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目的:评价腰痛宁胶囊中各活性部位对白介素1β(IL-1β)诱导大鼠软骨细胞的影响及其相互作用。方法:根据腰痛宁组方原则,将腰痛宁组方药材中的黄酮、皂苷、挥发油(含水提液)、多糖等活性部位依次加入马钱子与麻黄生物碱中,得到6个腰痛宁拆方及全方样品。取7日龄SD大鼠6只,采用Ⅱ型胶原酶消化法分离大鼠软骨细胞,接种24 h后随机分空白对照组(含5%小牛血清(FBS)及0.02%吐温-80的DMEM培养基)、模型组(含5%FBS,0.02%吐温-80及10μg·L-1IL-1β的DMEM培养基)及受试药物组(含5%FBS,0.02%吐温-80,10μg·L-1IL-1β及9.77~0.08 mg·L-1受试药物的DMEM培养基),给药48 h后用CCK8及亚甲基蓝染色法测定各样品对大鼠软骨细胞增殖及葡萄糖胺聚糖蛋白(GAG)合成的影响。同时采用最小二乘优化方法,计算各样品的半数有效浓度(EC50)叠加值,将其与EC50实验值进行比较,以分析各活性部位间的相互作用关系。结果:较腰痛宁拆方样品,腰痛宁组方及全方在考察浓度范围内均对IL-1β诱导的软骨细胞具有较显著的促增殖及促进GAG合成的作用。各拆方样品中可观察到不同程度的协同、拮抗和叠加作用,其中腰痛宁全方样品中各活性部位间的协同及叠加作用更为显著。结论:药引黄酒可显著提高腰痛宁药材组方对大鼠骨关节炎的治疗作用,显示了腰痛宁处方的合理性及其促进软骨细胞增殖及GAG合成的优势。
OBJECTIVE: To evaluate the effects of various active sites in Yaotongning Capsules on rat chondrocytes induced by interleukin-1β (IL-1β) and their interaction. Methods: According to the principles of Yaotongning prescription, the active components of flavonoids, saponins, volatile oil (aqueous extract) and polysaccharides in Yaotongning prescription herbs were added into the alkaloids of Strychnos nux vomica and Ephedra, And all samples. Chondrocytes were isolated from 6-day-old SD rats by type Ⅱ collagenase digestion and randomly divided into blank control group (containing 5% fetal bovine serum (FBS) and 0.02% Tween-80 DMEM medium), model group (DMEM medium containing 5% FBS, 0.02% Tween-80 and 10μg · L-1IL-1β) and test drug group containing 5% FBS, 0.02% Tween-80, 10μg · L-1IL-1β and 9.77 ~ 0.08 mg · L-1 of the test drug in DMEM medium), 48 h after administration, CCK8 and methylene blue staining were used to determine the effect of each sample on rat chondrocyte proliferation and glycosaminoglycan Protein (GAG) synthesis. At the same time, the least square method was used to calculate the half value effective concentration (EC50) of each sample, and compared with the EC50 values to analyze the interaction between the active sites. Results: Compared with those of Yaotseining decoction, the group of Yaotongning Decoction and all prescriptions all had significant effects on IL-1β-induced chondrocytes proliferation and GAG synthesis in the concentration range. Various demixing samples were observed in different degrees of synergy, antagonism and superposition effect, in which Liaotongning all samples in the active site synergy and superimposition effect is more significant. Conclusion: Yin-Yin Huangjiu can significantly improve the therapeutic effect of Yaotongning herbs on osteoarthritis in rats, which shows the rationality of Yaotongning Prescription and its advantage of promoting chondrocyte proliferation and GAG synthesis.