂苷-Rb1在兔膝骨关节炎模型中的潜在抗关节炎活性等作用。
在本研究中，通过在股骨滑车处用空心环钻诱导兔骨关节炎，并将基于水凝胶的人参皂苷-Rb1片插入兔膝关节，以评估具有缓释作用的人参皂苷-Rb1的抗关节炎活性。
在兔膝关节插入基于水凝胶的Rb1片后，进行宏观和微观CT检查以研究软骨保护作用。通过免疫组织化学和RT-PCR检测基质金属蛋白酶（MMPs）和凋亡表达。此外，采用流式细胞术检测细胞内活性氧（ROS）的产生，并通过苏木精-伊红（HE）、番红O和Masson三色染色法检查组织学变化。此外，采用蛋白质免疫印迹分析研究NF-κB、PI3K/Akt和P38/（MAPK）信号通路。
水凝胶-Rb1治疗的宏观和微观CT检查显示出剂量依赖性的软骨保护作用。免疫组织化学和RT-PCR显示，植入水凝胶-Rb后，基质金属蛋白酶（MMPs）以及凋亡标志物TNF-α、半胱天冬酶-3和bax的表达呈剂量依赖性下调。OA组观察到较高水平的细胞内活性氧（ROS）。在组织病理学检查中，与缺损组相比，水凝胶-Rb1显示出更多的软骨细胞、糖胺聚糖和胶原蛋白。此外，水凝胶-Rb1可下调NF-κB、PI3K/Akt和P38/（MAPK）信号通路，而疾病模型显示这些信号通路处于上游状态。同时，MMP表达水平显著下调。
我们的结果表明，人参皂苷-Rb1通过抑制ROS产生预防细胞凋亡，并通过下调MAPK和PI3K/Akt信号通路激活NF-κB信号，从而对OA发病机制具有保护作用。
