补肾活血方含药血清对离体兔脊柱运动节段压力退变模型的影响  

Effect of serum of Bushen Huoxue prescription(补肾活血方) on rabbit with intervertebral disc motion segments in vitro culture

作　　者：展嘉文[1] 朱立国[2] 冯敏山[2] 王尚全[1] 张平[3] ZHAN Jia-wen;ZHU Li-guo;FENG Min-shan;WANG Shang-quan;ZHANG Ping(The Second Department of Spinal,Wangjing Hospital of China A cademy of Chinese Medical Sciences,Beijing 100102,China)

机构地区：[1]中国中医科学院望京医院骨伤综合科,北京100102 [2]中国中医科学院望京医院脊柱二科,北京100102 [3]中国中医科学院望京医院病理科,北京100102

出　　处：《中国骨伤》2018年第7期627-634,共8页China Journal of Orthopaedics and Traumatology

基　　金：国家自然科学基金项目(编号:81774330);国家中医药管理局国家中医临床研究基地业务建设科研专项课题(编号:JDZX2015274);国家体育总局中医特色技术在体育运动中的应用(编号:HXKT2017001)~~

摘　　要：目的:观察补肾活血方含药血清对离体兔脊柱运动节段压力退变模型的影响。方法 :将24只4～6月龄体重为2.5～3 kg的新西兰白兔,根据随机数字表分为含药组与对照组,每组各7只,处死后在无菌条件下分别取出脊柱运动节段各28个,均放入脊柱运动节段离体加载和培养装置中进行培养,其中含药组与对照组培养液中分别含有10%补肾活血方含药血清与10%无药血清,于培养前及培养后第3、7、14天,两组各取7个椎间盘分别进行组织形态学、蛋白多糖PAS/AB特染、Ⅱ型胶原免疫组化和Agg与Col2aI的Real-time PCR检测。结果:组织形态学检测示培养1周内含药组较对照组形态维持更好;PAS/AB检测示两组蛋白多糖含量均减少;免疫组化示含药组Ⅱ型胶原含量3 d时较对照组明显升高(P<0.05);1周后虽明显降低,但仍较对照组显著(P<0.05);RT-PCR显示两组培养3 d后Agg表达较术前显著下降(P<0.05)(P<0.01),两组7 d与14 d时比较差异无统计学意义(P>0.05);两组3 d时Col2aI表达均明显上调,且含药组表达更加显著(P<0.01),7 d后两组虽明显下降(P<0.05),但含药组比对照组仍表达显著(P<0.05),14 d时两组表达水平均较低,差异无统计学意义(P>0.05)。结论 :补肾活血方含药血清短期内能一定程度上延缓椎间盘退变,可能与该方改善循环、抑制炎性、调节细胞外基质有关,提示本方对椎间盘退变有早期防治作用。Objective：To compare effect of serum of Bushen Huoxue prescription（补肾活血方） on rabbit with intervertebral disc motion segments in vitro culture Methods：Twenty four New Zealand white rabbits aged from 4 to 6 months and weighted from 2.5 to 3 kg were divided into medicated group and control group,7 in each group. Rabbits were excuted under condition of asepsis,28 spinal motion segments were taken out in each group,and segments were loaded in spinal motion segments in vitro and cultured in culture apparatus. Nutrient solution of medicated group contain 10% serum of Bushen Huoxue prescription（补肾活血方）,and 10% blank serum in control group. Seven discs between two groups were taken out and observed by histomorphology,proteoglycan PAS/AB stining,collagenⅡimmunohistochemical staining,AGG,Col2aI by PCR test before culture and on the 3 th,7 th and 14 th day after culture. Results：Histomorphology results showed the formation of medicated group was better than that of control group at 1 week after culture. PAB/AB test results showed content of poteoglycan between two groups were decreased. Imunohistochemical results showed content of collagen Ⅱ in medicated group were obviously increased than that of control group at 3 days,but decreased obviously at 1 week than that of control group. PCR results showed expression of Agg in medicated group was obviously decreased than that of control group,but no statistical significance between two groups on the 7 th and 14 th day. Expression of Col2aI between two groups at 3 days were increased,and medicated group increased obviously more,while there were no significant difference between two groups. Conclusion：Serum of Bushen Huoxue prescription（补肾活血方） could delay intervertebral disc degeneration at short time for it is relate with improving circulation,inhibiting inflammatory,regulating extracellular matrix,so the prescription plays an important role in early prevention and treatment for intervertebral disc degeneration.

关 键 词：兔 脊柱 椎间盘退行性变 血清 

分 类 号：R285.5[医药卫生—中药学] R-332[医药卫生—中医学]