拳参水提取物的镇痛作用

背景:拳参别名草河车,具有清热、镇惊、理湿、消肿的功效,也有报道称其水提取物有抗心律失常作用、中枢抑制作用,其镇痛需进一步研究。目的:观察拳参水提取物的镇痛作用,并与吗啡和氨基比林进行比较。设计:完全随机数字表法,随机对照动物实验。单位:赣南医学院药理教研室。材料:实验于2004-03/05在赣南医学院科研中心实验室完成。①选取健康成年昆明种小鼠150只用于以下4个独立实验。②药品:拳参水提取物由沈阳药科大学植化教研室提供(批号:2003061001);盐酸吗啡注射液(沈阳第一制药厂,批号000305);盐酸纳洛酮注射液(盐侨(湖南)制药有限公司,批号20021109)。方法:①拳参水提取物对醋酸引起小鼠扭体反应的实验:取小鼠40只,随机数字表法分为4组:生理盐水组,拳参水提取物0.10,0.15mg/g组,氨基比林组,每组10只;分别腹腔注射0.02mL/g生理盐水,0.10,0.15mg/g拳参水提取物水溶液,0.10mg/g氨基比林溶液。注药后15min腹腔注射6g/L冰醋酸0.01mL/g,记录15min内各组小鼠扭体反应次数。②拳参水提取物对小鼠热板法致痛作用的实验:取雌性小鼠40只,随机数字表法分为4组:生理盐水组,0.10,0.15mg/g拳参水提取物组,吗啡组,每组10只,分别腹腔注射生理盐水、拳参水提取物(剂量同前)水溶液,0.01mg/g吗啡溶液。用GJ-8402型热板测痛仪,致痛温度调节至(55.0±0.5)℃,以舔后足作为痛反应指标,痛阈潜伏期均不超过60s。给药后15,30,60min各测1次痛觉反应。用热板法测定给药后15,30,60min的痛觉反应。③纳洛酮对吗啡、拳参水提取物对小鼠热板法致痛作用的实验:取雌性小鼠30只,采用随机数字表法分为3组:生理盐水组,纳洛酮 吗啡组,纳洛酮 拳参水提取物组,每组10只,分别腹腔注射0.02mL/g生理盐水,0.004mg/g纳洛酮溶液 0.01mg/g吗啡溶液,0.004mg/g纳洛酮溶液 0.15mg/g拳参水提取物,热板法测定给药后15,30,60min的痛觉反应。④拳参水提取物对电刺激致痛的实验:小鼠40只,随机数字表法分为4组,10只/组,分别腹腔注射0.02mL/g生理盐水,0.10,0.15mg/g拳参水提取物,1g/L吗啡,于给药后20,35,50,70min重复电刺激,电刺激法测定痛觉反应。主要观察指标:①小鼠扭体反应次数。②热板法致小鼠痛觉反应的时间。③电刺激法致小鼠镇痛率。结果:共选取健康成年昆明种小鼠150只用于4个独立实验,全部进入结果分析。①小鼠扭体反应次数:拳参水提取物0.10,0.15mg/g组及氨基比林组小鼠给药后15min扭体反应次数均少于生理盐水组[(15.1±11.1),(8.0±6.5),(6.3±3.2),(54.1±20.2)次,t=3.532～3.681,P<0.01]。②热板法致小鼠痛觉反应的时间:拳参水提取物0.10,0.15mg/g组和吗啡组小鼠给药后15,30,60min痛觉反应的时间均长于生理盐水组(t=2.676～3.683,P<0.05～0.01)。③纳洛酮 拳参水提取物组小鼠给药后各时间点痛觉反应的时间均长于生理盐水组(t=2.676～3.563,P<0.05～0.01),但纳洛酮 吗啡组与生理盐水组相接近(P>0.05)。④电刺激法致小鼠镇痛率:拳参水提取物0.10,0.15mg/g组及吗啡组给药后20,35,50,70min镇痛率均高于生理盐水组(t=3.455～3.634,P<0.01)。结论:①拳参水提取物具有明显的镇痛作用,其镇痛强度与氨基比林、吗啡相当。②阿片受体拮抗剂纳洛酮可拮抗吗啡的镇痛作用,但不能拮抗拳参水提取物的镇痛作用,这表明拳参水提取物的镇痛作用并非通过激动阿片受体而发挥。

