Effect of monomers extracted from Spatholobus suberectus Dunn on proliferation of hematopoietic progenitor cells in marrow-depressed mice / 中国组织工程研究

ABSTRACT

BACKGROUND: Hematogenesis of a body mainly depends on the proliferation of hematopoietic progenitor cell (HPC). Hematopoietic functional impairment will occur when hematopoietic cells are injured by radioactive ray or chemical drug. The proliferation of HPC is the key link of promoting hematogenesis.OBJECTIVE: To study the effects of nine monomers extracted from Spatholobus suberectus Dunn (SSD) on proliferation of HPC in marrow-depressed mice. DESIGN: Randomized controlled trial.SETTING: Department of Pharmacy, General Hospital of Chinese PLA.MATERIALS This experiment was conducted in the Department of Clinical Pharmacology, General Hospital of Chinese PLA from November 2002 to February 2003. Totally 348 healthy Kunming mice, weighing 22-25g, clean grade, of irrespective gender, were selected in this study (certification SCXK-2001-001). The animal experiment was approved by the local ethics committee. SSD was provided by Dispensary of Traditional Chinese Medicine, General Hospital of Chinese PLA; monomers (gallocatechin, formononetin, catechin, pyromucic acid, syringic acid, Demethylvestitol, 1,3,5-benzenetriol, ononin, and epicatechin) were extracted from SSD acetoacetate; TGL-16 centrifuger was made in Shanghai 6th Medical Equipment Factory; CO2 incubator was made in SANYO Company, Japan; MK inverted microscope was provided by OLYMPUS Company, Japan.METHODS: Experimental grouping Mice were randomly divided into 29 groups, including normal group; control group; gallocatechin high-, medium-, low-dose groups; formononetin high-, medium-, low-dose groups; catechin high-, medium-, low-dose groups; pyromucic acid high-, medium-, low-dose groups; syringic acid high-, medium-, low-dose groups; Demethylvestitol high-, medium-, low-dose groups; 1,3,5-benzenetriol high-, medium-, low-dose groups; ononin high-, medium-, low-dose groups; epicatechin high-, medium-, low-dose groups with 12 mice in each group. Experimental intervention All the mice except the mice in normal group had been given total body sublethal dose of irradiation by 60Co γ-ray (215.3 rontgen/min, 4 Gy dose rate, irradiation time of 107.5 seconds). Normal saline was injected intraperitoneally into 8 mice in normal group and control group at the third day after inadiation. Stored solution 2,0.4,0.08g/L of each monomer was intraperitoneally injected into the mice in each monomer high-, medium-, low-dose groups, respectively, at the third day after irradiation. Experimental evaluation Thirty minutes after administration, blood of 8 mice in normal, control group and 12 mice in other groups was collected and normal, control and each dose monomer-containing serums were obtained after centrifugation for 15 minutes, filtering through 0.45μm filter membrane. Then 4 mice in normal and control group were killed to study the effects of nine monomers on proliferation of HPC in marrow-depressed mice by counting erythrocyte colony-forming unit (CFU-E), burst-forming uniterythroid (BFU-E), granulocyte-macrophage colony-forming unit (CFU-GM), and megakaryocyte colony-forming unit (CFU-Meg).MAIN OUTCOME MEASURES: CFU-GM, CFU-E, BFU-E, and CFU-Meg in each group. RESULTS: Totally 348 mice were included in the final analysis. CFU-E The quantity of CFU-E in high-dose of catechin, gallocatechin, syningic acid, and epicatechin groups was significantly higher than that in the control group (P<0.05-0.01) while the quantity of CFU-E in medium-and low-dose of catechin and medium-dose of gallocatechin was also significantly higher than that in the control group (P<0.05). CFU-GM Except pyromucic acid and ononin groups, the amount of CFU-GM in other groups was significantly higher than that in the control group (P<0.01). BFU-E Compared with control group, the amount of BFU-E remarkably increased under the effect of each dose of catechin, gallocatechin, syringic acid and high-, medium-dose of epicatechin (P<0.05). CFU-Meg The amount of CFU-Meg in high-, low-dose syringic acid groups, low-dose gallocatechin groups and each dose group of catechin and epicatechin was significantly higher than that in the control group (P<0.05). Amount of all colonies in the control group was significantly lower than that in the normal group (P<0.01). CONCLUSION: Nine monomers extracted from SSD can promote the proliferation of HPC in bone marrow depressed mice. In particular, the activity of catechin to stimulate proliferation is the strongest.