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Guilu Erxian Glue (龟鹿二仙胶) Inhibits Chemotherapy-Induced Bone Marrow Hematopoietic Stem Cell Senescence in Mice May via p16INK4a-Rb Signaling Pathway

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Abstract

Objective

To evaluate the effect of Guilu Erxian Glue (龟鹿二仙胶, GEG) on cyclophosphamide (CTX)-induced bone marrow hematopoietic stem cells (HSCs) senescence in mice and explore the underlying mechanism.

Methods

The H22 liver cancer ascites lump model was established in male Kunming mice by injecting intraperitoneally (i.p.) with 5 × 106/mL H22 cells per mouse. Fifty tumor-bearing mice were divided into the control, model, pifithrin-α, GEG, and GEG+pifithrin-α groups using a random number table, 10 mice in each group. CTX (100 mg/kg i.p.) was administrated to mice from day 1 to day 3 (d1–d3) continuously except for the control group. The mice in the pifithrin-α, GEG and GEG+pifithrin-α groups were treated with pifithrin-α (2.2 mg/(kg·d) i.p.) for 6 consecutive days (d4–d9), GEG (9.5 g/(kg·d) i.p.) for 9 consecutive days (d1–d9), and GEG plus pifithrin-α, respectively. HSCs were collected after 9-d drug treatment. The anti-aging effect of GEG was studied by cell viability, cell cycle, and β -galactosidase (β -gal) assays. The mRNA and protein expressions of cyclin-dependent kinase 2 (CDK2), CDK4, inhibitor of cyclin-dependent kinase 4a encoding the tumor suppressor protein p16 (p16INK4a), p21Cip1/Waf1, p53, and phosphorylated retinoblastoma (pRb) were evaluated by quantitative real-time reverse transcription-polymerase chain reaction and semi-quantitative Western blot, respectively.

Results

Compared with the model group, GEG increased cell viability as well as proliferation (P<0.05 or P<0.01) and reduced β -gal expression. Furthermore, GEG significantly decreased the expressions of p16INK4a, p53 and p21Cip1/Waf1 proteins, and increased the expressions of CDK2, CDK4 and pRb proteins compared with the model group (P<0.05 or P<0.01).

Conclusion

GEG can alleviate CTX-induced HSCs senescence in mice, and the p16INK4a-Rb signaling pathway might be the underlying mechanism.

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References

  1. Karol SE, Yang W, Smith C, Cheng C, Stewart C, Baker SD, et al. Palmar-plantar erythrodysesthesia syndrome following treatment with high-dose methotrexate or highdose cytarabine. Cancer 2017;123:3602–3608.

    Article  CAS  Google Scholar 

  2. Kumar KB, Kuttan R. Chemoprotective activity of an extract of Phyllanthus amarus against cyclophosphamide induced toxicity in mice. Phytomedicine 2005;12:494–500.

    Article  CAS  Google Scholar 

  3. Chang JY, Brown S. Cytosine arabinoside differentially alters survival and neurite outgrowth of neuronal PC12 cells. Biochem Biophys Res Commun 1996;218:753–758.

    Article  CAS  Google Scholar 

  4. Basu A, Ghosh P, Bhattacharjee A, Patra AR, Bhattacharya S. Prevention of myelosuppression and genotoxicity induced by cisplatin in murine bone marrow cells: effect of an organovanadium compound vanadium (III)-L cysteine. Mutagenesis 2015;30:509–517.

    Article  CAS  Google Scholar 

  5. Henricks LM, Opdam FL, Beijnen JH, Cats A, Schellens JHM. DPYD genotype-guided dose individualization to improve patient safety of fluoropyrimidine therapy: call for a drug label update. Ann Oncol 2015;82:153–160.

    Google Scholar 

  6. Othieno-Abinya NA, Waweru A, Nyabola LO. Chemotherapy induced myelosuppression. East Afr Med J 2007;84:8–15.

    Article  CAS  Google Scholar 

  7. Lu L, Wang YY, Zhang JL, Li DG, Meng AM. p38 MAPK inhibitor insufficiently attenuates HSC senescence administered long-term after 6 Gy total body irradiation in mice. Int J Mol Sci 2016;17:905–914.

    Article  Google Scholar 

  8. Toussaint O, Medrano EE, von Zglinicki T. Cellular and molecular mechanisms of stress-induced premature senescence (SIPS) of human diploid fibroblasts and melanocytes. Exp Gerontol 2000;35:927–945.

    Article  CAS  Google Scholar 

  9. Pant V, Quintás-Cardama A, Lozano G. The p53 pathway in hematopoiesis: lessons from mouse models, implications for humans. Blood 2012;120:5118–5127.

    Article  CAS  Google Scholar 

  10. Perez-Campo FM, Costa G, Lie-A-Ling M, Stifani S, Kouskoff V, Lacaud G. MOZ-mediated repression of p16INK4a is critical for the self-renewal of neural and hematopoietic stem cells. Stem Cells 2014;32:1591–1601.

