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[1]王棟,潘浩,朱杭,等.強筋壯骨祛風合劑對髓核致炎大鼠背根神經(jīng)節(jié)中3型酸敏感離子通道的影響[J].中醫(yī)正骨,2017,29(01):18-22,26.
　WANG Dong,PAN Hao,ZHU Hang,et al.Effect of Qiangjin Zhuanggu Qufeng Heji on acid-sensing ion channel 3 of dorsal root ganglia in rats with inflammation caused by nucleus pulposus[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2017,29(01):18-22,26.
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強筋壯骨祛風合劑對髓核致炎大鼠背根神經(jīng)節(jié)中3型酸敏感離子通道的影響()
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《中醫(yī)正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第29卷
期數(shù):
2017年01期
頁碼:
18-22,26
欄目:
基礎(chǔ)研究
出版日期:
2017-01-20

文章信息/Info

Title:
Effect of Qiangjin Zhuanggu Qufeng Heji on acid-sensing ion channel 3 of dorsal root ganglia in rats with inflammation caused by nucleus pulposus
作者:
王棟; 潘浩; 朱杭; 諸力; 何永江; 王堅; 賈高永
浙江省杭州市中醫(yī)院,浙江 杭州 310007
Author(s):
WANG Dong; PAN Hao; ZHU Hang; ZHU Li; HE Yongjiang; WANG Jian; JIA Gaoyong
Hangzhou Hospital of Traditional Chinese Medicine,Hangzhou 310007,Zhejiang,China
關(guān)鍵詞:
腰痛;  強筋壯骨祛風合劑;  3型酸敏感離子通道;  神經(jīng)節(jié); 脊;  大鼠; Sprague-Dawley;  動物實驗
Keywords:
low back pain;  Qiangjin Zhuanggu Qufeng Heji;  acid-sensing ion channel 3;  ganglia; spinal;  rats; Sprague-Dawley;  animal experimentation
摘要:
目的:觀察強筋壯骨祛風合劑對髓核致炎大鼠背根神經(jīng)節(jié)中3型酸敏感離子通道(acid-sensing ion channel 3,ASIC3)的影響。方法:選取40只清潔級成年雄性SD大鼠,隨機分為空白組、假手術(shù)組、模型組、阿米洛利組和中藥組,每組8只。除空白組外,其余各組大鼠均手術(shù)暴露L5背根神經(jīng)節(jié)。假手術(shù)組暴露L5背根神經(jīng)節(jié)后直接縫合切口; 模型組和中藥組取出大鼠尾部的髓核組織置于暴露的L5背根神經(jīng)節(jié)上,縫合切口; 阿米洛利組將取自大鼠尾部的髓核組織置于暴露的L5背根神經(jīng)節(jié)上,并在暴露的L5背根神經(jīng)節(jié)周圍滴入含0.1 mg鹽酸的阿米洛利藥液1 mL,縫合切口。自造模后第1天開始,中藥組按5 mL·kg-1以強筋壯骨祛風合劑灌胃,每天2次,連續(xù)30 d; 空白組、假手術(shù)組、模型組不進行藥物干預(yù)。分別于造模前1 d及造模后3、5、9、15、23、29 d,采用Up-Down法測定大鼠左后足50%機械刺激縮足閾值(50% paw withdrawal threshold,50%PWT)。中藥組藥物干預(yù)結(jié)束后,處死所有大鼠,取出L5背根神經(jīng)節(jié)分成2份,一份采用免疫組織化學(xué)法計算ASIC3陽性面積,另一份采用Western Blot技術(shù)檢測ASIC3蛋白表達量。結(jié)果:造模后3 d內(nèi),模型組、阿米洛利組和中藥組均有部分大鼠出現(xiàn)左后足外翻以及足趾背伸現(xiàn)象。造模前后不同時間50%PWT的差異有統(tǒng)計學(xué)意義,即存在時間效應(yīng)(F=50.132,P=0.000)。5組50%PWT總體上比較,組間差異有統(tǒng)計學(xué)意義,即存在分組效應(yīng)(F=288.219,P=0.000)。除造模前1 d時外(F=0.332,P=0.854),造模后各時點5組大鼠50%PWT比較,組間差異均有統(tǒng)計學(xué)意義(F=288.015,P=0.000; F=308.588,P=0.000; F=374.945,P=0.000; F=560.713,P=0.000; F=343.043,P=0.000; F=459.914,P=0.000); 造模后各時點模型組的50%PWT均低于空白組、假手術(shù)組、阿米洛利組和中藥組(P=0.010,P=0.011,P=0.001,P=0.001; P=0.023,P=0.022,P=0.000,P=0.004; P=0.013,P=0.013,P=0.003,P=0.001; P=0.012,P=0.002,P=0.002,P=0.002; P=0.004,P=0.000,P=0.000,P=0.000; P=0.003,P=0.005,P=0.003,P=0.013); 造模后各時點空白組和假手術(shù)組50%PWT比較,差異均無統(tǒng)計學(xué)意義(P=0.640,P=0.710,P=0.590,P=0.480,P=0.330,P=0.390); 造模后各時點阿米洛利組50%PWT均高于中藥組(P=0.021,P=0.011,P=0.013,P=0.002,P=0.001,P=0.005)。時間因素與分組因素存在交互效應(yīng)(F=2.358,P=0.002)。5組大鼠背根神經(jīng)節(jié)中ASIC3陽性面積和ASIC3蛋白表達量比較,組間差異均有統(tǒng)計學(xué)意義(F=205.890,P=0.030; F=101.740,P=0.001); 空白組、假手術(shù)組、阿米洛利組和中藥組背根神經(jīng)節(jié)中ASIC3陽性面積和ASIC3蛋白表達量均小于模型組(P=0.004,P=0.003,P=0.001,P=0.007; P=0.000,P=0.000,P=0.000,P=0.000); 阿米洛利組背根神經(jīng)節(jié)中ASIC3陽性面積和ASIC3蛋白表達量均大于中藥組(P=0.001; P=0.000)。結(jié)論:強筋壯骨祛風合劑可以降低髓核致炎大鼠病變的背根神經(jīng)節(jié)中ASIC3的水平,提高大鼠的疼痛閾值,但其作用效果不及鹽酸阿米洛利。
Abstract:
