电针对腰椎间盘突出症模型大鼠椎间盘组织环氧合酶1和环氧合酶2 mRNA表达水平的调控

Electroacupuncture for regulating the expressions of cyclooxygenase 1 and cyclooxygenase 2 mRNA in the intervertebral disk tissues of rats with lumbar intervertebral disc protrusion

目的:观察电针治疗对腰椎间盘突出症模型大鼠椎间盘组织环氧合酶基因表达的影响,并与单独给予莫比可及莫比可与电针联合治疗效果进行比较。方法:实验于2004-09/2005-06在上海交通大学附属第六人民医院分子生物学实验室完成。①选用75只清洁级3月龄SD雄性大鼠。取63只大鼠,咬除L5～6棘突、右侧椎板和关节突,暴露椎间盘。用粗针头在纤维环上钻孔,使髓核与硬膜外腔相通建立腰椎间盘突出症模型。②将造模成功大鼠48只按随机数字表法分为4组:电针组、莫比可组、针药组、模型组,每组12只。设其余12只未造模大鼠为正常对照组。电针组:取穴:L4～6夹脊穴(双),环跳(右侧),阳陵泉(右)。电针夹脊穴,深度为0.4cm,疏密波,电流强度4mA,频率2Hz,时间20min,于模型建立后次日开始治疗,1次/d,连续14次。莫比可组:按400mg/kg剂量灌服莫比可药液,1次/d,连续14次。针药组:取穴:L4～6夹脊穴(双),环跳(右侧),阳陵泉(右)。电针夹脊穴,深度0.4cm,疏密波,电流强度4mA,频率2Hz,时间20min;电针治疗结束后根据400mg/kg剂量灌服莫比可药液,于模型建立后次日开始治疗,两种治疗方法均为1次/d,连续14次。模型组:不作任何治疗,灌服等量的生理盐水。正常组:不作任何治疗,灌服等量的生理盐水。③于治疗结束24h后5组同时处死。采用反转录聚合酶链反应的方法检测病变椎间盘组织环氧合酶1以及环氧合酶2mRNA表达。结果:因死亡及造模失败脱失15只,最终进入结果分析大鼠60只。①大鼠椎间盘组织环氧合酶1mRNA表达:模型组为(1.1921±0.3705),低于正常组(6.0541±1.1269,P<0.01);针药组和莫可比组分别为0.4672±0.2813,0.5575±0.1281,均低于电针组和模型组(3.3362±0.8407,P<0.01)。②椎间盘组织环氧合酶2mRNA表达:模型组为4.2494±0.6529,明显高于正常组(0.3116±0.1384,P<0.01);电针组、莫比可组、针药组分别为0.9761±0.7261,1.3330±0.1765,0.6970±0.3193,低于模型组(P<0.01);莫比可组明显高于针药组(P<0.05);正常组与针药组相近(P>0.05)。结论:①单独电针或莫比可治疗及两者联用均可显著抑制大鼠椎间盘组织内的环氧合酶2mRNA表达水平。②单独莫比可治疗及电针或莫比可联用治疗可显著抑制大鼠椎间盘组织内的环氧合酶1mRNA表达水平。

AIM： To observe the effect of electroacupuncture on the gene expression of cyclooxygenase （COX） in the intervertebral disk tissues of rats with lumbar intervertebral disc protrusion （LIDP）, and compare the effect between mobic alone and mobic-electroacupuncture combination. METHODS： The experiment was conducted in the Laboratory of Molecular Biology of the Sixth People＇s Affiliated Hospital of Shanghai Jiaotong University between September 2004 and June 2005. ①Totally 75 male SD rats of 3 months old and cleaning grade were selected. L5-6 processus spinosus, right vertebral plate and processus articularis of 63 rats were removed to expose intervertebral disc. The rat model of lumbar intervertebral disc protrusion was built by puncturing and damaging the annuli fibrosi with bodkin. ②The 48 successful rat models were randomly divided into electroacupuncture group, mobic group, acupuncture plus drug group and model group with 12 in each group. The other 12 rats were considered as normal control group. Rats in the electroacupuncture group were needled at L4-6 Jiaji （Ex-B 2）, Huantiao （GB 30） and Yanglingquan （GB 34） points. The depth of electroacupuncture on Jiaji point was 0.4 cm with current intensity of 4 mA and frequency of 2 Hz for 20 minutes. Therapy was performed at the other day after establishing, once a day for 14 times. Rats in the mobic group were affused in the stomach with mobic （400 mg/kg）, once a day for 14 times. Rats in the acupuncture plus drug group were treated with both the above-mentioned methods in order. After had been treated for 14 days. Rats in the model group and normal group were not treated with any treatment, only perfused with saline of the same volume. ③Twenty-four hours after treatment the rats in the 5 groups were killedl The expressions of COX-1 and COX-2 mRNA in intervertebral disc tissues were tested with reverse transcription-polymerase chain reaction （RT-PCR） method. RESULTS： Fifteen rats were dropped out, because of death or unsuccessful models. A total of 60 rats were involved in the result analysis. ①Expression of COX-1 mRNA in the intervertebral disc tissues of rats： It was lower in the model group （1.1921±0.370 5） than the normal group （6.054 1±1.126 9,P 〈 0.01）. It was lower in the acupuncture plus drug group and mobic group （0.467 2±0.281 3,0.557 5±0.128 1） than the electroacupuncture group and model group （3.336 2±0.840 7 ,P 〈.0.01）. ②Expression of COX-2 mRNA in the intervertebral disc tissues of rats： It was obviouslyhigher in the model group （4.249 4±0.652 9） than the normal group （0.311 6 ±0.138 4,P 〈 0.01）. It was lower in the eleetroaeupuneture g（oup, mobic group and acupuncture plus drug group （0.976 1±0,726 1,1.333 0±0.176 5,0.697 0±0.319 3） than the model group （P 〈 0.01）. It was remarkably higher in the mobie group than the acupuncture plus drug group （P 〈 0.05）. It was similar between the normal group and acupuncture plus drug group （P 〉 0.05）. CONCLUSION： ①Eleetroaeupuneture alone or mobie alone as well as their combination can markedly inhibit the expression of COX-2 mRNA in the intervertebral disc tissues of rats. ② Mobic alone as well as electroaeupuneture and mobie combination can significantly inhibit the expression of COX-1 mRNA in the intervertebral disc tissues of rats.