补肾益气行血法对人软骨细胞基质金属蛋白酶13表达的影响

The Regulating effects of Reinforcing kidney and nourishing qi and blood on gene expression of metalloproteinase-13 in human hyaline chondrocytes

目的:观察补肾益气行血中药复方含药血清对MMP-13、Ⅱ型胶原表达的调控作用及相关意义。方法:将体外培养的人软骨细胞随机分为5组:对照组(正常血清)、低剂量IL-1组(10μg/ml IL-1β)、高剂量IL-1组(100μg/ml IL-1β)、低剂量IL-1加中药组(10μg/ml IL-1β加含药血清)、高剂量IL-1加中药组(100μg/ml IL-1β加含药血清),应用逆转录-聚合酶链反应(RT-PCR)及实时荧光定量PCR,比较补肾益气行血中药复方含药血清在不同剂量IL-1β作用下对MMP-13、Ⅱ型胶原表达的调控作用。结果:中药含药血清组均有抑制MMP-13的表达作用,对照组表达阳性强度高于中药组,中药组Ⅱ型胶原表达阳性强度高于对照组,统计学均有明显差异。不同剂量IL-1组均能促进MMP-13的表达,对照组阳性强度低于IL-1组,IL-1组Ⅱ型胶原表达低于对照组,统计学均有明显差异。结论:体外条件下,补肾益气行血中药含药血清能够抑制炎性因子IL-1诱导的MMP-13表达;能对抗MMP-13抑制软骨细胞Ⅱ型胶原合成,具有保护软骨细胞的作用。

Objective：To investigate the role of Reinforcing kidney and nourishing qi and blood in regulating the gene expression of ma- trix metalloproteinase-13 （MMP-13） in the human hyaline chondrocytes. Methods ：The human hyaline chondrocytes harvested enzymati- cally and cultured in DMEM supplemented with 20% fetus calf serum were divided into 7 groups：Group 1 was used as a contol（ pure blood serum）, Low IL-1 [3group （101xg/ml IL-113）, High IL-113group （100μg/ml IL-113）, Low IL-113 and Chinese medicine-conditioned serum group（ 10μg/ ml IL-113and Chinese medicine-conditioned serum）, High IL-1 and Chinese medicine-conditioned serum group （100μg/ ml IL-113and Chinese medicine-conditioned serum）. The MMP-13 and Collagen type Ⅱ levels of passaged human hyaline chondrocytes from above five groups were assessed by reverse transcription-polymerase chain reaction （RT-PCR） and real-time fluores- cent quantitative PCR. Result ：The group of Chinese medicine could restrain the gene expression of MMP-13, control group has higher expression than Chinese medicine group. The Chinese medicine group has higher collagen type 11 expression. It is difference in statis- tics. Low and high IL-113 group both promote the gene expression of MMP-13, control group has lower expression than Chinese medicine group. The IL-1β group has lower collagen type Ⅱ expression. It is difference in statistics. Conclusion. In vitro, Reinforcing kidney and nourishing qi and blood -conditioned serum can inhibit the gene expression of MMP-13 induced by IL-1β, increase collagen type Ⅱ , and potect human hyaline chondrocytes.