Analgesic effect of polygonum bistorta L. water extract

BACKGROUND: Bistort rhizome is also named as caoheche, which is characterized by clearing heat, relieving convulsion, regulating damp and reducing swelling. Additionally, its water extract is characterized by antiarrhythmia and central inhibition; however, analgesia should be studied further.OBJECTIVE: To observe analgesic effect of polygonum bistorta L. Water extract, and compare with morphine and amidazofen.DESIGN: Completely randomized digital table and randomized controlled animal study.SETTING: Department of Pharmacology, Gannan Medical College.MATERIALS: The experiment was carried out in the Laboratory of Scientific Center of Gannan Medical College from March to May 2004. ① A total of 150 healthy adult Kunming mice were used in the 4 independent experiments. ② Medicines: Polygonum bistorta L. Water extract (Department of Phytochemistry, Shenyang Pharmaceutical University; batch number: 2003061001); morphine hydrochloride solution (Shenyang First Pharmaceutical Factory; batch number: 000305); naloxone hydrochloride solution (Yanqiao pharmaceutical Co. Ltd.; batch number: 20021109).METHODS: ① Effect of polygonum bistorta L. Water extract on twisting-body reaction of mice induced by acetic acid: Forty mice were randomly divided into 4 groups: saline group, 0.10 and 0.15 mg/g polygonum bistorta L. Water extract groups and amidazofen group with 10 in each group. All mice were intraperitoneally injected with 0.02 mL/g saline,0.10 and 0.15 mg/g polygonum bistorta L. Water extract solution and 0.10 mg/g amidazofen, respectively. Fifteen minutes later, mice were intraperitoneally injected with 6 g/L 0.01 mL/g acetic acid glacial to record times of twisting-body reaction within 15 minutes. ② Effect of polygonum bistorta L. Water extract on pain response of mice induced by hot-plate test: Forty female mice were randomly divided into 4 groups:saline group, 0.10 and 0.15 polygonum bistorta L. Water extract groups and morphine group with 10 in each group. All mice were intraperitoneally injected with 0.02 mL/g saline, 0.10 and 0.15 mg/g polygonum bistorta L. Water extract solution and 0.01 mg/g morphine solution, respectively. GJ-8402 hot-plate pain response threshold detector was used in this study; pain response temperature was (55.0±0.5) ℃; pain response after licking hindfoot was regarded as reactive marker; latency of pain response threshold was within 60 s. Pain response was measured at 15, 30 and 60 minutes after administration with hot-plate test. ③ Effect of morphine, naloxone and polygonum bistorta L. Water extract on pain response of mice induced by hot-plate test: Thirty female mice were randomly divided into 3 groups: saline group, naloxone+morphine group and naloxone+polygonum bistorta L. Water extract group with 10 in each group. All mice were intraperitoneally injected with 0.02 mL/g saline, 0.004 mg/g naloxone solution+0.01 mg/g morphine solution and 0.004 mg/g naloxone solution+0.15 mg/g polygonum bistorta L. Water extract solution, respectivelu. Pain response was measured at 15, 30 and 60 minutes after administration with hot-plate test. ④ Effect of polygonum bistorta L. Water extract on pain response of mice induced by electric stimulation: Forty mice were randomly divided into 4 groups with 10 in each group. All mice were intraperitoneally injected with 0.02 mL/g saline, 0.10 and 0.15 mg/g polygonum bistorta L. Water extract and 1 g/L morphine, respectively. Pain response was measured at 20, 35, 50 and 70 minutes after administration with electric stimulus method.MAIN OUTCOME MEASURES: ① Times of twisting-body reaction; ②duration of pain response induced by hot-plate test; ③ analgesic rate induced by electric stimulation.RESULTS: All 150 healthy adult Kunming mice were involved in the final analysis. ① Times of twisting-body reaction: At 15 inutes after administration, times of twisting-body reaction were less in 0.10 and 0.15 mg/g polygonum bistorta L. Water extract group and amidazofen group than those in saline group [(15.1±11.1), (8.0±6.5), (6.3±3.2), (54.1±20.2) times, t=3.532-3.681, P ＜ 0.01]. ② Duration of pain response induced by hot-plate test:At 15, 30 and 60 minutes after administration, durations of pain response induced by hot-plate test were longer in 0.10 and 0.15 mg/g polygonum bistorta L. Water extract group and morphine group than those in saline group (t=2.676-3.683, P ＜ 0.05-0.01). ③ Duration of pain response was longer in naloxone + polygonum bistorta L. Water extract group than that in saline group at each time point after administration (t=2.676-3.563, P＜ 0.05-0.01); however, duration in naloxone + morphine group was close to that in saline group (P ＞ 0.05). ④ Analgesic rate induced by electric stimulation: At 20, 35, 50 and 70 minutes after administration, analgesic rate induced by electric stimulation was higher in 0.10 and 0.15 mg/g polygonum bistorta L. Water extract group and morphine group than that in saline group (t=3.455-3.634, P ＜ 0.01).CONCLUSION: ① Polygonum bistorta L. Water extract has the obviously analgesic effect, whose intensity is close to that of amidazofen and morphine. ② Naloxone, an opiate receptor antagonist, can resist analgesic effect of morphine but not that of polygonum bistorta L. Water extract. This suggests that analgesic effect of polygonum bistorta L. Water extract dose not react through exciting opiate receptor.