Effect of Siwu Paste on Hematopoietic Function of Bone Marrow in Rats with Aplastic Anemia and Its Mechanism

Qing DU; Dan HE; Jian-hua HUANG; Dan WAN; Yan-hong SU; Chun-yu TANG; Lin CHEN

doi:10.13422/j.cnki.syfjx.20202405

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Effect of Siwu Paste on Hematopoietic Function of Bone Marrow in Rats with Aplastic Anemia and Its Mechanism

Pharmacology | 更新时间：2021-02-09

- Effect of Siwu Paste on Hematopoietic Function of Bone Marrow in Rats with Aplastic Anemia and Its Mechanism
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 27, Issue 3, Pages: 92-98(2021)
- 作者机构：
  
  1.湖南省中医药研究院中药研究所，长沙 410006
  2.湖南九典制药股份有限公司，长沙 410011
  3.湖南时代阳光药业股份有限公司，湖南永州 425006
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20202405
  CLC： R2-0;R22;R285.5;R289
- Received：25 October 2020，
  
  Published Online：26 November 2020，
  
  Published：05 February 2021
- 稿件说明：
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杜青,何丹,黄建华等.四物膏对再生障碍性贫血模型大鼠骨髓造血功能的影响[J].中国实验方剂学杂志,2021,27(03):92-98.

DU Qing,HE Dan,HUANG Jian-hua,et al.Effect of Siwu Paste on Hematopoietic Function of Bone Marrow in Rats with Aplastic Anemia and Its Mechanism[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(03):92-98.
杜青,何丹,黄建华等.四物膏对再生障碍性贫血模型大鼠骨髓造血功能的影响[J].中国实验方剂学杂志,2021,27(03):92-98. DOI： 10.13422/j.cnki.syfjx.20202405.

DU Qing,HE Dan,HUANG Jian-hua,et al.Effect of Siwu Paste on Hematopoietic Function of Bone Marrow in Rats with Aplastic Anemia and Its Mechanism[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(03):92-98. DOI： 10.13422/j.cnki.syfjx.20202405.

摘要

目的

探讨四物膏对再生障碍性贫血（AA）模型大鼠骨髓造血功能和Toll样受体4（TLR4）/核转录因子-

B（NF-

B）信号通路的调控作用。

方法

SD大鼠随机分为正常组、模型组、阳性药（复方阿胶浆10.8 g·kg

-1

）组、四物膏高、低剂量（22.68，5.67 g·kg

-1

）组，采用乙酰苯肼（APH）联合环磷酰胺（CTX）注射，建立AA大鼠模型，每天灌胃灌胃给予相应药物，连续15 d。检测大鼠外周白细胞（WBC），红细胞（RBC），血红蛋白（HGB），红细胞积压（HCT）和血小板（PLT）水平；计算比较胸腺和脾脏指数；酶联免疫吸附测定（ELISA）检测大鼠血清白细胞介素-3（IL-3）和白细胞介素-6（IL-6）水平，苏木素-伊红（HE）染色观察骨髓病理形态学变化，蛋白免疫印迹法（Western blot）和实时荧光定量聚合酶链式反应（Real-time PCR）检测大鼠股骨骨髓细胞中TLR4和NF-

B蛋白及mRNA表达。

结果

与正常组比较，模型组WBC，RBC，HGB，HCT和PLT水平明显降低，胸腺指数显著下降，脾脏指数显著增加，血清IL-3水平下降，IL-6水平升高，股骨骨髓中粒细胞和巨核细胞减少，髓腔内充填水肿的血管纤维脂肪组织，纤维化明显，骨髓细胞TLR4和NF-

B蛋白及mRNA表达显著升高（

0.01）；与模型组比较，四物膏高剂量组大鼠外周血细胞WBC，RBC，HCT和PLT水平增加，胸腺指数增加，脾脏指数减少，IL-3水平升高，IL-6水平下降，股骨骨髓病理形态轻微改善，骨髓细胞TLR4和NF-

B蛋白及mRNA表达明显下降（

0.05，

0.01）。

结论

四物膏可能通过调控机体骨髓炎症信号通路TLR4/NF-

B的表达，改善AA模型大鼠骨髓造血功能。

Abstract

Objective

To explore the regulatory effect of Siwu paste on the bone marrow hematopoietic function of aplastic anemia （AA） model rats.

Method

SD rats were randomly divided into normal control group， model group， positive drug （Fufang E'jiao Jiang 10.8 g·kg

-1

） group， high-dose Siwu paste （22.68 g·kg

-1

） group and low-dose Siwu paste （5.67 g·kg

-1

） group. Acetophenazine （APH） combined with cyclophosphamide （CTX） injection was used to establish the aplastic anemia rat model. The administration groups were given the corresponding drugs （

） for 15 consecutive days. The levels of white blood cells （WBC）， red blood cells （RBC）， hemoglobin （HGB）， hematocrit （HCT） and platelets （PLT） in peripheral blood cells of rats were detected， thymus and spleen indexes were calculated and compared. Enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of interleukin-3 （IL-3） and interleukin-6 （IL-6） in rat serum. The pathological changes of bone marrow were observed by hematoxylin eosin （HE） staining. Western blot and Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） methods were used to detect Toll receptor 4 （TLR4） and nuclear transcription factor-

B （NF-

B） protein and gene expression in rat femoral bone marrow cells.

Result

Compared with the normal control group， the WBC， RBC， HGB， HCT and PLT levels of the model group were significantly reduced， the thymus index was significantly decreased， the spleen index was significantly increased， the serum IL-3 level was significantly decreased， and the IL-6 level was significantly increased. The number of neutrophils and megakaryocytes in the femoral bone marrow was reduced， and the medullary cavity was filled with edema fibrofatty tissue. The expressions of TLR4 and NF-

B protein and mRNA in bone marrow cells were significantly increased （

0.01）. Compared with the model group， the levels of WBC， RBC， HCT and PLT in peripheral blood cells of rats in the high-dose Siwu paste group increased， the thymus index increased， the spleen index decreased， the IL-3 level was significantly increased， the IL-6 level was significantly decreased， the pathological morphology of femoral bone marrow was slightly improved， and the expressions of TLR4 and NF-

B protein and mRNA in bone marrow cells decreased significantly （

0.05，

0.01）.

Conclusion

Siwu paste may improve the bone marrow hematopoietic function of rats with aplastic anemia by regulating the expression of the bone marrow inflammation signal pathway TLR4/NF-

关键词

Keywords

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