补肾壮筋汤联合盐酸氨基葡萄糖对大鼠膝软骨细胞增殖及凋亡影响的研究

目的研究中药经方补肾壮筋汤联合西药盐酸氨基葡萄糖,制备相应浓度的含药血清,作用于大鼠膝软骨细胞,为膝骨性关节炎的治疗提供新的理论依据及方法。方法将SPF级大鼠(32只)随机分为4组,分别以生理盐水、补肾壮筋汤、盐酸氨基葡萄糖、补肾壮筋汤联合盐酸氨基葡萄糖,对大鼠进行药物灌胃1周,制备相应浓度的含药血清作用于离体培养的大鼠膝软骨细胞,通过MTT法检测不同浓度(0. 6%、1. 2%、2. 4%)含药血清对软骨细胞增殖活力、流式细胞分析仪检测细胞凋亡率、Western blot检测软骨细胞中II型胶原蛋白的表达情况。结果 MTT法检测示,对大鼠膝软骨细胞的促进增殖作用中,中西医结合组优于其他各组,差异具有统计学意义(P0. 01);流式细胞仪检测细胞凋亡率示,中西结合血清组能够明显降低软骨细胞凋亡率,与其他各组比较差异具有统计学意义(P0. 01); Western blot示,中西医结合血清组促进软骨细胞中II型胶原蛋白表达明显高于其他各组,差异具有统计学意义(P0. 01)。结论中西医药物联合应用对大鼠膝软骨细胞增殖作用明显高于中药组及西药组,并能够促进软骨细胞中II型胶原蛋白的表达,降低大鼠膝软骨细胞凋亡的发生,从而起到骨性关节炎的治疗作用。

Effect of the reinforcing kidney and strengthening tendon decoction combined with glucosamine hydrochloride on the proliferation and apoptosis of knee chondrocytes in rats

Objective To study the effect of the corresponding concentration of drug-containing serum of Chinese medicine reinforcing kidney and strengthening tendon decoction combined with Western medicine glucosamine hydrochloride on rat knee chondrocytes, and to provide a new theoretical basis and method for the treatment of knee osteoarthritis. Methods Thirty-two SPF grade rats were randomly divided into 4 groups. Rats received normal saline, reinforcing kidney and strengthening tendon decoction, glucosamine hydrochloride, reinforcing kidney and strengthening tendon decoction combined with glucosamine hydrochloride, respectively. The rats were intragastrically administrated for 1 week. The drug-containing serum was applied to rat knee chondrocytes cultured in vitro. The proliferation of chondrocytes at different concentrations (0.6%, 1.2%, 2.4%) of drug-containing serum was detected with MTT assay. The apoptosis rate was detected with a flow cytometry analyzer. The expression of type II collagen in chondrocytes was detected with Western blotting. Results MTT assay results showed that the proliferation of rat knee chondrocytes in Chinese and Western drug combination group was better than that in other groups (P<0.01). The difference was statistically significant. Flow cytometry results showed that apoptosis rate of chondrocytes in Chinese and Western combination drug-conditioned serum group was significantly reduced, compared to that in other groups, and the difference was statistically significant (P<0.01). Western blotting results showed that in Chinese and Western medicine combination serum group, the expression of type II collagen in chondrocytes was significantly higher than that in other groups (P<0.01), and the difference was statistically significant. Conclusion The rat knee chondrocyte proliferation is significantly higher in the combination of Chinese and Western medicine group than in Chinese medicine group or Western medicine group. The combination of Chinese and Western medicine promotes the expression of type II collagen in chondrocytes, reduces the occurrence of apoptosis in rat knee chondrocyte, and thus plays a role in the treatment of osteoarthritis.