傣药黑皮跌打乙酸乙酯萃取物的抗炎镇痛作用及急性毒性观察

Anti-inflammatory and antinociceptive effects and acute toxicity of the ethyl acetate fraction of Fissistigma polyanthum

目的观察傣药黑皮跌打90%乙醇提取物(FPEE)乙酸乙酯萃取部位(FPEA)的抗炎镇痛作用及急性毒性。方法抗炎作用观察:ICR小鼠随机分为空白组、模型组、阳性对照组及FPEA低、中、高剂量组,阳性对照组给予地塞米松灌胃,FPEA低、中、高剂量组分别给予FPEA 200、400、800 mg/kg灌胃,空白组、模型组给予等体积生理盐水灌胃,持续给药5 d。除空白组外,各组均通过右后足足跖皮下注射角叉菜胶水溶液致炎,于致炎后1、2、3、4、5、6 h测定各组小鼠右后足足容积,计算足肿胀度及足肿胀抑制率;ELISA法检测致炎6 h后小鼠血清白细胞介素1β(IL-1β)、IL-6及肿瘤坏死因子α(TNF-α)。ICR小鼠随机分为模型组、阳性对照组及FPEA低、中、高剂量组,给药方法同上。采用二甲苯诱导小鼠耳肿胀,计算小鼠耳肿胀度及耳肿胀抑制率。镇痛作用观察:ICR小鼠随机分为模型组、阳性对照组及FPEA低、中、高剂量组,除阳性对照药选用阿司匹林及元胡止痛片外,各组给药方式同上。腹腔注射冰醋酸观察小鼠30 min内扭体次数并计算疼痛抑制率;于末次给药后30、60、90、120 min,将小鼠置于恒温热板上,记录小鼠痛阈值。急性毒性观察:ICR小鼠随机分为空白组、FPEE组、FPEA组,雌雄各半。FPEE组、FPEA组分别采用2000 mg/kg FPEE、FPEA灌胃,空白组给予等体积生理盐水灌胃。记录小鼠14 d内体质量,观察结束后处死小鼠,计算心脏、肝脏、脾脏、肺、肾脏及胸腺器官指数。结果与模型组比较,阳性对照组致炎4、5、6 h后足肿胀度减轻,FPEA中、高剂量组致炎5、6 h后足肿胀度减轻(P均<0.05)。致炎6 h后,FPEA高剂量与阳性对照组肿胀抑制率比较差异无统计学意义(P>0.05)。模型组血清IL-1β、IL-6及TNF-α水平均高于空白组,阳性对照组及FPEA低、中、高剂量组血清IL-6、TNF-α水平低于模型组,阳性对照组及FPEA中、高剂量组血清IL-1β水平低于模型组(P均<0.05)。阳性对照组及FPEA低、中、高剂量组耳肿胀度均小于模型组(P均<0.05),FPEA中、高剂量组耳肿胀抑制率与阳性对照组比较差异无统计学意义(P>0.05)。阳性对照组及FPEA低、中、高剂量组扭体次数均少于模型组,FPEA低、中、高剂量组疼痛抑制率均小于阳性对照组(P均<0.05)。阳性对照组及FPEA低、中、高剂量组末次给药60、90、120 min后痛阈值均高于模型组(P均<0.05)。FPEE、FPEA给药剂量为2000 mg/kg时,14 d内小鼠体质量及心、肝、脾、肺、肾、胸腺器官指数与空白组比较差异均无统计学意义(P均>0.05)。结论FPEA具有较好的抗炎镇痛作用,且不显示急性毒性。

Objective To investigate the anti-inflammatory and antinociceptive effects and acute toxicity of the ethyl acetate fraction(FPEA)from the 95%ethanol extraction of Fissistigma polyanthum(FPEE)in vivo.Methods Observation of anti-inflammatory effects:ICR mice were randomly divided into the blank control group,model group,positive group,and low-,medium-,and high-dose FPEA groups.Mice in low-,medium-,and high-dose FPEA groups were gavaged with 200,400,and 800 mg/kg of FPEA,respectively.Mice in the positive group were gavaged with dexamethasone,while mice in the blank control group and model group were gavaged with equal volume of normal saline.Mice were treated daily for five consecutive days.Except for the blank control group,the right hind paw of each mouse was subcutane ously injected with carrageenan to induce edema.The paw volume was measured at 1,2,3,4,5,and 6 h after the injection,then the edema degree and the inhibition rate of paw edema were calculated.The levels interleukin-1β(IL-1β),IL-6,and tumor necrosis factor-α(TNF-α)in serum of the mice at 6 h after the injection were measured by ELISA.ICR mice were randomly divided into model group,positive group,and low-,medium-,and high-dose FPEA groups,and mice in each group were treated with the same methods as above.The ear edema degree and the inhibition rate of ear edema were calculated using the xylene-induced ear edema model.Observation of antinociceptive effects:ICR mice were randomly divided into the model group,positive group,and low-,medium-,and high-dose FPEA groups.Mice in each group were treated with the same methods as described above,but mice in the positive group were treated with the aspirin and Yuanhu Zhitong tablets.Each mouse was given an intraperitoneal injection of acetic acid,then the number of abdominal writhing was recorded for 30 min and the inhibition rate of writhing was calculated.The latency time was determined at 30,60,90,and 120 min after the drug administration.Observation of acute toxicity:ICR mice were randomly divided into the blank control group,FPEE group,and FPEA group(half male and half female).Mice in the FPEE group and FPEA group were gavaged with FPEE and FPEA at a dose of 2000 mg/kg,respective,while mice in the blank control group were gavaged with equal volume of normal saline.The body weight of mice were recorded within 14 days after intragastric administration.At the end of the observation,all mice were sacrificed,then the organ indexes of heart,liver,spleen,lung,kidney,and thymus were calculated.Results Compared with the model group,the paw edema degree of mice in the positive group were reduced at 4,5,and 6 h after the induction of inflammation,and the paw edema degree of mice in mediumand high-dose FPEA groups were reduced at 5 and 6 h after the induction of inflammation(all P<0.05).At 6 h after the induction of inflammation,there was no significant difference in the inhibition rate of paw edema between the high-dose FPEA and positive groups(P>0.05).The levels of IL-1β,IL-6,and TNF-αin serum of the model group were higher than those of the blank control group,the levels of IL-6 and TNF-αin the positive group and low-,medium-,and high-dose FPEA groups were lower than those of the model group,and the level of IL-1βin the positive group and medium-and highdose FPEA groups were lower than those of the model group(all P<0.05).The ear edema degree were lower in the positive group and low-,medium-,and high-dose FPEA groups than in the model group(all P<0.05),and there was no significant difference in the in the inhibition rate of ear edema between the medium-and high-dose FPEA groups and the positive group(all P>0.05).The writhing times of mice in the positive group and low-,medium-,and high-dose FPEA groups were less than those in the model group,and the inhibition rates of writhing in low-,medium-,and high-dose FPEA groups were less than that of the positive group(all P<0.05).The latency time at 60,90,and 120 min in the positive group and low-,medium-,and high-dose FPEA groups were longer than that of the model group(P<0.05).There was no significant difference in the body weight or organ indexes of heart,liver,spleen,lung,kidney,and thymus between the treated(at the dose of 2000 mg/kg)and the blank control groups(all P>0.05).Conclusion The FPEA exhibited better anti-inflammatory and antinociceptive effects,and did not show acute toxicity.