六味地黄丸对兔椎间盘退变模型椎间盘组织中Ⅰ、Ⅱ型胶原表达的影响

发布时间：2017-12-31 21:04

  本文关键词：六味地黄丸对兔椎间盘退变模型椎间盘组织中Ⅰ、Ⅱ型胶原表达的影响　出处：《中医正骨》2016年08期 　论文类型：期刊论文

  更多相关文章： 椎间盘退行性变 六味地黄丸 胶原Ⅰ型 胶原Ⅱ型 兔 动物实验

【摘要】：目的:观察六味地黄丸对兔椎间盘退变模型椎间盘组织中Ⅰ、Ⅱ型胶原表达的影响。方法:将80只新西兰兔随机分为空白组、假手术组、模型组和六味地黄组,每组20只。模型组、六味地黄组手术暴露L4~5和L5~6椎间隙,随机选择1个椎间盘注射肿瘤坏死因子α进行椎间盘退变造模,并以咬骨钳咬除该椎间盘上位椎体横突进行标记;假手术组经相同手术入路暴露L4~5和L5~6椎间隙,不做任何处理后缝合;空白组不进行任何手术干预。造模术后3 d开始,六味地黄组按26 mg·kg-1以六味地黄胶囊混悬液灌胃,其余3组以等量生理盐水灌胃,每天1次,共8周。分别于药物干预开始后2、4、6、8周,从各组随机选取5只兔子处死,模型组和六味地黄组手术取出造模椎间盘,空白组和假手术组随机选择L4~5或L5~6椎间盘取出。分别采用聚合酶链式反应法和Western Blot法测定椎间盘组织中Ⅰ、Ⅱ型胶原的mRNA和蛋白表达水平。结果:药物干预2、4、6、8周后,4组兔子椎间盘组织Ⅰ型胶原mRNA表达水平比较,组间差异均有统计学意义[(0.064 8±0.009 8),(0.068 6±0.012 7),(0.192 0±0.040 6),(0.124 6±0.012 0),F=49.752,P=0.000;(0.066 0±0.010 0),(0.077 1±0.011 8),(0.252 4±0.039 9),(0.164 1±0.018 1),F=79.537,P=0.000;(0.071 2±0.011 2),(0.089 1±0.008 1),(0.326 3±0.028 5),(0.176 3±0.015 0),F=236.726,P=0.000;(0.094 1±0.012 0),(0.155 0±0.003 8),(0.451 4±0.039 6),(0.205 8±0.018 1),F=201.055,P=0.000];模型组Ⅰ型胶原mRNA表达水平高于空白组、假手术组和六味地黄组(P=0.000,P=0.015,P=0.002,P=0.000;P=0.013,P=0.002,P=0.000,P=0.015;P=0.002,P=0.012,P=0.000,P=0.000)。空白组和假手术组的Ⅰ型胶原mRNA表达水平各时间点间比较,差异均无统计学意义(F=50.563,P=0.132;F=80.352,P=0.634);模型组和六味地黄组的Ⅰ型胶原mRNA表达水平各时间点间比较,差异均有统计学意义(F=193.635,P=0.000;F=284.736,P=0.000),Ⅰ型胶原mRNA表达水平均逐渐增高(P=0.004,P=0.002,P=0.000;P=0.000,P=0.003,P=0.001)。药物干预2、4、6、8周后,4组兔子椎间盘组织Ⅱ型胶原mRNA表达水平比较,组间差异均有统计学意义[(0.042 7±0.008 0),(0.041 2±0.005 8),(0.011 6±0.002 1),(0.031 7±0.005 6),F=42.696,P=0.000;(0.038 8±0.004 3),(0.037 3±0.004 3),(0.011 5±0.001 8),(0.031 1±0.003 5),F=108.110,P=0.000;(0.030 3±0.005 7),(0.025 9±0.008 3),(0.007 9±0.002 7),(0.017 2±0.002 1),F=52.436,P=0.000;(0.029 3±0.006 9),(0.023 7±0.004 6),(0.005 3±0.001 0),(0.014 8±0.001 8),F=51.375,P=0.000];模型组Ⅱ型胶原mRNA表达水平低于空白组、假手术组和六味地黄组(P=0.002,P=0.001,P=0.000,P=0.001;P=0.001,P=0.000,P=0.000,P=0.001;P=0.001,P=0.002,P=0.013,P=0.000)。空白组和假手术组的Ⅱ型胶原mRNA表达水平各时间点间比较,差异均无统计学意义(F=70.463,P=0.122;F=90.362,P=0.089);模型组和六味地黄组的Ⅱ型胶原mRNA表达水平各时间点间比较,差异均有统计学意义(F=110.364,P=0.001;F=86.362,P=0.004),模型组和六味地黄组的Ⅱ型胶原mRNA表达水平均逐渐降低(P=0.002,P=0.005,P=0.003;P=0.000,P=0.001,P=0.000)。药物干预2、4、6、8周后,4组兔子椎间盘组织Ⅰ型胶原蛋白表达水平比较,组间差异均有统计学意义[(0.575 9±0.135 6),(0.599 5±0.037 4),(0.620 0±0.072 0),(0.609 6±0.032 1),F=9.288,P=0.001;(0.593 9±0.108 0),(0.647 1±0.044 8),(0.807 0±0.061 6),(0.659 9±0.105 5),F=28.883,P=0.000;(0.614 0±0.224 6),(0.685 6±0.066 5),(1.129 1±0.097 4),(0.700 1±0.086 5),F=34.957,P=0.000;(0.614 8±0.139 3),(0.742 3±0.165 3),(1.322 4±0.351 7),(0.806 0±0.094 3),F=3.296,P=0.048];模型组Ⅰ型胶原蛋白表达水平高于空白组、假手术组和六味地黄组(P=0.000,P=0.015,P=0.002,P=0.000;P=0.013,P=0.002,P=0.000,P=0.015;P=0.002,P=0.012,P=0.000,P=0.000)。