党参多糖对小鼠造血干细胞衰老相关蛋白p53 p21 Bax和Bcl-2的影响
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摘要
目的研究党参多糖对辐射所致小鼠造血干细胞中p53、p21、Bax、Bcl-2蛋白表达的影响以及延缓造血干细胞衰老的可能机制。方法将C57BL/6J小鼠使用随机数字表法分成空白组、模型组和党参多糖低、中、高剂量组。模型组和党参多糖各组采用3Gy/8F强度的X射线均匀照射小鼠,建立造血干细胞衰老模型。党参多糖各组在照射期间分别给小鼠灌胃党参多糖100、200、300mg·kg~(-1),空白组、模型组给予等量生理氯化钠溶液。采用免疫磁珠分离造血干细胞,并检测各组细胞周期变化。用β-半乳糖苷酶染色验证小鼠造血干细胞衰老模型是否成功,用免疫印迹法检测造血干细胞p21、p53、Bax和Bcl-2蛋白表达。结果模型组比其他各组造血干细胞G1期阻滞明显增加(P<0.05);模型组β-半乳糖苷酶染色阳性细胞率明显高于正常组(P<0.05),p53、p21、Bax蛋白表达上调(P<0.05),Bcl-2蛋白表达下调(P<0.05)。党参多糖各组比模型组造血干细胞G1期阻滞明显降低(P<0.05),β-半乳糖苷酶染色阳性率明显低于模型组(P<0.05),p53、p21、Bax蛋白表达下调(P<0.05)、Bcl-2蛋白表达上调(P<0.05)。结论党参多糖能够延缓X线诱导的造血干细胞衰老,其作用机制可能与p53-p21信号通路,Bax与Bcl-2凋亡途径有关。
Objective To study the effect of Codonopsis polysaccharide(CPPS)on the protein expression of p53,p21,Bax and Bcl-2 in hematopoietic stem cells(HSC)induced by radiation,and to investigate the possible mechanism by which CPPS delays the aging of HSC.Methods The C57BL/6Jmice were randomly divided into blank,model and CPPS low,medium and high groups using the random number table method.The model group and CPPS groups were irradiated with X rays of 3Gy/8F,so as to establish the HSC aging model.CPPS groups were respectively given CPPS 100,200 and 300 mg·kg~(-1) during irradiation.The blank group and model group were given an equal volume of normal saline(NS).Immunomagnetic beads method was used for separation and each cell was tested for their cycle changes.Beta-galactosidase(SA-beta-Gal)staining was used to verify the success of HSC aging model in mice.The expression of p21,p53,Bax and Bcl-2 protein in HSC was detected by Western blot.Results Compared with other groups,HSC G1 arrest in the model group was significantly increased(P<0.05).The positive cell rate of SA-beta-Gal in model group was significantly higher than that in normal group(P<0.05),the protein expression of p53,p21,Bax was up-regulated(P<0.05)and the expression of Bcl-2protein was downregulated(P<0.05).HSC G1arrest in CPPS groups was significantly lower(P<0.05)than that in the model group.The positive cell rates of SA-beta-Gal in CPPS group was significantly lower than that of model group(P<0.05),the expression of p53,p21and Bax was down-regulated(P<0.05),and the expression of Bcl-2protein was up-regulated(P<0.05).Conclusion CPPS can delay the aging of HSC induced by X-ray.The mechanism may be related to the signaling pathway of p53-p21,the apoptosis pathway of Bax and Bcl-2.
Objective To study the effect of Codonopsis polysaccharide(CPPS)on the protein expression of p53,p21,Bax and Bcl-2 in hematopoietic stem cells(HSC)induced by radiation,and to investigate the possible mechanism by which CPPS delays the aging of HSC.Methods The C57BL/6Jmice were randomly divided into blank,model and CPPS low,medium and high groups using the random number table method.The model group and CPPS groups were irradiated with X rays of 3Gy/8F,so as to establish the HSC aging model.CPPS groups were respectively given CPPS 100,200 and 300 mg·kg~(-1) during irradiation.The blank group and model group were given an equal volume of normal saline(NS).Immunomagnetic beads method was used for separation and each cell was tested for their cycle changes.Beta-galactosidase(SA-beta-Gal)staining was used to verify the success of HSC aging model in mice.The expression of p21,p53,Bax and Bcl-2 protein in HSC was detected by Western blot.Results Compared with other groups,HSC G1 arrest in the model group was significantly increased(P<0.05).The positive cell rate of SA-beta-Gal in model group was significantly higher than that in normal group(P<0.05),the protein expression of p53,p21,Bax was up-regulated(P<0.05)and the expression of Bcl-2protein was downregulated(P<0.05).HSC G1arrest in CPPS groups was significantly lower(P<0.05)than that in the model group.The positive cell rates of SA-beta-Gal in CPPS group was significantly lower than that of model group(P<0.05),the expression of p53,p21and Bax was down-regulated(P<0.05),and the expression of Bcl-2protein was up-regulated(P<0.05).Conclusion CPPS can delay the aging of HSC induced by X-ray.The mechanism may be related to the signaling pathway of p53-p21,the apoptosis pathway of Bax and Bcl-2.
