桃仁-红花药对对动静力失衡性大鼠椎间盘软骨终板细胞老化及凋亡的影响  

Effects of Taoren Honghua drug pair on aging and apoptosis of cartilage endplate in rats with cervical disc dynamic and static forces imbalance

作　　者：王艺儒 梁倩倩[2] 唐德志[2] 徐浩[2] 赵永见[2] 郑为超[1] WANG Yiru;LIANG Qianqian;TANG Dezhi(Medical Faculty,Anhui University of Science and Technology,Huainan 232001)

机构地区：[1]安徽理工大学医学院,淮南硕士研究生232001 [2]上海中医药大学脊柱病研究所

出　　处：《环球中医药》2018年第4期497-501,共5页Global Traditional Chinese Medicine

基　　金：国家自然科学基金(81273777)

摘　　要：目的观察桃仁-红花药对对动静力失衡性大鼠退变椎间盘软骨终板细胞老化及凋亡的影响。方法选取50只雄性Wistar大鼠,清洁级,随机分为五组,分别为对照组、假手术组、模型组、桃仁-红花药对组、美洛昔康组,每组10只。采用动静力失衡性大鼠椎间盘退变模型手术造模方法,对模型组、桃仁-红花药对组以及美洛昔康组进行手术造模,假手术组切开皮肤后不进行任何操作即缝合。造模3月后,桃仁-红花药对组、美洛昔康组分别给予桃仁-红花药对煎剂、美洛昔康悬浊液,每日一次灌胃;对照组、假手术组及模型组给予等量的生理盐水,每日一次灌胃,连续给药4周后一次性处死全部大鼠。取出大鼠完整C_(4/5)、C_(6/7)椎间盘,C_(4/5)椎间盘固定、切片,ABHO染色观察椎间盘组织形态学改变,免疫组化检测P21的表达,TUNEL荧光法检测软骨终板细胞凋亡;C_(6/7)椎间盘提取总mRNA,反转录后实时定量PCR检测P21 mRNA表达。结果模型组大鼠椎间盘组织形态学有明显退行性改变;对比于模型组,桃仁-红花药对组椎间盘软骨细胞凋亡数量显著减少(P<0.01);免疫组化及实时定量PCR检测显示,桃仁-红花药对组老化相关因子P21表达较模型组明显下降,差异均有统计学意义(P<0.01)。结论桃仁-红花药对可通过有效抑制老化相关基因P21表达,同时抑制椎间盘软骨终板细胞的凋亡,从而抑制椎间盘软骨的退变。Objective This study aimed at observing the effect of Taoren Honghua drug pair on apoptosis of degenerative intervertebral disc cartilage endplate in rats with static and dynamic imbalance.Methods Fifty male Wistar rats were randomly divided into control group,sham operation group,model group,Taoren Honghua drug pair group and Meloxicam group,10 rats in each group.Cervical disc dynamic and static forces imbalance of degeneration model was established in model group,Taoren Honghua drug pair group and Meloxicam group.Three months later,drug groups were taken by intragastric administration for four weeks,and other groups were given equivalent saline.The intact C4/5 and C6/7 intervertebral discs were removed,and the C4/5 intervertebral disc was fixed and sectioning.ABHO staining was used to observe the histomorphological changes of the intervertebral discs.The expression of P21 was detected by immunohistochemistry,and the apoptosis of cartilage endplate cells was detected by TUNEL fluorescence.The total mRNA was extracted from C6/7 intervertebral disc and the expression of P21 mRNA was detected by real-time quantitative PCR after reverse transcription.Results The histomorphology of the intervertebral disc in the model group has obvious degenerative changes.Compared with the model group,the number of chondrocyte apoptosis of intervertebral disc in Taoren Honghua drug pair group was decreased significantly(P<0.01);Immunohistochemistry and real-time quantitative PCR showed that the expression of aging related factor P21 in Taoren Honghua drug pair group was significantly lower than that in model group,and the difference was statistically significant(P<0.01).Conclusion Taoren Honghua drug pair can inhibit the expression of aging related gene P21,inhibit the apoptosis of intervertebral disc endplate cells,and inhibit the degeneration of intervertebral disc cartilage.

关 键 词：细胞凋亡 软骨终板 椎间盘退变 P21核糖核酸 细胞老化 

分 类 号：R285.5[医药卫生—中药学]