本文關(guān)鍵詞：六味地黃丸對兔椎間盤退變模型椎間盤組織中Ⅰ、Ⅱ型膠原表達的影響　出處：《中醫(yī)正骨》2016年08期 　論文類型：期刊論文

  更多相關(guān)文章： 椎間盤退行性變 六味地黃丸 膠原Ⅰ型 膠原Ⅱ型 兔 動物實驗

【摘要】：目的:觀察六味地黃丸對兔椎間盤退變模型椎間盤組織中Ⅰ、Ⅱ型膠原表達的影響。方法:將80只新西蘭兔隨機分為空白組、假手術(shù)組、模型組和六味地黃組,每組20只。模型組、六味地黃組手術(shù)暴露L4~5和L5~6椎間隙,隨機選擇1個椎間盤注射腫瘤壞死因子α進行椎間盤退變造模,并以咬骨鉗咬除該椎間盤上位椎體橫突進行標(biāo)記;假手術(shù)組經(jīng)相同手術(shù)入路暴露L4~5和L5~6椎間隙,不做任何處理后縫合;空白組不進行任何手術(shù)干預(yù)。造模術(shù)后3 d開始,六味地黃組按26 mg·kg-1以六味地黃膠囊混懸液灌胃,其余3組以等量生理鹽水灌胃,每天1次,共8周。分別于藥物干預(yù)開始后2、4、6、8周,從各組隨機選取5只兔子處死,模型組和六味地黃組手術(shù)取出造模椎間盤,空白組和假手術(shù)組隨機選擇L4~5或L5~6椎間盤取出。分別采用聚合酶鏈?zhǔn)椒磻?yīng)法和Western Blot法測定椎間盤組織中Ⅰ、Ⅱ型膠原的mRNA和蛋白表達水平。結(jié)果:藥物干預(yù)2、4、6、8周后,4組兔子椎間盤組織Ⅰ型膠原mRNA表達水平比較,組間差異均有統(tǒng)計學(xué)意義[(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];模型組Ⅰ型膠原mRNA表達水平高于空白組、假手術(shù)組和六味地黃組(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)�？瞻捉M和假手術(shù)組的Ⅰ型膠原mRNA表達水平各時間點間比較,差異均無統(tǒng)計學(xué)意義(F=50.563,P=0.132;F=80.352,P=0.634);模型組和六味地黃組的Ⅰ型膠原mRNA表達水平各時間點間比較,差異均有統(tǒng)計學(xué)意義(F=193.635,P=0.000;F=284.736,P=0.000),Ⅰ型膠原mRNA表達水平均逐漸增高(P=0.004,P=0.002,P=0.000;P=0.000,P=0.003,P=0.001)。藥物干預(yù)2、4、6、8周后,4組兔子椎間盤組織Ⅱ型膠原mRNA表達水平比較,組間差異均有統(tǒng)計學(xué)意義[(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];模型組Ⅱ型膠原mRNA表達水平低于空白組、假手術(shù)組和六味地黃組(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)�？瞻捉M和假手術(shù)組的Ⅱ型膠原mRNA表達水平各時間點間比較,差異均無統(tǒng)計學(xué)意義(F=70.463,P=0.122;F=90.362,P=0.089);模型組和六味地黃組的Ⅱ型膠原mRNA表達水平各時間點間比較,差異均有統(tǒng)計學(xué)意義(F=110.364,P=0.001;F=86.362,P=0.004),模型組和六味地黃組的Ⅱ型膠原mRNA表達水平均逐漸降低(P=0.002,P=0.005,P=0.003;P=0.000,P=0.001,P=0.000)。藥物干預(yù)2、4、6、8周后,4組兔子椎間盤組織Ⅰ型膠原蛋白表達水平比較,組間差異均有統(tǒng)計學(xué)意義[(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];模型組Ⅰ型膠原蛋白表達水平高于空白組、假手術(shù)組和六味地黃組(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)�？瞻捉M和假手術(shù)組的Ⅰ型膠原蛋白表達水平各時間點間比較,差異均無統(tǒng)計學(xué)意義(F=37.485,P=0.365;F=10.376,P=0.583);模型組和六味地黃組的Ⅰ型膠原蛋白表達水平各時間點間比較,差異均有統(tǒng)計學(xué)意義(F=70.362,P=0.000;F=6.375,P=0.001),模型組和六味地黃組的Ⅰ型膠原蛋白表達水平均逐漸增高(P=0.005,P=0.002,P=0.000;P=0.004,P=0.001,P=0.000)。藥物干預(yù)2、4、6、8周后,4組兔子椎間盤組織Ⅱ型膠原蛋白表達水平比較,組間差異均有統(tǒng)計學(xué)意義[(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];模型組Ⅱ型膠原蛋白表達水平低于空白組、假手術(shù)組和六味地黃組(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)�？瞻捉M和假手術(shù)組的Ⅱ型膠原蛋白表達水平各時間點間比較,差異均無統(tǒng)計學(xué)意義(F=17.364,P=0.092;F=15.573,P=0.175);模型組和六味地黃組的Ⅱ型膠原蛋白表達水平各時間點間比較,差異均有統(tǒng)計學(xué)意義(F=17.753,P=0.001;F=13.674,P=0.000),模型組和六味地黃組的Ⅱ型膠原蛋白表達水平均逐漸降低(P=0.002,P=0.004,P=0.001;P=0.000,P=0.001,P=0.000)。結(jié)論:六味地黃丸能適度下調(diào)兔椎間盤退變模型椎間盤組織中Ⅰ型膠原的表達、上調(diào)Ⅱ型膠原的表達,可在一定程度上延緩椎間盤退變進程。
[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 鹵0.005 6),F=42.696,P=0.000;(0.038 8鹵0.004 3),(0.037 3鹵0.004 3),(0.011 5鹵0.001 8),(0.031 1鹵0.003 5),F=108.110,P=0.000;(0.030 3鹵0.005 7),(0.025 9鹵0.008 3),(0.007 9鹵0.002 7),(0.017 2鹵0.002 1),F=52.436,P=0.000;(0.029 3鹵0.006 9),(0.023 7鹵0.004 6),(0.005 3鹵0.001 0),(0.014 8鹵0.001 8),F=51.375,P=0.000];妯″瀷緇勨叀鍨嬭兌鍘焟RNA琛ㄨ揪姘村鉤浣庝簬絀虹櫧緇，

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