益气化瘀补肾方对退变椎间盘细胞蛋白聚糖和Ⅹ型胶原mRNA表达的影响  

Effects of Chinese herbal medicine Yiqi Huayu Bushen Recipe on expressions of aggrecan and type Ⅹ collagen mRNAs in cells from degenerated human intervertebral discs

作　　者：李具宝[1,2] 赵红云[3] 赵永见[2] 周泉[2] 李宁[1,2] 许鹏[4] 施杞[2] 王拥军[2] 

机构地区：[1]上海中医药大学龙华医院临床医学院,上海200032 [2]上海中医药大学脊柱病研究所,上海200032 [3]上海中医药大学龙华医院骨伤科,上海200032 [4]第二军医大学长征医院骨科,上海200003

出　　处：《中西医结合学报》2011年第10期1138-1144,共7页Journal of Chinese Integrative Medicine

基　　金：上海市高校创新团队计划资助项目(No.沪教委科[2009]6号);国家重点基础研究发展计划(973计划)资助项目(No.2010CB530400);国家教育部高级人才项目长江学者特聘教授计划项目(No.教人[2009]17号);国家教育部高等学校博士点基金资助项目(No.20093107110003);国家自然科学基金资助项目(No.30772811;No.81072831)

摘　　要：目的:观察益气化瘀补肾方血清对脊髓型颈椎病患者退变椎间盘细胞蛋白聚糖和Ⅹ型胶原mRNA表达的影响。方法:取6例确诊为脊髓型颈椎病的住院患者行颈椎减压融合手术摘除的退变椎间盘组织块,采用组织块法培养原代细胞;SD大鼠10只,灌胃制备大鼠益气化瘀补肾方含药血清。取传代第1代的退变椎间盘细胞,分为益气化瘀补肾方含药血清高浓度组(20%)、中浓度组(10%)、低浓度组(5%)及相应体积比浓度氯化钠溶液血清对照组。MTT法检测不同浓度的益气化瘀补肾方血清对椎间盘细胞增殖的影响,实时荧光定量聚合酶链反应检测不同给药时间点各组椎间盘细胞蛋白聚糖和Ⅹ型胶原mRNA的表达。结果:第1代退变椎间盘细胞传代第6天时细胞增殖达到顶峰,8d后开始下降,故选择细胞传代第6天为给药干预时间点。与对照血清比较,不同浓度的益气化瘀补肾方血清可促进退变椎间盘细胞的增殖(P<0.01),提高退变椎间盘细胞蛋白聚糖mRNA的表达(P<0.01),并下调Ⅹ型胶原mRNA的表达(P<0.01),且以中浓度的作用最为明显(P<0.05)。结论:益气化瘀补肾方通过调节退变椎间盘的胶原及蛋白多糖的表达而产生有效防治脊髓型颈椎病椎间盘退变的作用。Objective：To investigate the effects of serum containing Yiqi Huayu Bushen Recipe,a compound Chinese herbal medicine,on expressions of aggrecan and type Ⅹ collagen mRNAs of the degenerated intervertebral disc cells in patients with cervical spondylotic myelopathy.Methods：Six inpatients diagnosed as cervical spondylotic myelopathy and undergoing cervical decompression and fusion surgery to remove the degenerated disc tissue were enrolled in this study.Primary intervertebral disc cells were isolated using small tissue mass method.Serum containing Yiqi Huayu Bushen recipe were obtained after SD rats were fed with Yiqi Huayu Bushen recipe for 3 days.MTT method was used to detect proliferation of the first-passage cells isolated from degenerated intervertebral discs and growth curve was drawn.In the following tests,the first-passage cells were cultured with the sera containing drugs and sodium chloride（NaCl）,respectively.Proliferation of intervertebral disc cells was detected by MTT method and expressions of aggrecan and type Ⅹ collagen mRNAs were detected by real-time polymerase chain reaction.Results：The growth curve of the first-passage cells of the degenerated intervertebral discs showed that the time of cell proliferation peak was the sixth day.Compared with the same concentrations of NaCl solution,serum containing Yiqi Huayu Bushen Recipe at 5%,10% and 20% promoted cell proliferation and expression of aggrecan mRNA and decreased expression of type Ⅹ collagen mRNA（P0.01）.10% serum containing Yiqi Huayu Bushen recipe was more effective than 5% and 20%.Conclusion：Yiqi Huayu Bushen Recipe inhibits degeneration of the intervertebral disc cells by regulating collagen and proteoglycan expressions.

关 键 词：颈椎病 中草药 椎间盘 蛋白聚糖 胶原Ⅹ型 

分 类 号：R285[医药卫生—中药学]