目的 比较胶艾汤不同溶剂提取部位对血虚模型大鼠外周血象、免疫器官指数、白细胞介素2(IL-2)和促红细胞生成素(EPO)水平以及红细胞膜能量代谢酶活性的影响, 筛选胶艾汤补血作用的有效部位。方法 采取每天于眼底静脉丛放血5 mL/kg, 连续放血12 d的方法, 复制大鼠血虚模型;造模同时各组动物分别ig给予胶艾汤及其正丁醇、石油醚、醋酸乙酯、水部位浸膏(剂量均为生药量12 g/kg), 以复方阿胶浆为阳性对照。造模第12天检测各组大鼠外周血红细胞计数(RBC)、血红蛋白(HGB)、红细胞压积(HCT)、血小板计数(PLT), 计算脾脏和胸腺器官指数, 检测血清中IL-2和EPO水平, 检测全血红细胞膜能量代谢酶Na⁺, K⁺-ATP及Ca²⁺, Mg²⁺-ATP酶活性, UPLC法初步分析胶艾汤不同提取部位的化学成分差异。结果 与对照组比较, 模型组大鼠RBC、HGB、HCT、PLT、胸腺指数以及IL-2水平明显降低, Na⁺, K⁺-ATP酶和Ca²⁺, Mg²⁺-ATP酶活性明显减弱, 脾脏指数及EPO水平明显升高, 表明血虚模型复制成功。与模型组比较, 胶艾汤正丁醇组可明显升高HCT、RBC、HGB、PLT(P<0.05、0.01);降低脾脏指数及EPO水平(P<0.05、0.01);明显升高胸腺指数、IL-2水平(P<0.01), 升高红细胞膜Ca²⁺, Mg²⁺-ATP酶及Na⁺, K⁺-ATP酶活性(P<0.05、0.01)。石油醚组可明显升高PLT及IL-2水平(P<0.01), 降低EPO水平及脾脏指数(P<0.05、0.01)。醋酸乙酯组可明显降低脾脏指数及EPO水平(P<0.01), 升高红细胞膜Ca²⁺, Mg²⁺-ATP酶活性(P<0.05)。水部位组可明显升高PLT(P<0.05), 降低EPO水平(P<0.01)。胶艾汤正丁醇提取部位中检测到其他提取部位不含有的咖啡酸、苯甲酰芍药苷、甘草酸铵, 且正丁醇提取部位中没食子酸、原儿茶酸、磷酸川芎嗪、绿原酸的量均高于胶艾汤醋酸乙酯和水提取部位。结论 正丁醇部位是胶艾汤补血作用的最强有效部位。

Objective To screen the active parts with enriching the blood effects of different solvent extraction parts from Jiaoai Decoction (JAD) by comparing the peripheral blood, immune organ index, interleukin 2 (IL-2), and erythropoietin (EPO) and the effect of content of erythrocyte membrane and energy metabolism enzyme activity of blood deficiency model rats. Methods Taking the fundus venous plexus blood 5 mL/kg every day, continuous bleeding for 12 d, to establish the rat model of blood deficiency; The model animals in each group were ig given JAD and n-butanol, petroleum ether, ethyl acetate, parts of water extract (12 g/kg crude drug) with compound E-Jiao slurry as a positive control. On day 12 of modeling, the erythrocyte count (RBC), hemoglobin (HGB), red blood cell hematocrit (HCT), platelet count (PLT) were detected in rats and immune organ indexes of spleen and thymus were calculated, the levels of IL-2 and EPO content were detected, energy metabolism enzymes (Na⁺, K⁺-ATPase and Ca²⁺, Mg²⁺-ATPase) activity were investigated; The difference of the chemical constituents in the different solvent extraction of JAD was analyzed by UPLC. Results Compared with the control group, the RBC, HGB, HCT, PLT, thymus index, and IL-2 levels of rats in the model group decreased significantly, Na⁺, K⁺-ATPase and Ca²⁺, Mg²⁺-ATPase activity weakened significantly, the spleen index and EPO levels were significantly increased, indicating that blood deficiency model was successfully established. Compared with the model group, HCT, RBC, HGB, and PLT (P < 0.05, 0.01) of rats in n-butyl alcohol group could be significantly increased; The spleen index and the content of EPO (P < 0.05, 0.01) were reduced; The thymus index and the content of IL-2 (P < 0.01) were increased, the erythrocyte Ca²⁺, Mg²⁺-ATPase and Na⁺, K⁺-ATPase activity (P < 0.05, 0.01) was elevated. The contents of PLT and IL-2 (P < 0.01) in petroleum ether group could be obviously increased; The content of EPO and the spleen index (P < 0.05, 0.01) were decreased. The ethyl acetate fractions could obviously reduce the spleen index and the content of EPO (P < 0.01), increase Ca²⁺, Mg²⁺-ATPase activity in erythrocytes (P < 0.05). In water group PLT (P < 0.05) could be significantly increased and the content of EPO (P < 0.01) be decreased. There were no coffee acid, benzoyl paeoniflorin, and ammonium glycyrrhizinate in other solvent extracts of JAD, while the content of gallic acid, protocatechuic acid, ligustrazine phosphate, and chlorogenic acid in n-butanol extract were higher than those in ethyl acetate and water extract parts of JAD. Conclusion The n-butanol extract is the most active part of JAD for enriching the blood.

江苏省高校自然科学研究项目(12KJA360002);江苏高校优势学科建设工程资助项目(ysxk-2014)