补肾活血法对慢性再生障碍性贫血小鼠的干预作用研究

doi:10.3877/cma.j.issn.1674-6880.2018.02.002

中华危重症医学杂志(电子版) ›› 2018, Vol. 11 ›› Issue (02) : 78 -82. doi: 10.3877/cma.j.issn.1674-6880.2018.02.002

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论著

补肾活血法对慢性再生障碍性贫血小鼠的干预作用研究

孙菊¹, 刘梅¹, 吴迪炯², 刘文宾², 胡慧瑾², 叶宝东²^,^†(

)

^1. 310053　杭州，浙江中医药大学第一临床医学院
^2. 310006　杭州，浙江中医药大学附属第一医院血液科

收稿日期:2018-02-17 出版日期:2018-04-01
通信作者: 叶宝东
基金资助:
国家自然科学基金资助项目(81373634); 浙江省卫计委创新人才资助计划项目(1S21702); 浙江中医药大学附属第一医院三鹰人才资助计划项目(ZD01601)

Intervention effect of invigorating kidney and activating blood in mice with chronic aplastic anemia

Ju Sun¹, Mei Liu¹, Dijiong Wu², Wenbin Liu², Huijin Hu², Baodong Ye²^,^†()

^1. The First Clinical Medical Institute, Zhejiang Chinese Medical University, Hangzhou 310053, China
^2. Department of Hematology, the First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou 310006, China

Received:2018-02-17 Published:2018-04-01
Corresponding author: Baodong Ye
About author:
Corresponding author: Ye Baodong, Email: 13588453501@163.com

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引用本文：

孙菊, 刘梅, 吴迪炯, 刘文宾, 胡慧瑾, 叶宝东. 补肾活血法对慢性再生障碍性贫血小鼠的干预作用研究[J]. 中华危重症医学杂志(电子版), 2018, 11(02): 78-82.

Ju Sun, Mei Liu, Dijiong Wu, Wenbin Liu, Huijin Hu, Baodong Ye. Intervention effect of invigorating kidney and activating blood in mice with chronic aplastic anemia[J]. Chinese Journal of Critical Care Medicine(Electronic Edition), 2018, 11(02): 78-82.

目的

探讨补肾活血法对慢性再生障碍性贫血小鼠的干预作用及其机制。

方法

建立肾虚合并慢性再生障碍性贫血小鼠模型，将30只小鼠分成中药组、模型组和对照组，每组10只。中药组小鼠给予中药汤剂灌胃0.2 mL/次，2次/d，共60 d，模型组和对照组小鼠给予灌胃相同剂量等渗NaCl溶液。观察小鼠一般情况；于苯试剂皮下注射第16、25次后检测所有小鼠的白细胞、血红蛋白及血小板水平；采用酶联免疫吸附试验检测血管内皮生成因子（VEGF）和基质细胞衍生因子1a（SDF-1a）；同时，电子显微镜下计算骨髓有核细胞个数；苏木素-伊红（HE）染色观察骨髓病理情况。

结果

模型组和中药组的小鼠逐渐出现体质量下降、乏力萎靡、毛发枯疏、四肢苍白等表现。随着中药治疗时间延长，中药组小鼠的上述症状均有不同程度恢复。三组小鼠间白细胞、血红蛋白及血小板计数的比较，差异均有统计学意义（F=16.536、9.273、5.667，P均< 0.05），与对照组相比，模型组及中药组小鼠相同时间点上述指标均明显下降（P均< 0.05），且当苯试剂皮下注射第25次后，中药组小鼠的白细胞、血红蛋白及血小板较模型组均明显升高（P均< 0.05）。三组小鼠间VEGF、SDF-1a水平及骨髓有核细胞的比较，差异均有统计学意义（F=11.231、7.924、8.455，P均< 0.05），且与对照组比较，模型组及中药组小鼠的VEGF[（342 ± 11）、（235 ± 13）、（278 ± 13）ng/L]、SDF-1a[（110 ± 13）、（85 ± 11）、（93 ± 12）ng/L]水平及骨髓有核细胞[（94 ± 15）、（72 ± 11）、（83 ± 13）个/HP]均明显下降，且模型组较中药组更低（P均< 0.05）。HE染色显示中药组及模型组小鼠的粒、红系及巨核细胞均有不同程度减少，但模型组上述情况更严重。

结论

补肾活血祛瘀法能够改善慢性再生障碍性贫血小鼠的临床症状，对其造血恢复有一定的作用，可能通过恢复其VEGF和SDF-1a水平有关。

关键词: 贫血，再生障碍性, 小鼠, 干预性研究, 补肾活血法

Objective

To explore the intervention effect of invigorating kidney and activating blood in mice with chronic aplastic anemia (CAA) and its mechanism.

