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| Study on the mechanism of medicinal bath of Yaoyi Chumen Decoction in the treatment of rats with knee osteoarthritis based on p38MAPK pathway |
| LI Tong1 WANG Yijin1 TANG Yizhou1 MA Yan2 FU Biaofang3 YAN Guoyue4 |
1. College of Yao Medicine, Guangxi University of Chinese Medicine, Guangxi Zhuang Autonomous Region, Nanning 530001, China;
2.Guangxi International Zhuang Medicine Hospital, Guangxi Zhuang Autonomous Region, Nanning 530001, China;
3.College of Basic Medicine, Guangxi University of Chinese Medicine, Guangxi Zhuang Autonomous Region, Nanning 530001, China;
4.Guangxi Talent Highland for Zhuang and Yao Medicine and Combination and Medical Care and Elderly Care, Guangxi University of Chinese Medicine, Guangxi Zhuang Autonomous Region, Nanning 530001, China |
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Abstract Objective To investigate the effect and mechanism of medicinal bath of Yaoyi Chumen Decoction in the treatment of knee osteoarthritis (KOA) model rats based on p38 mitogen-activated protein kinase (p38MAPK) pathway. Methods Ten of 60 8-week-old male SPF SD rats (weight 200-220 g) were randomly selected as blank group, 0.1 ml of physiological saline was injected into the right knee cavity of the blank group on the 1st and 4th days of the experiment. The remaining rats were injected with a mixed emulsion of 0.1 ml type Ⅱ collagenic proteinase and complete Freund’s adjuvant (mass concentration was 0.5 mg/ml). On the 7th day, the modeling results were evaluated according to the knee joint conditions of the rats, 50 rats successfully modeled were divided into model group, Diclofenac Sodium group, and Chumen Decoction high, medium, low dose groups with ten rats in each group according to random number table method. On the 8th day of the experiment, the Chumen Decoction high, medium, low dose groups were given medicinal bath intervention (the concentration was 49.00, 24.50, 12.25 g/L, respectively), the model group was given warm water bath, the Diclofenac Sodium group was given 0.01 g of 1% Diclofenac Sodium Gel at the modeling site, and the blank group was fed routinely, the intervention lasted for four weeks in each group. The general condition of each group was observed during the intervention. After intervention, the anatomy of knee joint in each group was observed, the diameter of knee joint was measured, and the histological changes of knee joint cartilage were observed by hematoxylin-eosin (HE) staining. The serum levels of interleukin-1β(IL-1β), IL-6, and IL-10 were detected by enzyme-linked immunosorbent assay. Immunohistochemical staining was used to detect the location and expression levels of p38MAPK, phosphorylated p38MAPK (p-p38MAPK), and matrix metalloproteinase-13 (MMP-13) in cartilage tissue of each group. Results During the drug intervention, the general condition of the blank group was normal, the model group limp during forced exercise with knee flexion and extension dysfunction, the flexion and extension function of knee joint and gait of Chumen Decoction dose groups were gradually recovered, and the lameness of both feet was reduced. After the intervention, the knee joint anatomy of the blank group showed smooth and translucent synovium, transparent, and smooth cartilage, and complete articular surface, HE staining showed that the chondrocytes were arranged regularly and clearly. The knee joint anatomy of the model group showed synovial thickening and erosion, cartilage surface roughness, and wear, and gray color, HE staining showed that cartilage matrix was reduced or even disappeared, chondrocytes were reduced in large numbers, and the shape was irregular. The knee joint anatomy and HE staining showed that the knee joint cartilage of Chumen Decoction dose groups were improved to varying degrees. The knee diameter of model group was higher than that of blank group, the expressions of serum IL-1β, IL-6, and cartilage p38MAPK, p-p38MAPK, MMP-13 were higher than those of blank group, and serum IL-10 was lower than that of control group (P＜0.05). The knee diameter of each Chumen Decoction dose groups were lower than that of model group, the serum IL-10 was higher than that of model group, and the expressions of cartilage p-p38MAPK and MMP-13 were lower than those of model group (P＜0.05). Serum IL-1β, IL-6, and cartilage p38 MAPK in Chumen Decoction high and medium dose groups were lower than those in model group and Chumen Decoction low dose group, and serum IL-10 was higher than that in Chumen Decoction low dose group (P＜0.05). The knee diameter of Chumen Decoction high dose group was lower than those of Chumen Decoction low and medium dose groups, the expressions of cartilage p-p38MAPK and MMP-13 were lower than those of Chumen Decoction low dose group, the serum IL-10 was higher than that of Chumen Decoction low dose group, and the expression of cartilage MMP-13 was lower than that of Chumen Decoction medium group (P＜0.05). Conclusion Yaoyi Chumen Decoction may reduce the inflammatory response and improve cartilage degeneration in rats with KOA through targeted inhibition of p38MAPK pathway expression.
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