不同剂量脑醒喷鼻剂对再灌注损伤脑组织自由基及一氧化氮合酶变化的干预

背景脑醒喷鼻剂由川芎和石菖蒲等中药组成,<神农本草经>谓其"主脑卒中入脑"之功效.目的观察脑醒喷鼻剂对大鼠局灶性脑缺血再灌注损伤脑组织自由基及一氧化氮合酶变化的影响,并与经典西药尼莫地平相比较.设计随机对照实验.单位一所中医药大学医院的内科.对象清洁级成年雄性Wistar大鼠70只,随机分为7组脑醒喷鼻剂高剂量组(高剂组),脑醒喷鼻剂中剂量组(中剂组),脑醒喷鼻剂低剂量组(低剂组),尼莫地平腹腔注射组(尼莫地平组),生理盐水喷鼻剂组(生理盐水组),溶酶喷鼻剂组(溶酶组),假手术组,每组10只.方法除假手术组外,其他6组阻断大鼠左侧大脑中动脉建立大鼠局灶脑缺血再灌注模型.在造模前3 d及再灌注期间,脑醒喷鼻剂高、中、低剂组分别以含川芎嗪和石菖蒲生药5.4,2.7,1.08 mg/(kg·d)和1 35,0.54,0.27g/(kg·d)的剂量滴鼻,3次/d;生理盐水组和溶酶组以生理盐水、溶酶喷鼻剂0.18 mL/(kg·d)滴鼻,3次/d;尼莫地平组用尼莫地平注射液0.8 mg/(kg·d)腹腔注射,2次/d;假手术组按常规饲养至实验取材.用比色法检测丙二醛、超氧化物歧化酶及一氧化氮合酶.主要观察指标①不同剂量脑醒喷鼻剂及其他组动物脑组织丙二醛、超氧化物歧化酶及一氧化氮合酶活性变化.②将不同剂量脑醒喷鼻剂组与尼莫地平组做比较.结果造模时死亡8只动物,进入结果分析62只.①丙二醛含量高剂组、尼莫地平组明显低于生理盐水组(0.92±0.32),(0.87±0.39),(1.35±0.34)μmol/g,P＜0.05],但高剂组和尼莫地平组间无差异.②超氧化物歧化酶活性高剂组、尼莫地平组明显高于生理盐水组(35.64±11.67),(33.88±7.15),(20.70±3.88)NU/mg,P＜0.05],但高剂组和尼莫地平组间无差异.③一氧化氮合酶活性及超氧化物歧化酶活性高剂组、尼莫地平组明显高于生理盐水组(4.64±1.22),(5.00±1.10),(3.08±1.12)mkat/g,P＜0.05],但高剂组和尼莫地平组间无差异.④中、低剂组和溶酶组超氧化物歧化酶及一氧化氮合酶活性有所升高,丙二醛含量有所降低,但与生理盐水组比较无差异.结论脑醒喷鼻剂高剂量组能防止脑缺血缺氧所致的脂质过氧化,使丙二醛生成减少,清除自由基损害,并增加一氧化氮合酶活性,对脑组织的缺血再灌注损伤有保护作用,与尼莫地平的作用相当.

Changes of free radicals and nitric oxide synthase in rat brain following ischemia-reperfusion injury due to different dosage brain-awakening nasal sprayer intervention

BACKGROUND: Brain-awakening nasal sprayer is composed of many herbs,such as Chuanxiong and Shichangpu, which were regarded by "Shennong Bencaojing" as having the function of "preventing stroke in the brain".OBJECTIVE: To observe the changes of free radicals and nitric oxide synthase in rat brain following focal ischemic-reperfusional injury due to brain-awakening nasal sprayer intervention and compare with that due to classical nimodipine.DESIGN: A randomized controlled study.SETTING: Department of internal medicine of a hospital affiliated to a traditional Chinese medical university.MATERIALS: Seventy adult male Wistar rats of clean grade, were randomly divided into seven groups: brain-awakening nasal sprayer of higher dosage group, moderate dosage group, lower dosage group, nimodipine intraperitoneal injection group, physical saline nasal sprayer group, menstruum nasal sprayer group, and sham operation group with 10 rats in each.METHODS: Focal brain ischemia-reperfusion model was established by blocking the left cerebral middle artery in rats of all the groups except sham operation group. Three days before model establishment and during reperfusion, rats were given different dosages of brain-awakening nasal sprayer composed of Chuanxiongqin and Shichangpu of 5.4, 2.7, 1.08 mg/(kg · d) and 1.35, 0. 54, 0.27 g/(kg· d), respectively, three times a day; which was replaced by physical saline and menstruum nasal sprayer of 0. 18 mL/ (kg · d),three times a day in physical saline nasal sprayer group and menstruum nasal sprayer group; rats in nimodipine intraperitoneal injection group received intraperitoneal injection of nimodipine by 0. 8 mg/(kg · d) twice a day. Rats in sham operation group were routinely raised. The content of prodialdehyde, superoxide dismutase and nitric oxide synthase were measured with colorimetric method.MAIN OUTCOME MEASURES: ① The changes of prodialdehyde content, superoxide dismutase and nitric oxide synthase activity in rat brain following focal ischemic-reperfusional injury in groups of different dosage of brain-awakening nasal sprayer and other groups. ② Comparison between different dosage brain-awakening nasal sprayer intervention groups and nimodipine intraperitoneal injection group.RESULTS: Eight rats died during model establishment and the other 62 rats entered the results analysis. ① Content of prodialdehyde: It was significantly lower in higher dosage group and nimodipine intraperitoneal injection group than in physical saline nasal sprayer group (0.92 ± 0. 32), (0. 87 ± 0. 39)vs(1.35 ±0. 34) μmol/g, P ＜ 0.05], but there was no difference between higher dosage group and nimodipine intraperitoneal injection group. ② Activity of superoxide dismutase: It was obviously higher in higher dosage group and nimodipine intraperitoneal injection group than in physical saline nasal sprayer group (35.64 ± 11.67), (33.88 ± 7. 15) vs(20. 70 ± 3.88) NU/mg,P ＜ 0. 05 ], but no difference could be observed between higher dosage group and nimodipine intraperitoneal injection group. ③ Activity of nitric oxide synthase and superoxide dismutase: It was found obviously higher in higher dosage group and nimodipine intraperitoneal injection group than in physical saline nasal sprayer group (4.64 ± 1.22), (5.00 ± 1.10) vs (3.08 ± 1.12) mkat/g, P ＜ 0.05], but no difference could be observed between higher dosage group and nimodipine intraperitoneal injection group.④ The activity of nitric oxide synthase and superoxide dismutase slightly increased while prodialdehyde slightly decreased in moderate dosage group,lower dosage group and menstruum nasal sprayer group, which did not differ significantly from physical saline nasal sprayer group.CONCLUSION: Brain-awakening nasal sprayer intervention exerts multiple effects such as preventing lipo-peroxidation following brain ischemic- reperfusional injury, in addition to suppressing prodialdehyde production, attenuating injury induced by free radicals and increasing nitric oxide synthase activity, thereby playing a similar role to nimodipine in protecting against brain ischemic-reperfusional damage