Objective:To observe the effect of Qiangjin Zhuanggu Qufeng Heji(強筋壯骨祛風合劑,QJZGQFHJ)on acid-sensing ion channel 3(ASIC3)of dorsal root ganglia(DRG)in rats with inflammation caused by nucleus pulposus.Methods:Forty clean adult male SD rats were selected and randomly divided into blank group,sham-operated group,model group,Amiloride group and TCD group,8 cases in each group.The surgeries were performed on rats in sham-operated group,model group,Amiloride group and TCD group to expose L5 DRG.The surgical incisions were sutured in sham-operated group right after the L5 DRG were exposed.The nucleus pulposuses in tails were fetched out and were placed on the denudate L5 DRG in model group and TCD group,and then the surgical incisions were sutured.The nucleus pulposuses in tails were fetched out and were placed on the denudate L5 DRG in Amiloride group,and Amiloride solution(1 ml),containing hydrochloric acid(0.1 mg),was dropped arround the denudate L5 DRG,then the surgical incisions were sutured.Since the 1st day after modeling,the rats in TCD group were intragastric administrated with QJZGQFHJ(5ml/kg),twice a day for 30 consecutive days.No drug interventions were performed on rats in blank group,sham-operated group and model group.The 50% paw withdrawal threshold(50%PWT)of left posterior feet were measured by using Up-Down methods at 1 day before modeling and at 3,5,9,15,23 and 29 days after modeing respectively.After the end of drug interventions,all rats in TCD group were executed and the L5 DRGs were fetched out and were divide into 2 parts.One of them was used for calculating the ASIC3 positive areas by using immunohistochemical method,and the other potion was used for detecting the ASIC3 protein expression by using Western Blot assay.Results:Left posterior foot eversion and toe dorsal extension were found in some rats of model group,Amiloride group and TCD group within 3 days after modeling.There was statistical difference in 50% PWT between different timepoints before and after modeling,in other words,there was time effect(F=50.132,P=0.000).There was statistical difference in 50% PWT between the 5 groups in general,in other words,there was group effect(F=288.219,P=0.000).Except at 1 day before modeling(F=0.332,P=0.854),there was statistical difference in 50% PWT between the 5 groups(F=288.015,P=0.000; F=308.588,P=0.000; F=374.945,P=0.000; F=560.713,P=0.000; F=343.043,P=0.000; F=459.914,P=0.000).At different timepoints after modeling,the 50% PWT were lower in model group compared to blank group,sham-operated group,Amiloride group and TCD group(P=0.010,P=0.011,P=0.001,P=0.001; P=0.023,P=0.022,P=0.000,P=0.004; P=0.013,P=0.013,P=0.003,P=0.001; P=0.012,P=0.002,P=0.002,P=0.002; P=0.004,P=0.000,P=0.000,P=0.000; P=0.003,P=0.005,P=0.003,P=0.013).There was no statistical difference in 50%PWT between blank group and sham-operated group(P=0.640,P=0.710,P=0.590,P=0.480,P=0.330,P=0.390)and the 50% PWT were higher in Amiloride group compared to TCD group(P=0.021,P=0.011,P=0.013,P=0.002,P=0.001,P=0.005)at different timepoints after modeling.There was interaction between time factor and group factor(F=2.358,P=0.002).There was statistical difference in ASIC3 positive areas and ASIC3 protein expression in DRG between the 5 groups(F=205.890,P=0.030; F=101.740,P=0.001).ASIC3 positive areas and ASIC3 protein expression were less in blank group,sham-operated group,Amiloride group and TCD group compared to model group(P=0.004,P=0.003,P=0.001,P=0.007; P=0.000,P=0.000,P=0.000,P=0.000),and were greater in Amiloride group compared to TCD group(P=0.001; P=0.000).Conclusion:QJZGQFHJ can reduce the ASIC3 level in diseased DRG and increase the pain threshold in rats with inflammation caused by nucleus pulposus,while it is inferior to Amiloride in effectiveness.

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備注/Memo

備注/Memo:
基金項目:浙江省中醫(yī)藥科技計劃項目(2015ZA157); 浙江省杭州市科技計劃重點專病專科項目(20150733Q60)
通訊作者:潘浩 E-mail:[email protected]
更新日期/Last Update: 2017-01-20