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四物膏对再生障碍性贫血模型大鼠骨髓造血功能的影响 被引量:7

Effect of Siwu Paste on Hematopoietic Function of Bone Marrow in Rats with Aplastic Anemia and Its Mechanism
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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-κB(NF-κB)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-κB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-κB蛋白及mRNA表达显著升高(P<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-κB蛋白及mRNA表达明显下降(P<0.05,P<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-κB的表达,改善AA模型大鼠骨髓造血功能。 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(ig)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-κB(NF-κB)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-κB protein and mRNA in bone marrow cells were significantly increased(P<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-κB protein and mRNA in bone marrow cells decreased significantly(P<0.05,P<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-κB.
作者 杜青 何丹 黄建华 万丹 苏艳红 唐纯玉 陈林 DU Qing;HE Dan;HUANG Jian-hua;WAN Dan;SU Yan-hong;TANG Chun-yu;CHEN Lin(Hunan Academy of Chinese Medicine,Institute of Chinese Medicine,Changsha 410006,China;Hunan Jiudian Pharmaceutical Co.Ltd.,Changsha 410011,China;Hunan Timesun Pharmaceutical Co.Ltd.,Yongzhou 425006,China)
出处 《中国实验方剂学杂志》 CAS CSCD 北大核心 2021年第3期92-98,共7页 Chinese Journal of Experimental Traditional Medical Formulae
基金 湖南省自然科学基金项目(2019JJ50345,2020JJ5325,2020JJ5328) 湖南省中医药研究院重点项目(201803,201805) 长沙市科技计划项目(kq1901067) 长沙市杰出创新青年培养项目(kq1802017) 中医方证研究转化医学湖南省重点实验室开发基金项目(2019FZ08)。
关键词 四物膏 再生障碍性贫血(AA) 骨髓 Toll样受体4(TLR4) 核转录因子-κB(NF-κB) Siwu paste aplastic anemia bone marrow Toll receptor 4(TLR4) nuclear transcription factor-κB(NF-κB)
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