空白组和假手术组的Ⅰ型胶原蛋白表达水平各时间点间比较,差异均无统计学意义(F=37.485,P=0.365;F=10.376,P=0.583);模型组和六味地黄组的Ⅰ型胶原蛋白表达水平各时间点间比较,差异均有统计学意义(F=70.362,P=0.000;F=6.375,P=0.001),模型组和六味地黄组的Ⅰ型胶原蛋白表达水平均逐渐增高(P=0.005,P=0.002,P=0.000;P=0.004,P=0.001,P=0.000)。药物干预2、4、6、8周后,4组兔子椎间盘组织Ⅱ型胶原蛋白表达水平比较,组间差异均有统计学意义[(0.605 8±0.066 5),(0.550 0±0.117 9),(0.278 0±0.053 2),(0.498 6±0.149 1),F=10.224,P=0.001;(0.553 7±0.126 5),(0.523 4±0.078 4),(0.258 2±0.037 8),(0.479 7±0.090 8),F=11.627,P=0.000;(0.546 8±0.120 8),(0.494 8±0.139 8),(0.219 0±0.099 2),(0.440 5±0.052 7),F=13.543,P=0.003;(0.494 8±0.042 5),(0.433 7±0.061 9),(0.131 9±0.012 8),(0.392 9±0.107 0),F=13.979,P=0.000];模型组Ⅱ型胶原蛋白表达水平低于空白组、假手术组和六味地黄组(P=0.002,P=0.001,P=0.000,P=0.001;P=0.001,P=0.000,P=0.000,P=0.001;P=0.001,P=0.002,P=0.013,P=0.000)。空白组和假手术组的Ⅱ型胶原蛋白表达水平各时间点间比较,差异均无统计学意义(F=17.364,P=0.092;F=15.573,P=0.175);模型组和六味地黄组的Ⅱ型胶原蛋白表达水平各时间点间比较,差异均有统计学意义(F=17.753,P=0.001;F=13.674,P=0.000),模型组和六味地黄组的Ⅱ型胶原蛋白表达水平均逐渐降低(P=0.002,P=0.004,P=0.001;P=0.000,P=0.001,P=0.000)。结论:六味地黄丸能适度下调兔椎间盘退变模型椎间盘组织中Ⅰ型胶原的表达、上调Ⅱ型胶原的表达,可在一定程度上延缓椎间盘退变进程。
[Abstract]:Objective: To observe the effect of Six Ingredient Rehmannia Pill on rabbit model of intervertebral disc degeneration of intervertebral disc tissue, expression of type II collagen. Methods: 80 New Zealand rabbits were randomly divided into control group, sham operation group, model group and six Dihuang group, 20 rats in each group. Model group, Liuweidihuang group six operation storm with L4~5 and L5~6 intervertebral space, randomly selected 1 disc injection of tumor necrosis factor alpha of intervertebral disc degeneration model, and with rongeur the disc the parapophysis mark; the sham operation group by the same surgical exposure of L4~5 and L5~6 intervertebral space, without any treatment after suture; the blank group without any surgical intervention. After model establishment, 3 D, six taste glutinous rehmannia group received 26 mg kg-1 to six Dihuang capsules suspension gavage, the remaining 3 groups with normal saline, 1 times a day for 8 weeks. 