凉血固元益肾汤对肠型急性辐射损伤大鼠骨髓造血功能的保护作用

Protective Effect of Liangxue Guyuan Yishen Decoction on Bone Marrow Hematopoietic Function in Rats with Intestinal Acute Radiation Injury

目的通过动物实验观察凉血固元益肾汤对肠型急性辐射损伤大鼠骨髓造血机能的保护作用。方法清洁级雄性SD大鼠85只,随机分为空白对照组、模型对照组、阳性药对照组、凉血固元益肾汤中剂量组(中药中剂量组)、凉血固元益肾汤高剂量组(中药高剂量组)。在适应性饲养后,于照射前禁食8 h,除空白对照组外,各组均通过全身一次性^(60)Co-γ射线10 Gy(源皮距3 m)照射制备大鼠急性肠型辐射损伤模型。自照射后第1天至处死前,中药中、高剂量组分别按照10 ml/kg给予含生药2.73 g/ml、5.46 g/ml的凉血固元益肾汤浓缩液灌胃,阳性药对照组按10 ml/kg给予生理盐水配置的0.3 g/ml谷氨酰胺混悬液灌胃,空白对照组和模型对照组按10 ml/kg给予生理盐水灌胃,1次/d。在照射后第3、5、10天,除空白对照组外各组分别随机抽取5只检测白细胞、红细胞(红细胞计数、血红蛋白浓度和红细胞比容)、血小板情况,并剖取一侧股骨进行骨髓病理切片染色观察。结果(1)白细胞:与空白对照组比较,各组在照射后第3、5、10天均明显降低(P<0.01),各治疗组与模型对照组及各治疗组组间比较,差异无统计学意义(P>0.05)。(2)红细胞:照射后第3天,与空白对照组比较,模型对照组红细胞计数、血红蛋白浓度和红细胞比容明显降低(P<0.05,P<0.01),中药高剂量组各指标高于模型对照组及阳性药对照组(P<0.01),甚至其红细胞计数和血红蛋白浓度高于空白对照组(P<0.05);照射后第5天,各组红细胞计数、血红蛋白浓度和红细胞比容比较,差异无统计学意义(P>0.05)。照射后第10天,与空白对照组比较各组红细胞计数、血红蛋白浓度和红细胞比容均出现明显降低(P<0.01),且中药中剂量组红细胞计数和红细胞比容水平均高于模型对照组(P<0.05)。(3)血小板:照射后第3天,各组血小板计数比较,差异无统计学意义(P>0.05),但第5天和第10天各组血小板计数均明显低于空白对照组(P<0.01),且各治疗组与模型对照组及各治疗组组间比较,差异无统计学意义(P>0.05)。骨髓HE染色显示,照射后第3天和第5天,各辐射组骨髓内淋系和巨核系造血细胞数量明显下降,骨髓结构疏松紊乱,其中中药中、高剂量组红系造血细胞相对数量较多;照射后第10天,各照射组红系、粒系和巨核系造血细胞基本消失,但中药高剂量组可见以上三系造血细胞数量增多且有增殖再生的趋势。结论大剂量辐射后短期应用凉血固元益肾汤对^(60)Co-γ射线一次性全身照射10 Gy所致急性肠型辐射损伤大鼠的骨髓造血功能有保护作用,尤对红细胞造血功能更为明显,并可一定程度促进骨髓造血细胞再生。

Objective To observe the protective effect of Liangxue Guyuan Yishen Decoction(LXGYD)on bone marrow hematopoietic function in rats with Gastrointestinal-Acute Radiation injury Syndrome(GI-ARS).Methods Eighty-five male SD rats were randomly divided into blank control group,model control group,positive drug control group,Liangxue Guyuan Yishen Decoction medium dose group(Chinese medicine medium dose group),Liangxue GuyuanYishen Decoction high-dose group(Chinese medicine high-dose group).After adaptive feeding for 3 days,rats were fasted at 8 h before radiation.Except for the Control group,rats in each group were irradiated by whole-body one-time 10 Gy^(60)Co-γray(source dermal distance is 3 m)to prepare the experimental animal model of GI-ARS.From d1 after radiation to the day before death,the M D and H D group were respectively given a concentrated solution of water decocted TCM containing crude drugs at 10 ml/kg,the Glu group was given 0.3 g/ml glutamine suspension at 10 ml/kg,and the Control group and Radiation group were given normal saline at 10 ml/kg.Gavage was once a day.After irradiation on D3,D5 and D10,except for the Control group,5 rats were randomly selected from each group to detect the counts of RBC,WBC and PLT in peripheral blood,and bone marrow pathological sections were stained for observation.Results(1)White blood cells:Compared with the blank control group,each group was significantly reduced on the 3;,5;,and 10;day after irradiation(P<0.01).There were differences between the treatment groups and the model control group and the treatment groups.No statistical significance(P>0.05).(2)Red blood cells:On the 3;day after irradiation,compared with the blank control group,the red blood cell count,hemoglobin concentration and hematocrit of the model control group were significantly reduced(P<0.05,P<0.01),and the indicators of the high-dose Chinese medicine group were higher than those of the model The red blood cell count and hemoglobin concentration of the control group and the positive drug control group(P<0.01)were higher than those of the blank control group(P<0.05);on the 5;day after irradiation,the red blood cell count,hemoglobin concentration and hematocrit of each group were compared,The difference was not statistically significant(P>0.05).On the 10;day after irradiation,compared with the blank control group,the red blood cell count,hemoglobin concentration and hematocrit of each group were significantly reduced(P<0.01),and the red blood cell count and hematocrit level of the Chinese medicine medium-dose group were higher than the model control group(P<0.05).(3)Platelets:On the 3;day after irradiation,there was no significant difference in the platelet counts of each group(P>0.05),but the platelet counts of each group were significantly lower than that of the blank control group on the 5;and 10;day(P<0.01),and there was no statistically significant difference between each treatment group and the model control group and each treatment group(P>0.05).The HE staining of bone marrow showed that on the 3;and 5 th day after irradiation,the number of lymphoid and megakaryotic hematopoietic cells in the bone marrow of each radiation group was significantly reduced,and the bone marrow structure was loose and disordered.Among them,the relative number of erythroid hematopoietic cells in the Chinese medicine and high-dose groups was higher.Many;on the 10 th day after irradiation,the erythroid,granulocytic and megakaryotic hematopoietic cells of each irradiation group basically disappeared,but the number of hematopoietic cells of the above three lines increased and there was a trend of proliferation and regeneration in the high-dose group of traditional Chinese medicine.Conclusions Short-term application of LXGYD after large dose of radiation can protect the hematopoietic function of bone marrow in rats with GI-ARS caused by 10 Gy one-time whole body irradiation by^(60)Co-γrays,and can promote the regeneration and repair of bone marrow hematopoietic cells to a certain extent.