熟地黄多糖对气血双虚小鼠全血细胞及血清粒-巨噬细胞集落刺激因子水平的影响

Effect of gross polysaccharide of Radix Rehmanniae Preparata on levels of whole blood cells and serum macrophage colony stimulating factor in mice with both qi and blood deficiency

背景:熟地黄为生地黄的炮制加工品,现代研究多集中在熟地黄粗品的止血、补血、抗氧化、抗衰老、抗炎、益智、降糖等方面,也有对机体免疫功能的影响,但缺少熟地黄活性部位对气血双虚模型动物的影响报道。目的:观察熟地黄多糖对气血双虚小鼠全血细胞及血清粒-巨噬细胞集落刺激因子水平的影响。设计、时间及地点:随机对照,动物血液学分子水平检测,于2002-02/06在河南中医学院动物实验中心完成。材料:清洁级昆明种6周龄雄性小鼠60只,由河南省医学实验动物中心提供。地黄购于河南武陟县药材公司,由本院化学室按药典方法经脱脂、水提、醇沉、除蛋白、透析、洗脱、真空干燥等工艺制得熟地黄多糖。当归补血口服液为郑州市协和制药厂产品。环磷酰胺为上海华联制药有限公司产品。方法:取50只小鼠,每只尾部放血0.5mL,分别于第2,4,6,8天腹腔注射环磷酰胺80,40,40,40mg/kg建立气血双虚模型,随机分为5组,高、中、低剂量熟地黄多糖组分别灌胃400,200,100mg/kg熟地黄多糖水溶液,当归补血口服液组灌胃6.6mL/kg当归补血口服液,模型组仅灌胃同体积生理盐水,1次/d,连续给药10d。剩余10小鼠作为空白对照组,仅腹腔注射同体积生理盐水,尾部不放血。主要观察指标:尾部取血检测全血细胞及血清粒-巨噬细胞集落刺激因子水平的变化。结果:60只小鼠均进入结果分析。与空白对照组比较,模型组全血细胞水平显著降低(t=2.838～13.429,P<0.01),血清粒-巨噬细胞集落刺激因子含量明显降低(t=3.266,P<0.05),表明骨髓造血功能受到抑制,造模成功。与模型组比较,高、中剂量熟地黄多糖组和当归补血口服液组均能显著升高红细胞、白细胞、血小板和血红蛋白水平(t=2.998～12.83,P<0.01);低剂量熟地黄多糖组能明显升高红细胞、白细胞、血小板水平(t=2.779～4.653,P<0.01或0.05)。与模型组比较,高、中剂量熟地黄多糖组和当归补血口服液组可显著提高血清粒-巨噬细胞集落刺激因子含量(t=3.740～6.588,P<0.01);低剂量熟地黄多糖组可明显提高血清粒-巨噬细胞集落刺激因子含量(t=2.662,P<0.05)。结论:熟地黄多糖可促进机体的造血机能,给予高、中剂量时对气血双虚小鼠外周血血细胞状况和血清粒-巨噬细胞集落刺激因子水平均有显著改善作用,以中剂量提升作用最强。

BACKGROUND： Radix Rehmanniae Preparata （RRP） is a processing produce of Radix Rehmanniae. Current studies focus on the function of RRP crude product, such as stop bleeding, enrich the blood, anti-oxidation, anti-aging, anti-inflammation, improving the intelligence and reducing blood sugar. Moreover, the influence on the immunologic function is also reported. However, RRP active site influencing animal models of both qi and blood deficiency is absent. OBJECTIVE： To investigate the effect of gross polysaccharide of RRP （GPRRP） on levels of whole blood cells and serum granulocyte-macrophage colony stimulating factor （GM-CSF） of model mice with both qi and blood deficiency. DESIGN, TIME AND SETTING： The experiments, randomized control design for the determination of molecular levels in hematology, were performed in the Animal Experimental Center of Henan University of Traditional Chinese Medicine from February to June in 2002. MATERIALS： Sixty male Kunming mice of clean grade, aged 6 weeks old were selected from Medical Animal Experimental Center of Henan Province. Radix Rehmanniae was purchased from Henan Provincial Wuzhi Medicinal Factory and prepared into GPRRP according to a series of official methods, including defatting, water extraction, alcohol precipitation, deproteinization, dialysis, elution and vacuum drying. Danggui buxue oral solution was a product of Zhengzhou Union Pharmaceutical Factory. Cyclophosphamide was a product of Shanghai Hualian Pharmaceutical Co., Ltd. METHODS： Fifty mice were used to induce models of both qi and blood deficiency by 0.5 mL blood letting from tails and intravenous injection with 80, 40, 40 and 40 mg/kg cyclophosphamide, respectively. All 50 mice with both qi and blood deficiency were randomly divided into 5 groups： 400, 200 and 100 mg/kg GPRRP groups, danggui buxue oral solution group and model group. Other 10 mice were regarded as blank control group and injected with the same volume of saline. Mice in dosage groups were perfused with 400, 200 and 100 mg/kg GPRRP. Mice in model group were perfused with the same volume of saline, and mice in blank control group were perfused with the same volume of saline once a day for 10 days. MAIN OUTCOME MEASURES： The collected blood samples from tails were used to measure the levels of whole blood cells and serum GM-CSF. RESULTS： Total 60 mice entered the final analysis. Compared with blank control group, the levels of whole blood cells were significantly lower （t=-2.838-13.429, P 〈 0.01 ）, and the content of GM-CSF in serum was also reduced in the model group （t=-3.266, P 〈 0.05）. This indicated an inhibition on the hemopoietic function of bone marrow and successful modeling. Compared with model group, the large- and medium-dose GPRRP solution, together with danggui buxue oral solution group could significantly increase the levels of red blood cells, white blood cells, platelets and hemoglobin in both qi and blood deficient model （t=2.998-12.83, P 〈 0.01）; small dose of GPRRP solution could significantly increase the levels of red blood cells, white blood cells and platelet levels （t=2.779-4.653, P 〈 0.01 or 0.05）. Compared with model group, the large- and medium-dose GPRRP solution, together with danggui buxue oral solution group could significantly increase the levels of GM-CSF （t=-3.740 6.588, P 〈 0.01）, small dose of GPRRP solution also had the similar action （t=-2.662, P 〈 0.05）. CONCLUSION： GPRRP has good function of promoting hematopoiesis in body. Large- and medium-dose GPRRP can significantly improve the levels of peripheral blood cells and serum GM-CSF in model mice with both qi and blood deficiency. The actions enhancing serum GM-CSF level is the strongest by the medium-dose GPRRP.