2,4,6,8 weeks respectively after drug intervention began from. 5 rabbits were randomly selected from each group The son killed, the model group and the group of Six Ingredient Rehmannia surgery to remove the model of intervertebral disc, control group and sham operation group were randomly selected for L4~5 or L5~6 intervertebral disc removed. Determination of intervertebral disc tissue 1 respectively by polymerase chain reaction method and Western Blot method, mRNA and protein expression of type II collagen level. Results: drug intervention after 2,4,6,8 weeks, compared the expression level of 4 groups of rabbit intervertebral disc tissue collagen mRNA, differences between groups were statistically significant [(0.0648 + 0.0098), (0.0686 + 0.0127), (0.1920 + 0.0406), (0.1246 + 0.0120), F=49.752, P=0.000; (0.0660 + 0.0100), (0.0771 + 0.0118), (0.2524 + 0.0399), (0.1641 + 0.0181), F=79.537, P=0.000; (0.0712 + 0.0112), (0.0891 + 0.0081), (0.3263 + 0.0285), (0.1763 + 0.0150), F=236.726, P=0.000; (0.0941 + 0.0120), (0.1550 + 0.0038), (0.4514 + 0.0396), (0.2058 + 0.0181), F=201.055, P=0.0 00]; the expression of collagen type mRNA in model group was higher than that of blank group, sham operation group and six group (P=0.000, Rehmannia P=0.015, P=0.002, P=0.000; P=0.013, P=0.002, P=0.000, P=0.015; P=0.002, P=0.012, P=0.000, P=0.000). The blank control group and sham operation group of type I collagen mRNA expression level in each time point, there were no significant differences (F=50.563, P=0.132; F=80.352, P=0.634); collagen type mRNA group and six Dihuang group the expression level at each time point, the differences were statistically significant (F=193.635, P=0.000; F= 284.736, P=0.000), the expression levels of collagen mRNA were increased gradually (P=0.004 P=0.002, P=0.000, P=0.000, P=0.003, P=0.001); drug intervention. After 2,4,6,8 weeks, compared the expression level of 4 groups of rabbit intervertebral disc tissue collagen mRNA group, there were significant differences between [(0.0427 + 0.0080), (0.0412 + 0.0058), (0.0116 + 0.0021), (0.0317 卤0.005 6),F=42.696,P=0.000;(0.038 8卤0.004 3),(0.037 3卤0.004 3),(0.011 5卤0.001 8),(0.031 1卤0.003 5),F=108.110,P=0.000;(0.030 3卤0.005 7),(0.025 9卤0.008 3),(0.007 9卤0.002 7),(0.017 2卤0.002 1),F=52.436,P=0.000;(0.029 3卤0.006 9),(0.023 7卤0.004 6),(0.005 3卤0.001 0),(0.014 8卤0.001 8),F=51.375,P=0.000];妯″瀷缁勨叀鍨嬭兌鍘焟RNA琛ㄨ揪姘村钩浣庝簬绌虹櫧缁，

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