Green tea polyphenol treatment is chondroprotective, anti-inflammatory and palliative in a mouse post-traumatic osteoarthritis model.
Green tea polyphenol treatment is chondroprotective, anti-inflammatory and palliative in a mouse post-traumatic osteoarthritis model.
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DOI：
10.1186/s13075-014-0508-y
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发表时间：
2014-12-17
影响因子：
4.9
通讯作者：
Sun HB
中科院分区：
文献类型：
--
作者：
Leong DJ;Choudhury M;Hanstein R;Hirsh DM;Kim SJ;Majeska RJ;Schaffler MB;Hardin JA;Spray DC;Goldring MB;Cobelli NJ;Sun HB
表没食子儿茶素3-没食子酸酯（EGCG），一种存在于绿色茶中的多酚，被证明在体外发挥软骨保护作用。在这项研究中，我们使用创伤后骨关节炎（OA）小鼠模型来测试表没食子儿茶素没食子酸酯是否可以减缓OA的进展并减轻OA相关的疼痛。使C57 BL/6小鼠经受内侧半月板（DMM）的手术去稳定化或假手术。从手术当天开始，每天通过腹膜内注射施用EGCG（25 mg/kg）或媒介物对照，持续4或8周。采用番红O染色和国际骨关节炎研究协会（OARSI）评分，以及免疫组织化学分析检测裂解的聚集蛋白聚糖和II型胶原蛋白以及蛋白水解酶基质金属蛋白酶13（MMP-13）和A型解整合素和金属蛋白酶与血小板反应蛋白基序5（ADAMTS 5）的表达来评估OA的严重程度。进行实时PCR以表征关节软骨稳态的关键基因的表达。在实验过程中，触觉敏感性测试（von Frey测试）和旷场试验用于评估与OA相关的疼痛行为，并通过实时PCR测定背根神经节（DRG）中疼痛表达标记物和炎性细胞因子的表达。DMM手术后4周和8周，与溶媒治疗的对照组相比，EGCG治疗的小鼠的软骨显示出较少的番红O损失和软骨侵蚀，以及较低的OARSI评分，这与关节软骨中聚集蛋白聚糖和II型胶原裂解表位的染色减少以及MMP-13和ADAMTS 5的染色减少有关。在接受EGCG治疗的小鼠中，关节软骨也表现出Mmp 1、Mmp 3、Mmp 8、Mmp 13、Adamts 5、白细胞介素1 β（IL 1b）和肿瘤坏死因子α（Tnfa）mRNA水平降低，MMP调节因子Cbp/p300相互作用反式激活因子2的基因表达升高（Cited 2）。与载体对照相比，用EGCG处理的小鼠表现出减少的OA相关疼痛，如由更高的运动行为（即，行进的距离）所指示的。此外，在DRG中的趋化因子受体Ccr 2和促炎细胞因子Illb和Tnfa的表达显著降低至与假手术动物相似的水平。这项研究提供了第一个证据，在OA动物模型中，表没食子儿茶素没食子酸酯显着减缓OA疾病的进展，并发挥姑息作用。本文的在线版本（doi：10.1186/s13075-014-0508-y）包含补充材料，可供授权用户使用。
Epigallocatechin 3-gallate (EGCG), a polyphenol present in green tea, was shown to exert chondroprotective effects in vitro. In this study, we used a posttraumatic osteoarthritis (OA) mouse model to test whether EGCG could slow the progression of OA and relieve OA-associated pain. C57BL/6 mice were subjected to surgical destabilization of the medial meniscus (DMM) or sham surgery. EGCG (25 mg/kg) or vehicle control was administered daily for 4 or 8 weeks by intraperitoneal injection starting on the day of surgery. OA severity was evaluated using Safranin O staining and Osteoarthritis Research Society International (OARSI) scores, as well as by immunohistochemical analysis to detect cleaved aggrecan and type II collagen and expression of proteolytic enzymes matrix metalloproteinase 13 (MMP-13) and A disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS5). Real-time PCR was performed to characterize the expression of genes critical for articular cartilage homeostasis. During the course of the experiments, tactile sensitivity testing (von Frey test) and open-field assays were used to evaluate pain behaviors associated with OA, and expression of pain expression markers and inflammatory cytokines in the dorsal root ganglion (DRG) was determined by real-time PCR. Four and eight weeks after DMM surgery, the cartilage in EGCG-treated mice exhibited less Safranin O loss and cartilage erosion, as well as lower OARSI scores compared to vehicle-treated controls, which was associated with reduced staining for aggrecan and type II collagen cleavage epitopes, and reduced staining for MMP-13 and ADAMTS5 in the articular cartilage. Articular cartilage in the EGCG-treated mice also exhibited reduced levels of Mmp1, Mmp3, Mmp8, Mmp13,Adamts5, interleukin 1 beta (Il1b) and tumor necrosis factor alpha (Tnfa) mRNA and elevated gene expression of the MMP regulator Cbp/p300 interacting transactivator 2 (Cited2). Compared to vehicle controls, mice treated with EGCG exhibited reduced OA-associated pain, as indicated by higher locomotor behavior (that is, distance traveled). Moreover, expression of the chemokine receptor Ccr2 and proinflammatory cytokines Il1b and Tnfa in the DRG were significantly reduced to levels similar to those of sham-operated animals. This study provides the first evidence in an OA animal model that EGCG significantly slows OA disease progression and exerts a palliative effect. The online version of this article (doi:10.1186/s13075-014-0508-y) contains supplementary material, which is available to authorized users.