麦粒灸对环磷酰胺小鼠骨髓细胞Bcl-2/Bax表达的影响

Effect of wheat grain moxibustion on the expression of Bcl-2/Bax in bone marrow cells of CTX mice

目的 观察麦粒灸对于环磷酰胺(cyclophosphamide,CTX)小鼠骨髓造血功能的影响,从凋亡相关蛋白B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)/Bcl-2关联X蛋白(Bel-2 associated X,BAX)探讨麦粒灸对缓解骨髓抑制、减少细胞凋亡的保护机制。方法 选45只SPF级CD1(ICR)小鼠,随机分为空白组、模型组、麦粒灸组,每组15只。模型组、麦粒灸组腹腔注射CTX 80mg/kg,每日一次,连续三天,建立小鼠白细胞减少症模型;空白组腹腔注射0.2 mL生理盐水。在造模前和治疗期间进行4次外周血白细胞计数,治疗结束后取小鼠股骨和骨髓,制备骨髓病理切片观察,流式细胞术检测细胞凋亡率,免疫组化法检测Bcl-2/Bax蛋白表达量。结果 (1)治疗结束后,麦粒灸组小鼠外周血白细胞水平明显高于空白组和模型组(P <0.01),模型组仍低于空白组(P <0.05);(2)骨髓病理切片苏木精—伊红染色可见麦粒灸组骨髓组织损害程度较模型组有所减轻,骨髓有核细胞增加;(3)麦粒灸组细胞凋亡率较模型组明显下降(P <0.01),但仍高于空白组(P <0.01)。(4)麦粒灸组Bcl-2蛋白含量较模型组升高(P <0.01),Bax蛋白含量较模型组降低(P <0.01)。结论 麦粒灸可升高CTX小鼠外周血白细胞数,缓解骨髓组织的损害程度,通过提高凋亡相关蛋白Bcl-2/Bax值,降低骨髓细胞凋亡率,保护骨髓造血功能,缓解骨髓抑制。

Objective To observe the effect of wheat grain moxibustion on bone marrow hematopoietic function of cyclophosphamide(CTX) mice,and to explore the protective mechanism of wheat grain moxibustion on relieving bone marrow suppression and reducing cell apoptosis from apoptosis-related protein Bcl-2/Bax.Methods 45 SPF CD1(ICR) mice were randomly divided into blank group,model group and wheat grain moxibustion group,with 15 mice in each group.The model group and the wheat grain moxibustion group were intraperitoneally injected with CTX 80 mg/kg,once a day for three consecutive days to establish a mouse leukopenia model.The blank group was intraperitoneally injected with 0.2 mL normal saline.Before modeling and during treatment,peripheral blood white blood cell(WBC) counts were performed 4 times.After treatment,femur and bone marrow of mice were collected for bone marrow pathological section observation.Cell apoptosis rate was detected by flow cytometry,and protein expression level of Bcl-2/Bax was detected by immunohistochemistry.Results(1) The level of WBC in peripheral blood of wheat grain moxibustion group was significantly higher than that of the blank group and model group(P <0.01),while the model group was still lower than the blank group(P <0.05).(2) HE staining of the bone marrow pathological section showed that the bone marrow tissue damage in the wheat grain moxibustion group decreased compared with the model group,and the bone marrow nucleated cells increased.(3) The apoptosis rate of wheat grain moxibustion group decreased significantly compared with the model group(P <0.01),but still higher than that of the blank group(P <0.01).(4) The protein content of Bcl-2 in wheat grain moxibustion group increased compared with the model group(P <0.0.1),and the Bax protein content decreased compared with the model group(P <0.01).Conclusion Wheat grain moxibustion can increase the counts of WBC in peripheral blood of CTX mice,alleviate the damage to bone marrow tissue,reduce the apoptosis rate of bone marrow cells by increasing the value of apoptosis-related protein Bcl-2/Bax,protect the hematopoietic function of bone marrow and relieve bone marrow suppression.