四种中成药对气血双虚模型小鼠血象及免疫水平的影响

Effects of four Chinese medicines on the immunity of mice with deficiency in both Qi and blood

目的:为艾滋病抗病毒疗法所致的骨髓不良反应筛选疗效确切的中成药,观察分析参芪颗粒、复方阿胶浆、贞芪扶正颗粒、复方皂矾丸四种中成药对放血和注射环磷酰胺联合复制的气血双虚模型小鼠血象及免疫水平的影响。方法:实验于2005-08/09在河南中医学院药理实验室完成。①参芪颗粒(江西山高制药有限公司生产,批号040702);复方阿胶浆(山东东阿阿胶股份有限公司生产,批号050446);贞芪扶正颗粒(甘肃扶正药业科技股份有限公司生产,批号040803);复方皂矾丸(陕西郝其军制药有限责任公司生产,批号041014);当归补血口服液(郑州市协和制药厂生产,批号041122);环磷酰胺(上海华联制药有限公司生产,批号050101)。②选取清洁级昆明种小鼠150只,随机数字表法分为15组,10只/组:1～3组分别灌服参芪颗粒混悬液3,2,1g/kg;4～6组分别灌服复方阿胶浆30,20,10mL/kg;7～9组分别灌服贞芪扶正颗粒混悬液15,10,5g/kg;10～12组分别灌服复方皂矾丸混悬液2.4,1.6,0.8g/kg;第13组灌服当归补血口服液10g/kg;剩余2组为空白对照组和模型对照组,分别给于同体积生理盐水10g/kg。各组给药1次/d,连续给药10d。③除空白对照组外,其他各组从给药第1天开始建立气血双虚模型。每只鼠尾部放血0.25mL/10g,然后分别于第2,4,6,8天腹腔注射环磷酰胺80,40,40,40mg/kg。空白对照组同时间点仅腹腔注射等体积生理盐水。末次注射环磷酰胺后2h,眼眶取血,一部分用于血象测定,另一部分离心取血清,测定血细胞比容及血清中巨噬细胞集落刺激因子水平;解剖取胸腺和脾脏,检测胸腺皮质厚度、胸腺淋巴细胞数、脾小结大小、脾脏淋巴细胞数病理学指标的变化。结果:150只小鼠全部进入结果分析,放血和注射环磷酰胺并用可成功建立小鼠气血双虚模型。①与模型对照组比较,参芪颗粒3g/kg组、贞芪扶正颗粒10,5g/kg组、复方皂矾丸1.6g/kg组均可升高气血双虚模型小鼠白细胞水平(t=2.18～2.74,P<0.05),贞芪扶正颗粒15g/kg组作用更为显著(t=2.98,P<0.01);参芪颗粒1g/kg组、复方阿胶浆20mL/kg组、贞芪扶正颗粒15,10g/kg组均可升高红细胞水平(t=2.44～2.69,P<0.05),复方阿胶浆30mL/kg组、贞芪扶正颗粒5g/kg组、复方皂矾丸2.4,1.6g/kg组作用更为显著(t=2.91～3.66,P<0.01);当归补血口服液组、复方阿胶浆20mL/kg组、参芪颗粒3,1g/kg组、贞芪扶正颗粒15,10,5g/kg组均可升高血红蛋白水平(t=2.27～2.85,P<0.05),复方阿胶浆30mL/kg组、复方皂矾丸2.4,1.6g/kg组作用更为显著(t=3.07～4.04,P<0.01);当归补血口服液组、参芪颗粒3,2g/kg组均可升高血小板水平(t=2.20～2.41,P<0.05)。②与模型对照组比较,参芪颗粒2g/kg组、贞芪扶正颗粒5g/kg组均可升高气血双虚模型小鼠血细胞比容(t=2.01～2.62,P<0.05),参芪颗粒1g/kg组、复方阿胶浆30,20,10mL/kg组、贞芪扶正颗粒15,10,5g/kg组、复方皂矾丸2.4,1.6,0.8g/kg组作用更为显著(t=3.18～4.36,P<0.01);参芪颗粒2g/kg组、复方阿胶浆30,20,10mL/kg组、贞芪扶正颗粒15,10,5g/kg组、复方皂矾丸2.4,1.6g/kg组均可显著升高巨噬细胞集落刺激因子水平(t=3.60～6.80,P<0.01)。③与模型对照组比较,当归补血口服液组、参芪颗粒3,2,1g/kg组、复方阿胶浆30,20,10mL/kg组、贞芪扶正颗粒15,10,5g/kg组、复方皂矾丸2.4,1.6,0.8g/kg组均可显著增加气血双虚模型小鼠胸腺皮质厚度(t=3.71～9.34,P<0.01),增大脾小结(t=3.36～11.97,P<0.01),增加脾脏淋巴细胞数(t=4.29～10.44,P<0.01);复方阿胶浆30mL/kg组可明显增加小鼠胸腺淋巴细胞数(t=2.45,P<0.05),当归补血口服液组、参芪颗粒3,2,1g/kg组、复方阿胶浆20,10mL/kg组、贞芪扶正颗粒15,10,5g/kg组、复方皂矾丸2.4g/kg组作用更为显著(t=3.22～8.20,P<0.01)。结论:①四种中成药对气血双虚模型小鼠血红蛋白升高作用相近,以复方阿胶浆和贞芪扶正颗粒对白细胞和红细胞水平升高作用为强,以复方阿胶浆和贞芪扶正颗粒对血小板水平升高作用为优。②四种中成药对气血双虚模型小鼠血细胞比容的影响无差异,以贞芪扶正颗粒和复方皂矾丸对巨噬细胞集落刺激因子水平的升高作用为优。③以参芪颗粒、复方阿胶浆、贞芪扶正颗粒对胸腺皮质厚度和淋巴细胞数的促进作用为优,以参芪颗粒、贞芪扶正颗粒、复方皂矾丸对脾小结和脾脏淋巴细胞数的促进作用为优。

AIM： To select effective Chinese medicine for bone marrow suppression induced by anti-AIDs therapy, and analyze the effects of Shenqi granule, compound Ejiao syrup, Zhenqifuzheng granule and compound melanteritum pill on the hemogram and immunity of mice with deficiency in Qi and blood established by bloodletting and injection of cyclophosphamide. METHODS： The experiment was conducted in the Laboratory of Pharmacology, Henan College of Traditional Chinese Medicine between August and September 2005. ①Shenqi granule （manufactured by Jiangxi Shangao Pharmaceutical Co., Ltd with the batch number of 040702）; compound Ejiao syrup （manufactured by Shandong Dongejiao corporation with the batch number of 050446）; Zhenqifuzheng granule （manufactured by Gansu Fuzheng Pharmaceutical company with the batch number of 040803）; Compound melanteritum