Methods

The mice model of kidney deficiency complicated with CAA was established. Totally 30 mice were divided into the medicine group, model group and control group, 10 mice in each group. Mice in the medicine group were given orally herbal soup (0.2 mL each time, 2 times a day for 60 days), and mice in the model group and control group only received the same dose of normal saline. The general condition was observed. The levels of white blood cells, hemoglobin and platelets were detected after injection of benzene for 16 and 25 times. The levels of serum vascular endothelial growth factor (VEGF) and stromal cell-derived factor-1a (SDF-1a) were determined by enzyme linked immunosorbant assay. Meanwhile, the bone marrow nucleated cells were counted under electron microscope, and the bone marrow pathological features were observed through hatmatoxylin-eosin (HE) staining.

Results

The mice in the medicine group and model group gradually showed weight loss, fatigue, dry and sparse hair, pale limbs and so on, and as the Chinese medicine intervention prolonged, the mice in the medicine group recovered to varying degrees. The levels of white blood cells, hemoglobin and platelets all showed significant differences among three groups (F=16.536, 9.273, 5.667; all P<0.05). Above indicators at the same time in the model group and medicine group were much lower than those in the control group (all P<0.05), and above indicators after benzene injection for 25 times in the medicine group were much higher than those in the model group (all P<0.05). The levels of VEGF, SDF-1a and bone marrow nucleated cells also showed significant differences among three groups (F=11.231, 7.924, 8.455; all P<0.05). The levels of VEGF [(342 ± 11), (235 ± 13), (278 ± 13) ng/L], SDF-1a [(110 ± 13), (85 ± 11), (93 ± 12) ng/L] and bone marrow nucleated cells [(94 ± 15), (72 ± 11), (83 ± 13)/HP] in the model group and medicine group decreased more obviously compared with the control group, and they decreased most in the model group (all P<0.05). HE staining revealed that granulocyte, erythrocyte and megakaryocyte series decreased in the model group and medicine group, and the model group was less severe.

Conclusion

Invigorating kidney and activating blood may improve mice with CAA through enhancing the levels of serum VEGF and SDF-1a.

Key words: Anemia, aplastic, Mice, Intervention studies, Invigorating kidney and activating blood

表1 各组小鼠苯试剂皮下注射第16、25次后白细胞、血红蛋白及血小板的变化（±s）

组别	次数	例数	白细胞（× 10⁹/L）	血红蛋白（g/L）	血小板（× 10⁹/L）
对照组	16	10	11.2 ± 1.2	171 ± 13	1 119 ± 125
?	25	10	9.8 ± 1.1	165 ± 13	987 ± 146
模型组	16	8	4.6 ± 1.2^a	125 ± 12^ab	890 ± 113^a
?	25	8	2.8 ± 1.1^ab	111 ± 11^ab	719 ± 93^ab
中药组	16	9	5.1 ± 0.9^a	133 ± 15^a	812 ± 114^a
?	25	9	4.8 ± 1.3^a	122 ± 13^a	814 ± 111^a
F值	?	?	16.536	9.273	5.667
P值	?	?	<0.001	0.009	0.035

表1 各组小鼠苯试剂皮下注射第16、25次后白细胞、血红蛋白及血小板的变化（±s）

组别	次数	例数	白细胞（× 10⁹/L）	血红蛋白（g/L）	血小板（× 10⁹/L）
对照组	16	10	11.2 ± 1.2	171 ± 13	1 119 ± 125
?	25	10	9.8 ± 1.1	165 ± 13	987 ± 146
模型组	16	8	4.6 ± 1.2^a	125 ± 12^ab	890 ± 113^a
?	25	8	2.8 ± 1.1^ab	111 ± 11^ab	719 ± 93^ab
中药组	16	9	5.1 ± 0.9^a	133 ± 15^a	812 ± 114^a
?	25	9	4.8 ± 1.3^a	122 ± 13^a	814 ± 111^a
F值	?	?	16.536	9.273	5.667
P值	?	?	<0.001	0.009	0.035

表2 各组小鼠外周血清中VEGF和SDF-1a活性的变化（ng/L，±s）

组别	例数	VEGF	SDF-1a
对照组	10	342 ± 11	110 ± 13
模型组	8	235 ± 13^ab	85 ± 11^ab
中药组	9	278 ± 13^a	93 ± 12^a
F值	?	11.231	7.924
P值	?	0.006	0.014

表2 各组小鼠外周血清中VEGF和SDF-1a活性的变化（ng/L，±s）

组别	例数	VEGF	SDF-1a
对照组	10	342 ± 11	110 ± 13
模型组	8	235 ± 13^ab	85 ± 11^ab
中药组	9	278 ± 13^a	93 ± 12^a
F值	?	11.231	7.924
P值	?	0.006	0.014

图1 各组小鼠骨髓象病理表现

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