DataSheet_1_Salvianolic Acid A Has Anti-Osteoarthritis Effect In Vitro and In Vivo.zip

Osteoarthritis (OA) is a degenerative disease found in middle-aged and elderly people, which seriously affects their quality of life. The anti-inflammatory and anti-apoptosis pharmacological effects of salvianolic acid A (SAA) have been shown in many studies. In this study, we intended to explore the anti-inflammatory and anti-apoptotic effects of SAA in OA. We evaluated the expression of pro-inflammatory mediators and cartilage matrix catabolic enzymes in chondrocytes by ELISA, Griess reaction, immunofluorescence, and Western blot, which includes nitric oxide (NO), tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), prostaglandin E2 (PGE2), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), MMPs (MMP-3, MMP-13), and ADAMTS-5. Bax, Bcl-2, and cleaved caspase-3 were also measured by Western blot methods. The results of this experiment in vitro showed that SAA not only inhibited the production of inflammatory mediators induced by IL-1β and the loss of cartilage matrix but also reduced the apoptosis of mouse chondrocytes induced by IL-1β. According to the results of immunofluorescence and Western blot, SAA inhibited the activation of the NF-κB pathway and MAPK pathway. The results of these in vitro experiments revealed for the first time that SAA down-regulated the production of inflammatory mediators and inhibited the apoptosis of mouse chondrocytes and the degradation of extracellular matrix (ECM), which may be attributed to the inhibition of the activation of NF-κB and MAPK signaling pathways. In the in vivo experiments, 45 mice were randomly divided among three groups (the sham group, OA group, and OA + SAA group). The results of animal experiments showed that SAA treatment for eight consecutive weeks inhibited further deterioration of OA. These results demonstrate that SAA plays an active therapeutic role in the development of OA.

骨关节炎（Osteoarthritis, OA）是一种常见于中老年人群的退行性疾病，严重影响患者的生活质量。丹参酚酸A（salvianolic acid A, SAA）的抗炎、抗凋亡药理作用已被多项研究证实。本研究旨在探讨SAA对OA的抗炎及抗凋亡作用。我们通过酶联免疫吸附测定（ELISA）、格里斯（Griess）反应、免疫荧光及蛋白质印迹（Western blot）实验，检测了软骨细胞中促炎介质与软骨基质分解酶的表达水平，涵盖一氧化氮（nitric oxide, NO）、肿瘤坏死因子-α（tumor necrosis factor-alpha, TNF-α）、白细胞介素-6（interleukin-6, IL-6）、前列腺素E2（prostaglandin E2, PGE2）、诱导型一氧化氮合酶（inducible nitric oxide synthase, iNOS）、环氧化酶-2（cyclooxygenase-2, COX-2）、基质金属蛋白酶（MMPs, MMP-3、MMP-13）及ADAMTS-5。同时通过蛋白质印迹法检测了Bax、Bcl-2及裂解型半胱氨酸天冬氨酸蛋白酶-3（cleaved caspase-3）的表达水平。体外实验结果显示，SAA不仅可抑制IL-1β诱导的促炎介质产生及软骨基质丢失，还能减轻IL-1β诱导的小鼠软骨细胞凋亡。免疫荧光及蛋白质印迹结果表明，SAA可抑制核因子-κB（NF-κB）通路与丝裂原活化蛋白激酶（MAPK）通路的激活。本体外实验结果首次揭示，SAA可下调促炎介质的产生，抑制小鼠软骨细胞凋亡及细胞外基质（extracellular matrix, ECM）降解，其机制可能与阻断NF-κB及MAPK信号通路的激活有关。体内实验中，我们将45只小鼠随机分为三组：假手术组、OA模型组及OA+SAA给药组。动物实验结果显示，连续8周给予SAA治疗可抑制OA的进一步恶化。上述结果表明，SAA在OA的发生发展中具有积极的治疗作用。