pill （manufactured by Shanxi Haoqijun Pharmaceutical Company with the batch number of 041014）; Danggui blood-enriching oral lipid （manufactured by Zhengzhou Xiehe Pharmaceutical Company with the batch number of 041122）; Cyclophosphamide （manufactured by Shanghai Hualian Pharmaceutical Company with the batch number of 050101 ）. ② 150 Kunming clean mice were randomly divided into 15 groups with 10 in each group： Mice in the first 3 groups were drenched with Shenqigranule suspension respectively at 3, 2 and 1 g/kg. Mice in the 4^th-6^th groups were drenched with compound Ejiao syrup respectively at 30, 20 and 10 mL/kg. Mice in the 7^th-9^th groups were drenched with Zhenqifuzheng granule suspension respectively at 15, 10 and 5 g/kg. Mice in the 10^th-12^th groups were drenched with compound melanteritum pill suspension respectively at 2.4, 1.6 and 0.8 g/kg. Mice in the 13^th group were drenched with Dangguiblood-enriching oral liquid at 10 g/kg, while mice in the other 2 groups were taken as the blank control group and model control group and administrated with normal saline at the same dose. Mice in all groups were administrated once a day for 10 continuous days. （~ Mice in all groups except those in＇the blank control group were established into models of deficiency in both Qi and blood since the first day of administration. Blood samples （0.25 mL/10 g） were obtained from the tails of mice, and mice were abdominally injected with cyclophosphamice at 80, 40, 40 and 40 mg/kg respectively at the 2^nd, 4^th, 6^th, and 8^th day. Mice in the blank control group were abdominally injected with normal saline at the same time-points and same dose. Blood was obtained from eye sockets at 2 hours after the last injection of cyclophosphamide, and some of the samples were used for the determination of hemogram, while some were separated of the serum by centrifuge. Haematocrit and the level of serum macrophage colony stimulating factor （M-CSF） were determined. Thoracic gland and spleen were obtained by dissection to inspect the changes in cortical thickness （CT） of spleen, thymic lymphocyte （TL）, size of splenic corpuscle and the leukomonocyte number of spleen. RESULTS： A total of 150 mice were involved in the analysis of results＇. Mouse models of deficiency in both Q/and blood were successfully established by bloodletting and injection of cyclophosphamide. ~ Compared with the model control group, the level of white blood cell in 3 g/kg Shenqigranule group, 10, 5 g/kg Zhenqifuzheng granule group, 1.6 g/kg compound melanteritum pill group were significantly enhanced （t=2.18-2.74, P 〈 0.05）, and the effect was remarkable in 15 g/kg Zhenqifuzheng granule group （t=2.98, P 〈 0.01 ）. The level of erythrocyte increased in 1 g/kg Shenqi granule group, 20 mL/kg compound Ejiao syrup group and 15, 10 g/kg Zhenqifuzheng granule group （t=2.44-2.69, P 〈 0.05）, while the effects were marked in 30 mL/kg compound Ejiao syrup group, 5 g/kg Zhenqifuzheng granule group and 2.4, 1.6 g/kg compound melanteritum pill groups （t=2.91-3.66,P 〈 