BACKGROUND: Chinese herb digan oral liquid promotes manufacture of hematopoietic cells. During irradiative injury, how are the therapeutic effects of intervention with Chinese herbal compound on expressions of stromal cellular adherent molecule (SCAM) and cyclin gene that are closely related to hematopoietic cells?OBJECTIVE: Mouse model with irradiative injury was established to assay the effects of Chinese herbal compound-digan oral lipid on the expression of medullary cyclinD1mRNA and the expressions of hematopoietic stem cells (HSCs), stromal cellular adherent molecular CD45 (SCAMCD45) and vascular cellular adherent moldecular-1 (VCAM-1).DESIGN: Randomized controlled experiment was designed.SETTING: Department of Integrated Traditional Chinese and Western Medicine, Wuhan General Hospital of Guangzhou Military Area Command of Chinese PLA; Institute of Hematology of Tongji Medical College and Institute of Hepatology of Hubei College of Traditional Chinese Medicine.MATERIALS: The experiment was performed in Institute of Hematology of Tongji Medical College and Institute of Hepatology of Hubei College of Traditional Chinese Medicine from January 2001 to June 2002, in which,48 outbred Kunming mice were employed, aged varied from 8 to 12weeks, mass-weighted varied from 18 to 22 g, of either sex. Drugs: 100% digan oral lipid is composed of shudihuang (radix rehmanniae preparata) and zhigancao (radix glycyrrhizae preparata). At ratio of 2/1, water decoction was prepared and concentrated as 100% oral liquid (containing raw herb 1 g/mL).rnMETHODS: Totally 48 mice were randomized into 4 groups, 12 mice in each one. Normal group: no any management. Oral lipid group: Digan oral lipid was applied for gastric perfusion in model with irradiative injury,0.02 ml/g, twice a day. Model group: After modeling, physiological saline was used for gastric perfusion with the same dosage as the above. Batilol group: After modeling, batilol suspension was used for gastric perfusion (0.1 g/kg). Gastric perfusion was done continuously for 9 days in each group. On the 10th day, hybridization in situ was used to assay cyclinD1 mRNA expression of medullary mononuclear cell (MNC), Immuniohisto chemical SABC method was used to assay medullary VCAM-1 expression and flow cytometry (FCM) was used to assay CD45 content.MAIN OUTCOME MEASURES: CyclinD1 mRNA expression of medullary mononuclear cell (MNC) and expression of medullary stromal cellular VCAM-1 and CD45 antigenRESULTS: After modeling, there were 12 mice in normal group and 10 mice in rest groups. In result analysis, 5 mice were randomized in each group. ① In oral liquid group, the expression of medullary cellular cyclinD1 mRNA and VCAM-1 were higher significantly than model group[oralliquid group: (27.34±4.49), (53.94±8.67)% and model group: (16.06±4.57),(31.92±5.01)%, P＜ 0.01]. ② Expression of CD45 antigen iroral liquid group was higher significantly than model group [(41.38±5.76), (19.24±4.98)%, P ＜ 0.01]. Both digan oral liquid and batilol improved the expressions of cyclinD1,VCAM-1 and CD45 antigen.CONCLUSION: Chinese herbal compound-digan oral liquid enhances the expression of adherent molecule of hematopoietic stem cells, promotes the expression of stroma] cellular adherent molecule and improves the transcription of medullary cellular cyclinD1, which induces hematopoietic cells to be differentiated more maturely and to be transported peripherally so as to supplement blood.%背景:中药地甘口服液能促进造血细胞的生成,辐射损伤时,与造血细胞密切相关的基质细胞黏附分子和周期素基因表达及中药复方的干预效果如何?目的:建立辐射损伤小鼠模型,并检测中药复方-地甘口服液对该模型小鼠骨髓细胞周期素基因CyclinD1mRNA的表达,造血干细胞和基质细胞黏附分子CD45、血管细胞黏附分子-1表达的作用.设计:随机对照实验.单位:广州军区武汉总医院中西医结合科;同济医学院血液病研究所;湖北中医学院肝病研究所.材料:实验于2001-01/2002-06在同济医学院血液病研究所和湖北中医学院肝病研究所完成.正常8～12周远交系昆明小鼠48只,体质量18～22 g,雌雄兼用.药物:100%地甘口服液由大量熟地黄、炙甘草为主组成,按2:1的比例称质量,经水煎、浓缩配成100%浓度(含生药1g/mL)的口服液备用.方法:48只小鼠随机分为4组,每组12只.正常组:不作任何处理;地甘口服液组:造成辐射损伤模型后用地甘口服液灌胃,0.02 mL/g,2次/d;模型组:按上方法制模后,即用生理盐水灌胃,用量及方法同上;鲨肝醇组:按上方法制模后,用鲨肝醇混悬液灌胃(0.1g/kg),各组均连续灌胃9 d.第10天用原位杂交法检测小鼠骨髓单个核细胞CyclinD1mRNA的表达,用免疫组化SABC法检测骨髓血管细胞黏附分子-1的表达,用流式细胞仪检测CD45的含量.主要观察指标:骨髓单核细胞周期素基因CyclinD1mRNA表达,骨髓基质细胞黏附分子血管细胞黏附分子-1和CD45抗原的表达.结果:造模完成后,正常组12只,余下组各10只,结果分析时各组随机选5只.①地甘口服液组小鼠骨髓细胞CyclinD1mRNA和血管细胞黏附分子-1的表达显著高于模型组[地甘口服液组:(27.34±4.49),(53.94±8.67)%,模型组:(16.06±4.57),(31.92±5.01)%,P＜0.01].②CD45抗原的表达地甘口服液组显著高于模型组[(41.38±5.76),(19.24±4.98)%,P＜0.01],地甘口服液和鲨肝醇均能提高CyclinD1血管细胞黏附分子-1和CD45抗原的表达.结论:中药复方-地甘口服液能增强造血干细胞黏附分子的表达,还能促进基质细胞黏附分子的表达,提高骨髓细胞周期素基因cyclinD1的转录水平,使造血细胞不断的分化成熟和向外周循环输送,起到了补血的作用.
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