河蚌葡聚糖对半月板组织损伤及病理性过度肥大和钙化的影响

发布时间：2018-06-26 11:16

本文选题：河蚌葡聚糖 + 异常压力　；参考：《南方医科大学学报》2017年04期

【摘要】：目的观察中药单体河蚌葡聚糖(HBP-A)对因异常力学刺激引起的半月板组织损伤及病理性过度肥大和钙化相关基因蛋白表达的影响。方法将牛的半月板组织切割成统一大小的圆柱形,施加使组织发生可压缩25%的异常压力刺激3 h,将半月板随机分为对照组、力学压力刺激3 h组、药物组(力学压力刺激3 h+0.6 mg/mL HBP-A培养72 h)。采用细胞活性检测方法观察各组半月板细胞的成活率,二甲基亚甲基蓝检测培养基中蛋白多糖的含量以此来定量各组半月板组织蛋白多糖的丢失程度。通过RT-qPCR和Westernblot来检测HBP-A对病理性过度肥大和钙化相关因子ANKH、ENPP1、ALP和MMP13、IL-1在mRNA和蛋白水平上表达的影响。结果在药物组,细胞死亡(红色荧光)数量虽然多于正常对照组,但仍明显低于力学压力刺激3 h组。在培养液中,力学压力刺激3 h组蛋白多糖的含量是正常对照组的1.4倍(P0.05),是药物组蛋白多糖含量的1.3倍(P0.05),说明该药物明显抑制了半月板蛋白多糖的丢失。同时,在药物组,半月板病理性过度肥大和钙化相关因子ANKH、ENPP1、ALP和MMP13、IL-1在mRNA和蛋白水平上的表达也明显低于力学压力刺激3 h组(P0.05)。结论 HBP-A能够明显地抑制因异常力学刺激引起的半月板组织损伤(细胞死亡、多糖丢失)及病理性过度肥大和钙化相关基因蛋白的表达,可能是治疗膝骨关节炎的潜在性药物。
[Abstract]:Objective to observe the effect of Chinese medicine monomeric mussel dextran (HBP-A) on meniscus tissue injury and pathological hypertrophy and calcification-related gene protein expression induced by abnormal mechanical stimulation. Methods the meniscus tissue of cattle was cut into a cylindrical shape of uniform size. The meniscus was subjected to a compressible 25% abnormal pressure stimulation for 3 h. The meniscus was randomly divided into control group and mechanical pressure stimulation group for 3 h. Drug group (mechanical pressure stimulation 3 h 0.6 mg / mL HBP-A culture 72 h). The survival rate of meniscal cells and the content of proteoglycan in culture medium were measured by using the method of cell activity test to quantify the degree of loss of proteoglycan in meniscus tissue of each group. The effects of HBP-A on the expression of ALP and MMP13 IL-1 in the pathological hypertrophy and calcification related factors ANKHH, ENPP1, ALP and MMP13 were detected by RT-qPCR and Western blot. Results the number of cell death (red fluorescence) in the drug group was higher than that in the normal control group, but still significantly lower than that in the mechanical pressure stimulation group for 3 h. In the culture medium, the content of histone proteoglycan stimulated by mechanical pressure for 3 h was 1.4 times of that of the normal control group (P0.05) and 1.3 times of the drug histone polysaccharide content (P0.05), indicating that the drug significantly inhibited the loss of the meniscus proteoglycan. At the same time, the expression of ALP and MMP13 IL-1 in the pathological hypertrophy and calcification of meniscus in the drug group was significantly lower than that in the group stimulated by mechanical pressure for 3 h (P0.05). Conclusion HBP-A can significantly inhibit the meniscus injury (cell death, polysaccharide loss) and the expression of pathological hypertrophy and calcification related gene protein induced by abnormal mechanical stimulation. HBP-A may be a potential drug for the treatment of knee osteoarthritis.
【作者单位】：上海中医药大学附属曙光医院石氏伤科医学中心;上海市中医药研究院骨伤科研究所;上海中医药大学附属曙光医院肿瘤科;中国中医科学院望京医院;
【基金】：国家自然科学基金(81473702,81674005);国家自然科学基金青年科学基金(81503596,81202707,81001528) 北京市自然科学基金青年项目(7164313) 中医骨伤科学”国家重点学科(100508) 上海市中医药领军人才建设项目(2012-63-15) 海派中医流派传承研究基地(ZY3-CCCX-1-1003) 上海市教育委员会高峰高原建设中医骨伤团队张江高科技园区中医药发展扶持项目(pzy2015-1)~~
【分类号】：R684

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