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本文探讨了中药单体苦杏仁苷(amygdalin)联合羟基红花黄色素A(HSYA)对IL-1β诱导退变的大鼠椎间盘软骨终板细胞的影响,寻找可能的作用机制。从1月龄SD大鼠椎间盘中分离软骨终板并培养软骨终板细胞,经鉴定后选择第3代软骨终板细胞用于实验,随机分为正常组、诱导组、苦杏仁苷组、羟基红花黄色素A组和联合用药组。采用CCK-8法检测细胞增殖,流式检测细胞凋亡,Real-time PCR(RT-PCR)检测Aggrecan、Col 2α1、Col 10α1、MMP-13、IL-1βmRNA的表达,细胞免疫荧光检测II型胶原、X型胶原的表达。结果表明,与正常组比较,IL-1β诱导组软骨终板细胞增殖降低,细胞凋亡增加(P<0.05),下调Aggrecan、Col 2α1 mRNA的表达,上调Col 10α1、MMP-13、IL-1βmRNA的表达(P<0.05);同时II型胶原蛋白的表达降低、X型胶原蛋白的表达升高。与IL-1β诱导组比较,苦杏仁苷组、羟基红花黄色素A组和联合用药组均能在抑制软骨终板细胞凋亡的同时促进软骨终板细胞的增殖(P<0.05),上调Aggrecan、Col 2α1 mRNA的表达的同时下调Col 10α1、MMP-13、IL-1βmRNA的表达(P<0.05);促进II型胶原蛋白的表达增强、X型胶原蛋白的表达降低。联合用药组作用的效果明显优于苦杏仁苷组、羟基红花黄色素A组(P<0.05)。苦杏仁苷联合羟基红花黄色素A可抑制IL-1β诱导的大鼠椎间盘软骨终板软骨细胞发生退变,优于苦杏仁苷、羟基红花黄色素A单独用药。
This article explored the effect of amygdalin combined with hydroxysafflor yellow A (HSYA) on IL-1βinduced degenerative disc cartilage end-plate cells in rats, looking for a possible mechanism. The cartilage end-plates were isolated from the 1-month-old SD rat intervertebral disc and the cartilage end-plate cells were cultured. The third generation of cartilage end-plate cells were selected and used in the experiment. The cells were randomly divided into normal group, induction group, amygdalin group, Safflower A group and combination group. The cell proliferation was detected by CCK-8 assay and the apoptosis was detected by flow cytometry. The expression of Aggrecan, Col 2α1, Col 10α1, MMP-13 and IL-1βmRNA was detected by Real-time PCR Collagen, type X collagen expression. The results showed that compared with the normal group, IL-1βinduced the proliferation of cartilage end-plate cells decreased and the apoptosis increased (P <0.05), down-regulated the expression of Aggrecan and Col 2α1 mRNA and up-regulated the expression of Col 10α1, MMP-13 and IL- (P <0.05). At the same time, the expression of type II collagen decreased and the expression of type X collagen increased. Compared with IL-1βinduced group, both amygdalin group, hydroxysafflor yellow A group and combined treatment group could promote the proliferation of cartilage end-plate cells (P <0.05) and up-regulate the apoptosis of cartilage end-plate cells Aggrecan and Col 2α1 mRNA, and down-regulated the expression of Col 10α1, MMP-13 and IL-1βmRNA (P <0.05), promoted the expression of type II collagen and decreased the expression of type X collagen. The effect of combined treatment group was significantly better than that of amygdalin and hydroxysafflor yellow A (P <0.05). Amygdalin combined with hydroxysafflor yellow A can inhibit IL-1β-induced rat disc degeneration of cartilage end-plate chondrocytes, superior to amygdalin, hydroxysafflor yellow A alone.