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作 者:朱小勤[1] 赵琳[1] 邱仲川[1] 陈珮[1] 黄中迪[1] 胡晓莹[1] 瞿玮颖 陆颖佳[1] 封舟[1] ZHU Xiaoqin;ZHAO Lin;QIU Zhongchuan;CHEN Pei;HUANG Zhongdi;HU Xiaoying;QU Weiying;LU Yingjia;FENG Zhou(Department of Hematology, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 200021, China)
机构地区:[1]上海中医药大学附属曙光医院血液科,上海200021
出 处:《上海中医药杂志》2018年第4期96-100,共5页Shanghai Journal of Traditional Chinese Medicine
基 金:国家自然科学基金青年基金项目(81403233)
摘 要:目的基于P38MAPK途径探讨复方参鹿颗粒含药血清对TNF-α诱导的正常骨髓CD34+细胞凋亡的影响。方法 Wistar大鼠随机分为空白组和复方参鹿颗粒低、中、高剂量组,分别制备空白血清和不同浓度的含药血清。收集并分离8例正常的骨髓单个核细胞(BMMC),分别加入空白血清和10%不同浓度含药血清,与20 ng/ml人重组TNF-α共同培养。流式细胞术检测CD34^+细胞凋亡率,Western blot检测骨髓单个核细胞P38、p-P38、P53、p-P53、Bcl-xl、Bcl-2蛋白表达,荧光定量PCR检测P38、P53、Bcl-xl、Bcl-2mRNA表达。结果与空白组相比,复方参鹿颗粒中、高剂量组CD34^+细胞凋亡率以及p-P38、p-P53蛋白表达显著下降,Bcl-xl蛋白和mRNA表达显著升高,差异有统计学意义(P<0.05)。结论复方参鹿颗粒含药血清可有效抑制TNF-α诱导的正常骨髓CD34^+细胞凋亡,下调p-P38和p-P53蛋白表达,上调Bcl-xl表达。其机制可能与P38MAPK通路介导的凋亡途径有关。Objective To discuss the effects of medicated serum of Compound Shenlu Granule on TNF-α induced CD34+cell apoptosis in normal bone marrow based on P38 MAPK pathway. Methods Wistar rats were randomly divided into the blank group and Compound Shenlu Granule groups with low-,medium-and high-dose. The blank serum and medicated serum at different concentrations were prepared respectively. The normal bone marrow mononuclear cells( BMMCs)from 8 individuals were collected and isolated. The BMMCs were treated with blank serum or 10% medicated serum at different concentrations,which co-cultured with 20 ng/ml TNF-α. The apoptosis rate of CD34^+ cells was detected by flow cytometry. The protein expressions of P38,p-P38,P53,p-P53,Bcl-xl and Bcl-2 in BMMCs were detected by Western blot. The mRNA expressions of P38,P53,Bcl-xl and Bcl-2 were detected by fluorescence quantitative PCR. Results Compared with the blank group,the apoptosis rate of CD34^+ cells and the protein expressions of p-P38 and p-P53 were significantly decreased in the Compound Shenlu Granule groups with medium-and high-dose,and the Bcl-xl protein and mRNA expressions were significantly up-regulated,with statistically significant differences( P〈0.05). Conclusion The medicated serum of Compound Shenlu Granule can effectively inhibit the TNF-α induced CD34^+ cell apoptosis in normal bone marrow,down-regulate the protein expressions of p-P38 and p-P53 and up-regulate the expression of Bcl-xl. Its mechanism may be related to the P38 MAPKmediated apoptosis pathway.
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