黄芪和白芍复方制剂干预痛经模型鼠后血液流变学指标变化

Effect of huangqi and baishao compound preparation on changes of hemorheological indexes of model rats with dysmenorrhea

目的:观察黄芪和白芍复方制剂治疗痛经的效果,分析其药理作用。方法:实验于2002-01/2003-12在河北省医学科学院药物研究室完成。选用雌性SD大鼠110只,雌性昆明种小鼠50只。实验共分3个层次:①实验1:选用雌性大鼠50只,随机分为5组:痛经模型组,田七痛经胶囊组,黄芪和白芍复方制剂高、中、低剂量组,每组10只。实验前大鼠背部皮下注射己烯雌酚4d。在注射己烯雌酚的当天开始给药。田七痛经胶囊组按1.6g/kg剂量灌胃田七痛经胶囊混悬液[主要成分为三七、延胡索、小茴香等,广州敬修堂(药业)股份有限公司生产,批号:01220619,0.4g/粒],黄芪和白芍复方制剂高、中、低剂量组按1.6,0.8,0.4g/kg剂量灌胃黄芪和白芍复方制剂混悬液(由黄芪、白芍、益母草、熟地、当归等9味中药组成,由河北省医学科学院药物研究室研制,批号:200207102,0.5g/粒,胶囊内容物为棕黑色粉末,每克相当于原生药5.74g。实验时将黄芪和白芍复方制剂内容物置于乳钵中研细加水研匀,制成不同浓度的混悬液),痛经模型组灌胃等容积生理盐水。注射己烯雌酚第4天及给药后1h,每只大鼠腹腔注射催产素2U,观察30min内引起扭体反应次,而后用放免法测定血浆β-内啡肽水平。②实验2:将雌性小鼠50只随机分为5组:痛经模型组,田七痛经胶囊组,黄芪和白芍复方制剂高、中、低3个剂量组,每组10只。小鼠皮下注射己烯雌酚混悬液,连续9d,从第10天起,每天上午皮下注射己烯雌酚,下午灌胃给药,田七痛经胶囊组按2.0g/kg剂量灌胃田七痛经胶囊混悬液,黄芪和白芍复方制剂高、中、低剂量组按2.0,1.0,0.5g/kg剂量灌胃黄芪和白芍复方制剂混悬液,痛经模型组灌胃等容积生理盐水。连续3d,于末次给药后30min腹腔注射催产素,记录注射后30min内扭体反应次数。③实验3:将其余健康雌性大鼠60只随机分为6组:对照组[灌胃等容量自来水灌胃20mL/(kg·d)],血瘀模型组[蒸馏水灌胃20mL/(kg·d)],复方丹参片组[灌胃复方丹参片(主要成分为丹参、三七、冰片,石家庄神威药业股份有限公司生产,批号:034070,0.25g/片)混悬液含生药4g/(kg·d)],黄芪和白芍复方制剂高、中、低剂量组[按1.6,0.8,0.4g/kg剂量灌胃黄芪和白芍复方制剂混悬液20mL/(kg·d)],每组10只。每组均连续7d。第8天对照组皮下注射生理盐水,其他各组均皮下注射盐酸肾上腺素(8μg/kg)2次,2次间隔2h,第2次给药后立即将动物置冰水中5min,处置后24h。采用锥板旋转法测定全血黏度和全血还原黏度,利用上述参数计算红细胞指数。④计量资料差异比较采用t检验。结果:复方丹参片组及黄芪和白芍复方制剂高、中、低剂量组进行血液流变学指标检测时采血失败分别为1,1,2,2只,进入结果分析分别为9,9,8,8只,其余各组无脱失值。①实验1:田七痛经胶囊组和黄芪和白芍复方制剂高、中、低剂量组大鼠30min扭体次数明显少于痛经模型组(P<0.05～0.01)。黄芪和白芍复方制剂高剂量组大鼠血浆中β-内啡肽水平明显高于痛经模型组(P<0.05)。②实验2:黄芪和白芍复方制剂对痛经小鼠模型30min扭体次数的影响田七痛经胶囊组和黄芪和白芍复方制剂高、中、低剂量组催产素所致小鼠30min扭体次数明显少于痛经模型组(P<0.05～0.01)。③实验3:黄芪和白芍复方制剂高、中、低剂量组和复方丹参片组全血黏度均明显低于血瘀模型组(P<0.05～0.01);复方丹参片组和黄芪和白芍复方制剂中剂量组红细胞阳性指数明显低于模型组(P<0.01,0.05)。结论:①黄芪和白芍复方制剂对痛经有治疗作用,其作用发挥可能与改善体内内源性疼痛物质β-内啡肽含量有关。②黄芪和白芍复方制剂还可通过改善血液流变学指标,起到活血化瘀作用,从而治疗痛经。

