三七总皂苷通过调节TLR4/TLR2-NF-κB信号通路对再生障碍性贫血小鼠骨髓造血功能的影响

Panax notoginseng saponins regulates hematopoietic function of bone marrow in aplastic anemia mice via TLR4/TLR2-NF-κB pathway

目的探讨三七总皂苷(PNS)对免疫介导再生障碍性贫血(aplastic anemia,AA)小鼠骨髓造血功能和TLR4/TLR2-NF-κB信号通路的调控作用。方法将C57BL/6小鼠随机分成正常组、单纯辐照组(TBI)、模型组、环孢素治疗组和PNS低、中、高剂量治疗组,C57小鼠经5.0 Gy X射线全身辐照和混合淋巴细胞尾静脉输注,建立免疫介导AA小鼠模型,连续给药15 d后测量小鼠体质量,计算脾脏及胸腺指数,检测小鼠骨髓病理、外周血三系细胞、骨髓有核细胞(BMCs)以及外周血清TNF-α、IL-2、IL-10水平,测定骨髓中CD11c及TLR4/TLR2-NF-κB通路相关蛋白表达情况。结果与正常组相比,模型组小鼠体质量、胸腺指数、外周血三系、BMCs及血清IL-10水平明显降低(P<0.05),脾脏指数、结论PNS可改善AA骨髓损伤及免疫紊乱,促进造血,可能与调节骨髓DCs数量及TLR4/TLR2-Akt-NF-κB通路有关。

Aim To explore the effects of Panax notoginseng saponins(PNS)on hematopoietic functions and regulation on the TLR4/TLR2-NF-κB signaling pathway in immune-mediated aplastic anemia(AA)C57 mice.Methods C57BL/6 mice were randomly divided into control group,total body irradiation group(TBI),model group,cyclosporine treatment group,PNS low-dose group,medium-dose group and high-dose group.The immune-mediated AA mice model was established by total body irradiation with 5.0 Gy X-ray and mixed lymphocyte infusion.The body weight was measured,the spleen and thymus index was calculated,bone marrow pathology,the levels of peripheral blood triline cells,bone marrow nucleated cells(BMCs)and the levels of serum TNF-α,IL-2,IL-10 were detected,and the expression of CD11c and proteins related to the TLR4/TLR2-NF-κB pathway were detected 15 days later.Results Compared with control group,body weight,thymus index,the number of peripheral blood triline cells,BMCs and serum IL-10 levels of the mice in model group significantly decreased(P<0.05),while spleen index,the serum TNF-α,IL-2 levels and the protein expression of CD11c,TLR4,TLR2,MyD88,Akt and NF-κB in bone marrow significantly increased(P<0.05).Compared with model group,after PNS treatment,body weight,thymus index,the number of peripheral blood triline cells,BMCs and serum IL-10 levels increased.Spleen index,serum TNF-α,IL-2 levels and the expression of CD11c,TLR4,TLR2,NF-κB and Akt in bone marrow decreased,and the therapeutic effect was not dose-dependent.There was no significant change in the expression of MyD88 and MAPK proteins.Conclusions PNS can improve AA bone marrow injury,regulate immune disoders and promote hematopoiesis,which may be related to the regulation of the number of DCs and the TLR4/TLR2-Akt-NF-κB pathway in bone marrow.