引文
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[3]Huh CJ,Zhang B,Victor M,et al.Maintenance of age in human neurons generated by microRNA-based neuronal conversion of fibroblasts[J].Elife,2016,5:e18648.DOI:10.7554/eLife.18648
[4]Zou L,Chai J,Gao Y,et al.Down-regulated PLAC8promotes hepatocellular carcinoma cell proliferation by enhancing PI3K/Akt/GSK3β/Wnt/β-catenin signaling[J].Biomed Pharmacother,2016,16(84):139-146
[5]Ramírez-Barrantes R,Marchant I,Olivero P.TRPV1 may increase the effectiveness of estrogen therapy on neuroprotection and neuroregeneration[J].Neural Regen Res,2016,11(8):1204-1207
[6]Bernitz J,Macarthur B,Sieburg H,et al.Hematopoietic stem cells count and remember self-renewal divisions[J].Cell,2016,44(9):S58-S59
[7]陈文霞.纹党多糖硒化物的制备、结构表征及其抗肿瘤活性研究[D].兰州大学,2015
[8]贾道勇,刘俊,李成鹏,等.当归多糖调控人白血病干细胞衰老的机制研究[J].中国中药杂志,2015,40(1):112-117
[9]Wang Y,Schulte BA,LaRue AC,et al.Total body irradiation selectively induces murine hematopoietic stem cell senescence[J].Blood,2006,107(1):358-366
[10]Moehrle BM,Geiger H.Aging of hematopoietic stem cells:DNA damage and mutations[J].Exp Hematol,2016,44(10):895-901
[11]Wang Y,Liu L,Zhou D,et al.Inhibitionofp38MAPK attenu-ates ionizing radiation-induced hematopoietic cell senescence andresidual bone marrow injury[J].Radiat Res,2011,176(6):743-752
[12]Wang Y,Boerma M,Zhou D.Ionizing radiation-induced endothelial cell senescence and cardiovascular diseases[J].Radiat Res,2016,186(2):153-161
[13]张铁辉.IL-6在垂体瘤细胞衰老中表达调控和作用机制研究[D].苏州大学,2014
[14]Sun Y,Jin L,Liu JH,et al.Interfering EZH2expression reverses the cisplatin resistance in human ovarian cancer by inhibiting autophagy[J].Cancer Biother Radiopharm,2016,31(7):246-252
[15]Chen J,Xu T,Zhu D,et al.Egg antigen p40of schistosoma japonicum promotes senescence in activated hepatic stellate cells by activation of the STAT 3/p53/p21pathway[J].Cell Death Dis,2016,6(7):e2315DOI:10.1038/cddis.2016.228
[16]Li J,Cai D,Yao X,et al.Protective effect of ginsenoside Rg1on hematopoietic stem/progenitor cells through attenuating oxidative stress and the Wnt/β-Catenin signaling pathway in a mouse model of d-galactose-induced aging[J].Int J Mol Sci,2016,17(6):849-866
[17]Lu Y,Wang JC,Dapeng C,et al.Bioinformatics analysis of proteomics profiles in senescent human primary proximal tubule epithelial cells[J].BMC Nephrology,2016,17(1):1-8
[18]Chen Y,Pan K,Wang P,et al.HBP1-mediated regulation of p21protein through the Mdm2/p53and TCF4/EZH2pathways and its impact on cell senescence and tumorigenesis[J].JBiol Chem,2016,291(24):12688-12705
[19]Jin H,Lian N,Zhang F,et al.Activation of PPARγ/P53signaling is required for curcumin to induce hepatic stellate cell senescence[J].Cell Death Dis,2016,7(4):e2189
[20]Yang H,Qiu L,Zhang L,et al.Platinum-zoledronate complex blocks gastric cancer cell proliferation by inducing cell cycle arrest and apoptosis[J].Tumour Biol,2016,37(8):10981-10992
[21]Bishayee K,Paul A,Ghosh S,et al.Condurango-glycoside-Afraction of gGonolobus condurango induces DNA damage associated senescence and apoptosis via ROS-dependent p53signalling pathway in HeLa cells[J].Mol Cell Biochem,2013,382(1-2):173-183
[22]Ji J,Tian Y,Zhu Y,et al.Ionizing irradiation inhibits keloid fibroblast cell proliferation and induces premature cellular senescence[J].TJ Dermatol,2015,42(1):56-63
[23]Abramova MVI,Pojidaeva AK,Filippova EA,et al.HDACinhibitors induce apoptosis but not cellular senescence in Gadd45α-deficient E1A+Ras cells[J].Int J Biochem Cell Biol,2014,5(1):102-110