BACKGROUND:Studies have shown that Clematis chinensis Osbeck can promote articular chondrocyte proliferation, maintain and promote the synthesis of cartilage proteoglycan and type II col agen in chondrocytes. Low-intensity pulsed ultrasound can effectively improve the carticular chondrocyte proliferation, increase the cellmembrane permeability, and promote chemicals or genes delivery, so as to improve the biological effects of chemicals. OBJECTIVE:To observe the effect of low-intensity pulsed ultrasound mediated Clematis chinensis Osbeck on the proliferation of cartilage cells and the expression of type II col agen and transforming growth factor (TGF)-β1 of rabbit knee articular chondrocytes cultured in vitro, and explore the effect and mechanism of Clematis chinensis Osbeck mediated by low-intensity pulsed ultrasound on cartilage damage. METHODThe rabbit knee articular chondrocytes were isolated and cultured in vitro. cells of the second generation in logarithmic growth phase were randomly divided into four groupcontrol group, low-intensity pulsed ultrasound (LIPUS) group, Clematis chinensis Osbeck (CCO) group, and CCO+LIPUS group. After cells were cultured under different conditions for 3 days, cellproliferation was measured with cellCount Kit-8 assay. The secretion of type II col agen was detected by cytochemical staining method, and the expression of TGF-β1 was assayed by western blot analysis. RESULTS AND CONCLUSION:The number of cells in the other three groups were significantly higher than that in control group at 3 days after culture (P<0.05), while the CCO+LIPUS group had the most cells. Cytochemical staining showed that the area and absorbance value of type II col agen in cartilage cells were significantly increased in CCO+LIPUS group, compared with control group (P<0.05), and the difference with control group was larger than the amount of that of LIPUS group and CCO group with control group. Western blot analysis showed that TGF-β1 was significantly expressed in CCO+LIPUS group, and the relative gray value was significantly higher than the other groups (P<0.05). Both low-intensity pulsed ultrasound and Clematis chinensis Osbeck can promote the proliferation of cartilage cells and expression of type II col agen and t-β1 of rabbit knee articular chondrocytes cultured in vitro. The combined method could produce synergistic effects.%背景：研究显示威灵仙可促进软骨细胞增殖，维持和促进软骨细胞合成蛋白多糖和Ⅱ型胶原；低强度脉冲超声能有效促进软骨细胞增殖、提高细胞膜的通透性，促进药物或基因的传递，从而提高药物的生物学效应。 目的：观察低强度脉冲超声介导威灵仙对体外培养的兔膝关节软骨细胞增殖、Ⅱ型胶原及转化生长因子β1表达的影响，探讨低强度脉冲超声介导威灵仙在软骨损伤修复中的作用及机制。 方法：体外分离培养兔膝关节软骨细胞，取处于对数生长期的2代细胞，随机分为4组：对照组，低强度脉冲超声组，威灵仙组，威灵仙+低强度脉冲超声组，干预3 d后分别采用CCK-8比色法检测软骨细胞增殖情况，细胞化学染色法分析Ⅱ型胶原分泌情况及Western blot检测转化生长因子β1的表达情况。 结果与结论：干预培养3 d后，与对照组比较，其他3组细胞数量均显著增加(P <0.05)，威灵仙+低强度脉冲超声组细胞数量最多；威灵仙+低强度脉冲超声组软骨细胞Ⅱ型胶原表达面积、吸光度值均显著大于对照组(P <0.05)，且其差值大于低强度脉冲超声组、威灵仙组与对照组差值之和；威灵仙+低强度脉冲超声组转化生长因子β1表达水平显著，其相对灰度值显著高于其他各组(P <0.05)。结果提示低强度脉冲超声、威灵仙均可促进体外培养兔膝关节软骨细胞增殖及Ⅱ型胶原、转化生长因子β1的分泌，二者联用具有一定的协同作用。
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