Effect and Interactions of Active Fractions in Yaotongning Capsule on Chondrocytes in Rat

ZHANG Li-guo; OUYANG Xiao-wen; WU Ting-ting; SHI Wan-zhong; NI Li-jun

doi:10.13422/j.cnki.syfjx.2014100151

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Effect and Interactions of Active Fractions in Yaotongning Capsule on Chondrocytes in Rat

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- Effect and Interactions of Active Fractions in Yaotongning Capsule on Chondrocytes in Rat
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 20, Issue 10, Pages: 151-155(2014)
- 作者机构：
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- DOI：10.13422/j.cnki.syfjx.2014100151
  CLC： R285.5
- Published：2014
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ZHANG Li-guo, OUYANG Xiao-wen, WU Ting-ting, et al. Effect and Interactions of Active Fractions in Yaotongning Capsule on Chondrocytes in Rat [J]. Chinese journal of experimental traditional medical formulae, 2014, 20(10): 151-155.
DOI：

ZHANG Li-guo, OUYANG Xiao-wen, WU Ting-ting, et al. Effect and Interactions of Active Fractions in Yaotongning Capsule on Chondrocytes in Rat [J]. Chinese journal of experimental traditional medical formulae, 2014, 20(10): 151-155. DOI： 10.13422/j.cnki.syfjx.2014100151.

摘要

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10 μg·L-1 IL-1β的DMEM培养基）及受试药物组（含5%FBS，0.02%吐温-80，10 μg·L-1 IL-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。

Abstract

Objective: To evaluate the in vitro effects and interactions of active fractions in Yaotongning capsule (YTNC) by utilizing interleukin-1β (IL-1β) activated chondrocytes. Method: Six samples were prepared by mixing different active fractions including alkaloids

flavonoids

saponins

volatile oil

hydrous extract and polysaccharides according to the formulating principle and the ratio of materials in YTNC. Chondrocytes were obtained from six seven-day rats using collagenase II

before they were divided into three groups:control group (Dulbecco's modification of Eagle's medium Dulbecco (DMEM) containing 5% fetal bovine serum (FBS) and 0.02% Tween-80)

model group (DMEM containing 5% FBS

0.02% Tween-80 and 10 μg·L-1 IL-1β) and sample group (DMEM containing 5% FBS

0.02% Tween-80

10 μg·L-1 IL-1β and 9.77-0.08 mg·L-1 samples). After 48 h

the proliferations of chondrocytes were examined through cell counting kit-8 (CCK8) assay

when the intracellular level of glycosaminoglycan (GAG) was detected by 1

9-dimethylmethylene blue staining using a glycosaminoglycans (GAG) assay kit. A quantitative evaluation method was exerted to investigate the effect of each sample on IL-1β activated chondrocytes. Result: The comparison of EC50 of YTNC sample and its partial prescriptions indicated that YTNC prescription had the most significant therapeutic effect on promoting proliferation and GAG synthesis in chondrocytes. Synergistic

antagonistic and additive action all could be observed followed by adding alkaloids

flavonoids

saponins

volatile oil

hydrous extract and polysaccharides to each partial prescription. Synergistic and additive effects between active fractions in YTNC prescription were observed

respectively. Conclusion: The therapeutic effect on osteoarthritis (OA) of YTNC without Chinese rice wine was obviously improved after the addition of Chinese rice wine

which indicated that the combination of active fractions in YTNC was reasonable.

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