山茱萸提取物对小鼠造血系统辐射损伤的作用研究

Study of effect in Fructus Corni extract on radiation damage of hematopoietic system in mice

目的研究山茱萸提取物对辐射诱导小鼠造血系统损伤的防护作用。方法选取6~8周龄SPF级雄性C57BL/6J小鼠16只,体重20~22 g,随机选取1只分离股骨,体外培养其骨髓细胞,设置细胞对照组、细胞照射组、0.001 mg/ml山茱萸组、0.01 mg/ml山茱萸组。细胞对照组正常培养,细胞照射组进行照射(1 Gy),0.001 mg/ml山茱萸组、0.01 mg/ml山茱萸组给予相应浓度的山茱萸提取物30 min后照射,18 h后观察四组细胞活力,根据细胞活力选择后续实验的山茱萸提取物浓度。比较细胞对照组、细胞照射组、0.001 mg/ml山茱萸组凋亡程度及活性氧水平。将剩余15只小鼠采用随机区组法分为对照组、照射组、山茱萸组,每组5只。照射组、山茱萸组给予2 Gy全身照射,山茱萸组照射前1 h给予山茱萸提取物DMSO溶液0.2 ml(400 mg/kg),对照组和照射组给予等量含8%DMSO的生理盐水。给药7 d后,检测三组外周血血常规和单侧股骨骨髓单个核细胞(BMNC)、造血祖细胞(HPC)、造血干细胞(HSC)数量及其活性氧生成和DNA(γH2AX)损伤程度。结果细胞照射组细胞活力低于细胞对照组,凋亡细胞比例、活性氧水平高于细胞对照组(P<0.05)。0.01 mg/ml山茱萸组细胞活力与细胞照射组比较,差异无统计学意义(P>0.05)。0.001 mg/ml山茱萸组细胞活力高于细胞照射组,凋亡细胞比例、活性氧水平低于细胞照射组(P<0.05)。照射组白细胞、红细胞、血小板、HPC、HSC水平均低于对照组(P<0.05)。三组BMNC水平比较,差异无统计学意义(P>0.05)。山茱萸组红细胞、HPC水平高于照射组(P<0.05),两组白细胞、血小板、HSC水平比较,差异无统计学意义(P>0.05)。三组BMNC、HPC、HSC中活性氧水平及BMNC中γH2AX水平比较,差异无统计学意义(P>0.05)。照射组HPC、HSC中γH2AX水平高于对照组,山茱萸组HPC、HSC中γH2AX水平低于照射组(P<0.05)。结论山茱萸提取物对造血系统辐射损伤有一定的预防作用。

Objective To study the protective effect of Fructus Corni extract on hematopoietic system damage induced by radiation in mice.Methods Sixteen male SPF level C57BL/6J mice aged 6 to 8 weeks,weighing 20 to 22 g,were selected.One of them was randomly selected to isolate the femur and culture its bone marrow cells in vitro.Cell control group,cell irradiation group,0.001 mg/ml Fructus Corni group,0.01 mg/ml Fructus Corni group were set.The cell control group was cultured normally,the cell irradiation group was irradiated(1 Gy),the 0.001 mg/ml Fructus Corni group and 0.01 mg/ml Fructus Corni group were irradiated with corresponding concentrations of Fructus Corni extract for 30 min,and the cell vitality of the four groups was observed 18 h later,and the concentration of Fructus Corni extract was selected according to the cell vitality.The apoptotic degree and reactive oxygen species levels in cell control group,cell irradiation group,and 0.001 mg/ml Fructus Corni group were compared.The remaining 15 mice were divided into control group,irradiation group,and Fructus Corni group with five mice in each group by randomized block method.The irradiation group and Fructus Corni group were given 2 Gy total body irradiation,the Fructus Corni group was given 0.2 ml(400 mg/kg)DMSO solution of Fructus Corni extract 1 h before irradiation,and the control group and the irradiation group were given the same amount of normal saline containing 8%DMSO.After seven days of administration,routine blood and the number of bone mononuclear nucleated cell(BMNC),hematopoietic progenitor cell(HPC),hematopoietic stem cell(HSC),reactive oxygen species,and DNA(γH2AX)damage were detected in peripheral blood of the three groups.Results The cell vitality in the cell irradiation group was lower than that in the cell control group,and the proportion of apoptotic cells and the level of reactive oxygen species were higher than those in the cell control group(P<0.05).There was no significant difference in cell viability between 0.01 mg/ml Fructus Corni group and cell irradiation group(P>0.05).The cell vitality of 0.001 mg/ml Fructus Corni group was higher than that of cell irradiation group,and the percentage of apoptotic cells and reactive oxygen species level were lower than those of cell irradiation group(P<0.05).The levels of white blood cell,red blood cell,platelet,HPC,and HSC in irradiation group were lower than those in control group(P<0.05).There were no significant differences in BMNC level among the three groups(P>0.05).The levels of red blood cell and HPC in Fructus Corni group were higher than those in irradiation group(P<0.05),but there were no significant differences in the levels of white blood cell,platelet,and HSC between the two groups(P>0.05).There were no significant differences in reactive oxygen species in BMNC,HPC,and HSC of three groups,andγH2AX level in BMNC of three groups(P>0.05).γH2AX levels in HPC and HSC of irradiation group were higher than those in control group,andγH2AX levels in HPC and HSC of Fructus Corni group were lower than those in irradiation group(P<0.05).Conclusion Fructus Corni extract can prevent radiation damage of hematopoietic system.