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[目的]通过白藜芦醇刺激体外培养的小鼠膝关节软骨细胞,探讨白藜芦醇通过PI3K/Akt信号通路对软骨细胞细胞外基质合成的影响。[方法]体外分离培养小鼠膝关节软骨细胞,采用HE染色、甲苯胺蓝染色和Ⅱ型胶原免疫荧光染色观察鉴定软骨细胞。根据作用药物的不同分为3组:空白对照组、白藜芦醇组(100μmol/L白藜芦醇)和LY294002组(100μmol/L白藜芦醇+25μmol/L LY294002)。采用RT-PCR检测软骨细胞中Akt1,Aggrecan、Collagen II的m RNA表达水平。[结果]荧光显微镜下可见软骨细胞胞核呈蓝色荧光,胞质呈红色荧光。显示Ⅱ型胶原染色阳性细胞数占90%以上,证实所培养细胞为软骨细胞。RT-PCR检测结果显示白藜芦醇组Akt1、Aggrecan、Collagen II m RNA表达量较空白组明显升高(P<0.05)。LY294002组Akt1、Aggrecan、Collagen II m RNA表达量较白藜芦醇组下降,但仍高于空白组(P<0.05)。[结论]白藜芦醇可以通过激活PI3K/Akt信号通路,增加软骨细胞细胞外基质合成,起到保护关节软骨的作用。
[Objective] To investigate the effect of resveratrol on the synthesis of extracellular matrix of chondrocytes through the PI3K / Akt signaling pathway by resveratrol stimulation of mouse knee chondrocytes. [Method] The mouse knee chondrocytes were isolated and cultured in vitro. The chondrocytes were identified by HE staining, toluidine blue staining and type Ⅱ collagen immunofluorescence staining. According to the different drugs, the rats were divided into 3 groups: blank control group, resveratrol group (100μmol / L resveratrol) and LY294002 group (100μmol / L resveratrol + 25μmol / L LY294002). The mRNA expression levels of Akt1, Aggrecan and Collagen II in chondrocytes were detected by RT-PCR. [Results] Under the fluorescence microscope, the nuclei of chondrocytes showed blue fluorescence and the cytoplasm showed red fluorescence. The results showed that more than 90% of type II collagen-positive cells were found, indicating that the cultured cells were chondrocytes. The results of RT-PCR showed that the expressions of Akt1, Aggrecan and Collagen II mRNA in resveratrol group were significantly higher than those in blank group (P <0.05). The expression of Akt1, Aggrecan and Collagen II mRNA in LY294002 group was lower than that in the resveratrol group, but still higher than that in the blank group (P <0.05). [Conclusion] Resveratrol can protect the articular cartilage by activating the PI3K / Akt signaling pathway and increasing the extracellular matrix synthesis of chondrocytes.