活血、补肾、化痰三种方法对血管性痴呆大鼠脑组织中超氧化物歧化酶活性和一氧化氮含量的调整

Adjustment of the activity of superoxide dismutase and content of nitric oxide in brain tissues of vascular dementia rats by promoting blood flow,invigorating the kidney and dissipating phlegm methods

目的:观察血管性痴呆大鼠脑组织超氧化物歧化酶活性、一氧化氮含量的变化,比较补肾、活血、化痰三种方法对血管性痴呆大鼠神经毒性的影响,为进一步复方研究和临床用药提供指导和依据。方法:实验于2005-05/09在河北医科大学中医学院实验中心完成。清洁级Wistar雄性健康大鼠53只,采用大鼠脑组织反复缺血再灌注结合腹腔注射硝普钠的方法复制的大鼠拟血管性痴呆模型。造模前将大鼠随机数字法分为6组:假手术组(n=8)、模型组(n=9)、阳性药(尼莫地平)组(n=9)、活血药组(n=9)、补肾药组(n=9)、化痰药组(n=9)。活血药组方为桃仁、红花、当归、川芎、赤芍、丹参各15g。补肾药组方为熟地、山萸肉、枸杞、肉苁蓉、制首乌、菟丝子各15g。化痰药组方为半夏、陈皮、菖蒲、胆南星、枳实、竹茹,各15g。造模前3d开始喂药,造模当天提前2h灌胃,假手术组和模型组给予生理盐水10mL/kg,其余各组分别给予活血药液、补肾药液、化痰药液、尼莫地平液灌胃。每天1次,直至造模7d。造模时将大鼠用10%的水合氯醛麻醉后,分离双侧颈总动脉。模型组、阳性药组及各中草药组在夹闭双侧颈总动脉之前,腹腔注射硝普钠,随即用无创动脉夹夹闭双侧颈总动脉,10min后,再通10min,再夹闭10min。再通后缝合伤口,放回笼中保温饲养。假手术组的麻醉及手术过程与模型组相同,但不阻断颈总动脉、不注射硝普钠。第8天检测大鼠脑组织中超氧化物歧化酶活性、一氧化氮含量。结果:实验大鼠有50只进入结果分析。①血管性痴呆模型组大鼠超氧化物歧化酶活性与假手术组相比明显降低,而用药组大鼠超氧化物歧化酶活性与模型组相比明显升高[假手术组(7.20±0.64)NU/mg,模型组(5.17±0.39)NU/mg,活血药组(6.36±0.55)NU/mg,补肾药组(6.13±0.43)NU/mg,化痰药组(5.99±0.21)NU/mg,P<0.01]。②血管性痴呆模型组大鼠一氧化氮含量与假手术组相比明显升高,而用药组大鼠一氧化氮含量与模型组相比明显降低[假手术组(5.32±0.76)μmol/g,模型组(9.05±0.68)μmol/g,活血药组(6.47±0.25)μmol/g,补肾药组(6.61±0.32)μmol/g,化痰药组(6.85±0.52)μmol/g,P<0.01]。③超氧化物歧化酶活性和一氧化氮含量均有活血药组优于补肾药组的趋势,补肾药组优于化痰药组的趋势,但活血药组、补肾药组、化痰药组之间差异无显著性(P>0.05)。结论:补肾、活血、化痰三种方法均有提高超氧化物歧化酶活性、降低一氧化氮含量的作用,而且呈现出活血法优于补肾法,补肾法优于化痰法的趋势。

AIM： To observe the changes of superoxide dismutase （SOD） and nitric oxide （NO） in brain tissues of rats with vascular dementia, compare the influence of the three methods, invigorating the kidney, promoting blood flow and dissipating phlegm, on the neurovirulence of rats with vascular dementia, provide further direction and basis for complex prescription investigation and clinical medication. METHODS： The experiment was performed at the Experiment Center, Traditional Chinese Medicine College, Hebei Medical University from May to September 2005. Totally 53 clean-grade healthy male Wistar rats were duplicated into vascular dementia models by repeated ischemia/repeffusion in the brain tissue and injected nitroprusside-Na in abdominal cavity. Wister rats were randomly divided into 6 groups before establishing models： sham operation group （n=8）, model group （n=9）, masculine group （n=9）, promoting blood flow group （n=9） and invigorating the kidney group （n=9）, apophlegmatisant group （n=9）. Hemorheologic agent formulas contained peach seed, carthamus tinetorius, angelica root, szechwan lovage rhizome, paeoniae radix, danshen root, each 15 g. Invigorating the kidney formulas contained prepared rhizome of rehmannia, fructus corni, lycium, caulis cistanchis, radix polygoni multiflori preparata, dodder seed, each 15 g. Apophlegmatisant formulas contained pinelliae tuber, pericarpium citri reticulatae, acorus calamus, bile afisaema, fruetus aurantii immaturus, bambusae caulis im taeniam, each 15 g. Intragastric administration was started in three days before operation, lntragastric administration was started two hours in advance that day. Sham operation group and model group were intragastfie administed saline 10 mL/kg, and the others groups were intragastfie administed hemorheologic agent, invigorating the kidney physic liquor, apophlegmatisant, nimodipine in order, once a day for seven days. Double common carotid artery was segregated after paralysing with 10% hydral. Model group, masculine group and every Chinese herbal medicine group were injected with nitroprusside-Na in abdominal cavity before occlusion. Then double common carotid artery was occluded by bulldog clamp, ten minutes later loosed it for ten minutes, and then occluded for ten minutes. Wound was sutured after recanalization, and then put into cage and bred in suitable temperature. Sham operation group was same as model group in paralyzing and operation, but did not occlude double common carotid artery or inject with nitroprusside-Na. At day 8, the activity of SOD and content of NO in brain tissues of rats were detected. RESULTS： Fifty experimental rats were involved in the result analysis. ① The activity of SOD evidently decreased in the model group as compared with the sham operation group, while the activity distinctly increased in the drug groups as compared with the mode[ group [（7.20±0.64） NU/mg in the sham operation group, （5.17±0.39） NU/mg in the model group, （6.36±0.55） NU/mg in the promoting blood flow group, （6.13±0.43） NU/mg invigorating the kidney group, （5.99±0.21） NU/mg apophlegmatisant group,P 〈 0.01]. ②The content of NO markedly increased in the model group as compared with the sham operation group, while the content of No remarkably decreased in the drug groups as compared with the model group [（5.32±0.76）μmol/g in the sham operation group, （9.05±0.68） μmol/g in the model group, （6.47±0.25）μmol/g in the promoting blood flow group, （6.61±0.32）μmol/g in the invigorating the kidney group, （6.85±0.52）μmol/g in the apophlegmatisant group,P 〈 0.01]. ③Promoting blood flow group had an advantage as compared with invigorating the kidney group, and invigorating the kidney group had an advantage as compared with apophtegmatisant group, but without significant difference among three groups in the aetivity of SOD and content of NO（P 〉 0.05）. CONCLUSION： The three methods, invigorating the kidney, promoting blood flow and dissipating phlegm, can elevate the activity of SOD and degrade the content of NO, and the results show the tendencies that promoting blood flow is superior to invigorating the kidney, and invigorating the kidney is superior to dissipating phlegm.