    Article  CAS  Google Scholar 

  11. Yue Z, Rong J, Ping W, Bing Y, Xin Y, Feng LD, et al. Gene expression of the p16INK4a-Rb and p19Arf-p53-p21Cip/Waf1 signaling pathways in the regulation of hematopoietic stem cell aging by ginsenoside Rg1. Genet Mol Res 2014;13:10086–10096.

    Article  CAS  Google Scholar 

  12. Wang LF, Xu ZY, Wang ZQ, Zhang M, Yan GY, Hou AJ, et al. Clinical observation of Shuanghuang Shengbai Granule on prevention and treatment of myelosuppression caused by chemotherapy in cancer patients. Chin J Integr Med 2017;23:105–109.

    Article  Google Scholar 

  13. Sun X, Zhao YN, Qian S, Gao RL, Yin LM, Wang LP, et al. Ginseng-derived panaxadiol saponins promote hematopoiesis recovery in cyclophosphamide-induced myelosuppressive mice: potential novel treatment of chemotherapy-induced cytopenias. Chin J Integr Med 2018;24:200–206.

    Article  CAS  Google Scholar 

  14. Wang J, Wei DN, Zhang WP, Ran R, Xu K, Gao JW, et al. Adjuvant function of Guilu Erxian Glue cataplasm in treating carcinoma of the large intestine patients with myelosuppression after chemotherapy: a clinical observation. Chin J Integr Tradit West Med (Chin) 2014;34:947–951.

    Google Scholar 

  15. Wang B, Zhang MZ, Fu XR, Liu XY, Zhang L. Research progress on the mechanism and prevention of bone marrow suppression by chemoradiation. J Basic Clin Oncol 2013;26:162–165.

    Google Scholar 

  16. Wang J, Lu X, Sakk V, Klein CA, Rudolph KL. Senescence and apoptosis block hematopoietic activation of quiescent hematopoietic stem cells with short telomeres. Blood 2014;124:3237–3240.

    Article  CAS  Google Scholar 

  17. Janzen V, Forkert R, Fleming HE, Saito Y, Waring MT, Dombkowski DM, et al. Stem-cell ageing modified by the cyclin-dependent kinase inhibitor p16INK4a. Nature 2006;443:421–426.

    Article  CAS  Google Scholar 

  18. Lin SY, Shen MH, Liu ZD. Study on resistance effects of Guilu Erxian Jiao on apoptosis of stem CD34+cells in mice with chemotherapy. Chin J Tradit Med Sci Technol (Chin) 2008;15:172–173.

    Google Scholar 

  19. Roemer K. Notch and the p53 clan of transcription factors. Adv Exp Med Biol 2012;727:223–240.

    Article  CAS  Google Scholar 

  20. Zubbair Malik M, Ali S, Alam MJ, Ishrat R, Brojen Singh RK. Dynamics of p53 and Wnt cross talk. Comput Biol Chem 2015;59Pt B:55–66.

    Article  CAS  Google Scholar 

  21. Liu H, Jia D, Li A, Chau J, He D, Ruan X, et al. p53 regulates neural stem cell proliferation and differentiation via BMP-Smad1 signaling and Id1. Stem Cells Dev 2013;22:913–927.

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Oncology, the Third Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, 310005, China

    Jue Wang, Yin-yin Ying & Wei-ping Zhang

  2. Department of Oncology, Ningbo Forth Hospital, Ningbo, Zhejiang Province, 315700, China

    Zhao-hui Chen

  3. Science Research Department, Zhejiang Chinese Medical University, Hangzhou, 310053, China

    Ke-ding Shao

  4. Department of Oncology, Hangzhou Hospital of Traditional Chinese Medicine, Hangzhou, 310007, China

    Sheng-you Lin

Authors
  1. Jue Wang
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  2. Yin-yin Ying
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  3. Zhao-hui Chen
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  4. Ke-ding Shao
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  5. Wei-ping Zhang
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  6. Sheng-you Lin
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Contributions

Wang J contributed to execution of study, acquisition of data, and manuscript drafting. Ying YY and Chen ZH performed the experiments and statistical analysis; Shao KD and Zhang WP contributed to data analysis and critical discussion. Lin SY contributed to presenting conception and design of the study, drafting and revising the manuscript, and approving final version for publication.

Corresponding author

Correspondence to Sheng-you Lin.

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Conflict of Interest

All authors have no conflicts of interest to declare.

Supported by the National Natural Science Foundation of China (No. 81904197), Natural Science Foundation of Zhejiang Province (No. LQ15H290002), and 2019 Research and Innovation Fund Project for Young and Middle-aged Researchers of Zhejiang Chinese Medical University (No. KC201944)

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Wang, J., Ying, Yy., Chen, Zh. et al. Guilu Erxian Glue (龟鹿二仙胶) Inhibits Chemotherapy-Induced Bone Marrow Hematopoietic Stem Cell Senescence in Mice May via p16INK4a-Rb Signaling Pathway. Chin. J. Integr. Med. 26, 819–824 (2020). https://doi.org/10.1007/s11655-020-3098-3

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  • DOI: https://doi.org/10.1007/s11655-020-3098-3

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