0.01）. The level of hemoglobin increased in Danggui blood-enriching oral liquid group, 20 mL/kg compound Ejiao syrup group, 3, 1 g/kg Shenqigranule groups and 15, 10, 5 g/kg Zhenqifuzheng granule groups （t=2.27-2.85, P 〈 0.05）, while the effects were significant in 30 mL/kg compound Ejiaosyrup group and 2.4, 1.6 g/kg compound melanteritum pill groups （t=3.07-4.04, P〈 0.01）. The level of blood platelet increased in Danggui blood-enriching oral liquid group and 3, 2 g/kg Shenqigranule groups （t=2.20-2.41, P 〈 0.05）. ②Compared with the model control group, the haematocrit of mice with deficiency＇ in both Qi and blood increased in 2 g/kg Shenqi granule group and 5 g/kg Zhenqifuzheng granule group （t=2.01-2.62, P 〈 0.05）, while the effects were remarkable in 1 g/kg Shenqi granule group, 30, 20, 10 mL/kg compound Ejiao syrup groups, 15, 10, 5 g/kg Zhenqifuzhenggranule groups as well as 2.4, 1.6 and 0.8 g/kg compound melanteritum pill groups （t=3.184.36, P〈 0.01）. The level of establish the qi-blood-deflcient mice model by using Cyclophosphamide, the level of M-CSF was greatly increased in 2 g/kg Shenqigranule group, 30, 20, 10 mL/kg compound Ejiao syrup groups, 15, 10, 5 g/kg Shenqifuzheng granule groups as well as 2.4 and 1.6 g/kg compound melanteritum pill groups （t=3.60-6.80, P 〈 0.01 ）. ③ Compared with the model control group, thymic TC in mice with deficiency in both Qi and blood was significantly increased in Danggui blood-enriching oral liquid group, 3, 2, 1 g/kg Shenqi granule groups, 30, 20, 10 mL/kg compound Ejiao syrup groups, 15, 10, 5 g/kg Shenqifuzheng granule groups as well as 2.4, 1.6 and 0.8 g/kg compound melanteritum pill groups （t=3.71-9.34, P 〈 0.01）, while the splenic corpuscle in those groups were enlarged （t=3.36- 11.97, P 〈 0.01）, and the number of splenic lymphocytes increased （t=4.29-10.44, P 〈 0.01）. Thymic lymphocytes were obviously increased in 30 mL/kg compound Ejiao syrup group （t=2.45,P 〈 0.05）, and the effects in Danggui blood-enriching oral liquid group, 3, 2, 1 g/kg Senqi granule groups, 20, 10 mL/kg compound Ejiao syrup groups, 15, 10, 5 glkg Shenqifuzheng granule groups, and 2.4 g/kg compound melanteritum pill groups （t=3.22-8.20, P 〈 0.01）. CONCLUSION： ① Effects of 4 kinds of Chinese drugs are similar in increasing the hemoglobin of mice with deficiency in both Qi and blood, while compound Ejiao syrup and Zhenqifuzheng granule have remarkable effects in increasing the levels of white blood cells and red blood cells, and they are the best in increasing hemoglobin. ② There are no significant differences in the effects of 4 drugs on haematocrit of mice with deficiency in both Qi and blood, while Zhenqifuzheng granule and compound melanteritum pill are excellent in increasing the level of M-CSF. ③ Shenqi granule, compound Ejiao syrup and Zhenqifuzheng granule have excellent promoting effects on thymic CT and number of lymphocytes, while Shenqi granule, Shenqifuzheng granule and compound melanteritum pill are most effective in increasing splenic corpuscle and the lymphocytes.