AIM： To evaluate the pharmacological effect of the huangqi and ba/shao capsule on treating dysmenorrhea and its mechanism. METHODS： The experiment was completed at the Department of Pharmacology, Hebei Academy of Medical Sciences from January 2002 to December 2003. Totally 110 female SD rats and 50 female Kunming mice were divided into 3 parts： ① Study Ⅰ ： 50 female SD rats were randomly divided into 5 groups： dysmenorrhea model group, tianqi capsule group, high, middle and low dose of huangqi baishao groups with 10 in each group. Diethylstilbestrol was injected subcutaneously into back of rats 4 days before experiment, meanwhile, rats in tianqi capsule group were perfused with 1.6. g/kg tianqi dysmenorrhea suspension （main components： sanqi, yanhusuo, xiao huixiang, etc.; provided by Guangzhou Jingxiu Limited Company, batch number 01220619, 0.4 g/pill）; rats in high, middle and low dose of huangqi baishao groups were perfused with 1.6, 0.8 and 0.4 g/kg huangqi baishao suspension （huangqi, balshao, yimucao, shudi, danggui, etc.; provided by Pharmaceutical Department of Hebei Academy of Medical Sciences, batch number： 200207102, 0.5 g/capsule, brownblack power, 5.74 g raw drug/g）. The mixture was put into morta to make suspension at various concentrations. Rats in dysmenorrhea model group were perfused with the same volume of saline. 2 U pitocin was injected intravenously 4 days after injection and 1 hour after medication to obverse response times of wrenching body within 30 minutes, and then plasma β- endorphin level was measured with radio-immunity method. ② Study Ⅱ ： 50 female Kunming mice were randomly divided into 5 groups： dysmenorrhea model group, tianqi capsule group, high, middle and low dose of huangqi baishao groups with 10 in each group. Mice were injected with suspension for 9 days. From the 10^th day, mice were injected with diethylstilbestrol in the morning and perfused in the afternoon. Mice in tianqi capsule group were perfused with 2.0 g/kg tianqi dysmenorrhea suspension and mice in high, middle and low dose of huangqi baishao groups were perfused with 2.0, 1.0 and 0.5 g/kg huangqi baishao compound suspension respectively. Rats in dysmenorrhea model group were perfused with the same volume of saline for 3 days. 2U pitocin was injected intravenously to obverse response times of wrenching body within 30 minutes. ③ Study Ⅲ： 60 healthy female rats were randomly divided into 6 groups： control group [perfused with 20 mL/（kg-d） tap-water], blood-stasis model group [perfused with 20 mL/（kg,d） distilled water], compound danshen group [perfused With compound danshen （danshen, sanqi, bingpian）; provided by Shijiazhuang Shenwei Pharmeceutical Limited Company, batch number： 034070, 0.25 g/pill, 4 g/（kg·d） raw materials], high, middle and low dose of huangql baishao groups [perfused with 20 mL/（kg·d） huangqi baishao suspension according to 1.6, 0.8 and 0.4 g/kg doses respectively] with 10 in each group. Rats in each group were perfused for 7 days. On the 8^th day, rats in control group were injected subcutaneously with saline, but in other groups were injected with subcutaneously with 8 μg/kg adrenaline twice a day with the interval of 1 hour, After the second medication, animals were immediately putted into ice water for 5 minutes after 24-hour treatment. Totally blood viscosity and total blood recovery viscosity were measured with cones-plate conbeluting method and index of erythrecyte was calculated on the basis of parameters mentioned above. ④ Measurement data were compared with t test. RESULTS： Totally 1, 1, 2 and 2 animals in compound danshen group, high, middle and low dose of huangqi baishao groups were lost because of failing blood collection respectively, and another 9, 9, 8 and 8 entered the final analysis respectively. There was not lost in other groups. ① Study Ⅰ ： Response times of wrenching body in tianqi capsule group, high, middle and low dose of huangqi baishao groups were lower than those in dysmenorrhea model group within 30 minutes （P 〈 0.05-0.01）. Plasma β-endorphin level in high dose of huangqi baishao group was higher than that in dysmenorrhea model group （P 〈 0.05）. ② Study Ⅱ ： Response times of wrenching body of model mice induced by pitocin in tianqi capsule group, high, middle and low dose of huangqi baishao groups were lower than those in dysmenorrhea model group within 30 minutes （P 〈 0.05-0.01）. ③Study Ⅲ： Total blood viscosity in high, middle and low dose of huangqi baishao groups and compound danshen group was lower than that in blood-stasis group （P 〈 0.05-0.01）. The positive indexes of erythrocyte in compound danshen group and middle dose of huangqi baishao group were markedly lower than those in model group （P 〈 0.01, 0.05）. CONCLUSION： ① Compound huangqi baishao preparation can treat dysmenorrhea, and the mechanism is possibly related with β-endorphin content. ② Compound huangqi baishao preparation can also treat dysmenorrhea through improving hemorheological index by promoting blood circulation and eliminating blood stasis.