人参皂苷Rg1对衰老模型大鼠骨髓造血功能的影响及其机理

doi:10.13523/j.cb.20150803

中国生物工程杂志

2015, Vol. 35

Issue (8): 16-22 DOI: 10.13523/j.cb.20150803

研究报告

人参皂苷Rg1对衰老模型大鼠骨髓造血功能的影响及其机理

邵月, 张力恒, 冉瑞图, 孙嘉政, 张晶, 张梦思, 贾道勇, 张岩岩, 王亚平

重庆医科大学干细胞与组织工程研究室重庆 400016

Effect of Ginsenoside Rg1 on Bone Marrow Hematopietic Function of Aging Model Rats and Its Biological Mechanism

SHAO Yue, ZHANG Li-heng, RAN Rui-tu, SUN Jia-zheng, ZHANG Jing, ZHANG Meng-si, JIA Dao-yong, ZHANG Yan-yan, WANG Ya-ping

Laboratory of Stem Cell and Tissue Engineering, Chongqing Medical University, Chongqing 400016, China

全文: PDF(856 KB) HTML

摘要：

目的: 探讨人参皂苷Rg1对衰老模型大鼠骨髓造血功能的影响及其机理。方法: SD大鼠随机分为4组,每组10只。衰老模型组,皮下注射D-半乳糖120mg/kg qd×42;Rg1衰老模型组,注射D-半乳糖剂量与时间同衰老模型组,第15d起腹腔注射Rg1 20mg/kg qd×28;正常对照组,皮下注射生理盐水qd×42;Rg1正常对照组,注射生理盐水qd×14,第15d起腹腔注射Rg1(同Rg1衰老模型组)。模型复制或药物注射完成后第2d,采外周血检测血白细胞总数与分类计数和晚期糖基化终产物(AGEs),骨髓单个核细胞(BMNCs)计数,衰老相关β-半乳糖苷酶(SA-β-Gal)染色检测BMNCs衰老百分率,多向造血祖细胞集落(CFU-Mix)培养检测BMMCs形成集落能力;Western blotting 检测BMMCs衰老相关蛋白P21和P53的变化;提取腹腔巨噬细胞进行培养,比色法检测巨噬细胞吞噬功能。结果:与衰老模型组比较, Rg1衰老模型组外周血白细胞数量增多,淋巴细胞比例升高,粒细胞比例降低,CD8⁺T细胞所占比例降低,CD4⁺T细胞所占比例升高,外周血AGEs水平降低,每根股骨的BMMCs细胞数增多, SA-β-Gal染色阳性的BMMCs明显降低,BMMCs形成CFU-Mix能力提高,P21和P53的表达下调,腹腔巨噬细胞吞噬中性红指数上升。结论: D-半乳糖复制的衰老模型大鼠骨髓造血功能损伤明显,人参皂苷Rg1对其致衰损伤有明确的保护作用,可能机理与调控p53/p21信号通路有关。

关键词： 人参皂苷Rg1; 衰老模型; 骨髓; 造血细胞; 机理

Abstract:

Objective: To explore the effects and the relative mechanism of ginsenoside Rg1 on bone marrow hematopietic function in aging model rats, and provide theoretical and experimental evidences for searching effective natural medicine of antiaging or protecting hematopietic function. Methods: Male SD rats were randomly divided into normal group, Rg1 normal group, aging model group and Rg1 aging model group. Aging model group: the rats were administrated with D-galactose 120mg/kg,qd×42 by subcutaneous way; Rg1 aging group: the rats were also given D-galactose of the same dose and time as aging model group, and from the 15th day on, rats were treated with Rg1 20 mg/kg qd×28 by intraperitoneal way; Normal control group: The rats were injected with same volume saline qd×42 by subcutaneous way; Rg1 normal group: the rats were given saline with the same volume,qd×14, then received Rg1 20 mg/kg qd×28 by same way. After 2 days of finishing the treatment, peripheral blood was collected to detect the number of leukocytes,analyzes classification of leukocyte and advanced glycosylation end products (AGEs); The femur was taken to count the number of bone marrow mononuclear cells(BMMCs) in each femur, the proliferation of BMMCs was detected; Senescence-accociated β-galactosidase (SA-β-Gal) stain was analyzed aging BMMCs; multipotential hematopoietic progenitor (CFU-Mix) was cultured to observe the colony formation of BMMCs; the expression of senescence-associated protein P21 and P53 in BMMCs was assayed by Western blotting; peritoneal macrophage was extracted and detected the phagocytosis of the macrophage by colorimetric method. Results: Compared with the aging model group, Rg1 aging model group can significantly increase the number of leukocytes, regulate the percentage of granulocyte and lymphocytes, the proportion of CD8⁺T cells and CD4⁺T cells; the number of BMMCs in each femur; decrease the ratio of SA-β-Gal positive BMMCs,the amounts of AGEs;enhance the ability of BMMCs to form CFU–Mix; down-regulate the expression of senescence- associated protein P21 and P53;promote the phagocytic index of peritoneal macrophage. Conclusion: The hematopoietic cells are obviously induced senescence by treating with D-galactose, ginsenoside Rg1 has a significantly antiaging or protective effect on hematopoietic cells. It is suggested that the mechanism may be Rg1 protecting hematopietic cells by regulating p53/p21 signaling pathway.

Key words: Ginsenoside Rg1 Aging model Bone marrow Hematopoietic cell Mechanism

收稿日期: 2015-03-28 出版日期: 2015-08-25

ZTFLH:

R329.2

基金资助:

国家自然科学基金(30973818),国家教育部博士导师基金(20125503110006)资助项目

通讯作者: 王亚平 E-mail: ypwangcq@aliyun.com

	服务
	把本文推荐给朋友
	加入引用管理器
	E-mail Alert
	RSS
	作者相关文章

引用本文:

邵月, 张力恒, 冉瑞图, 孙嘉政, 张晶, 张梦思, 贾道勇, 张岩岩, 王亚平. 人参皂苷Rg1对衰老模型大鼠骨髓造血功能的影响及其机理[J]. 中国生物工程杂志, 2015, 35(8): 16-22.

SHAO Yue, ZHANG Li-heng, RAN Rui-tu, SUN Jia-zheng, ZHANG Jing, ZHANG Meng-si, JIA Dao-yong, ZHANG Yan-yan, WANG Ya-ping . Effect of Ginsenoside Rg1 on Bone Marrow Hematopietic Function of Aging Model Rats and Its Biological Mechanism. China Biotechnology, 2015, 35(8): 16-22.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20150803 或 https://manu60.magtech.com.cn/biotech/CN/Y2015/V35/I8/16

[1] Wagner W M, Ouyang Q, Sekeri-Pataryas K, et al. Basic biology and clinical impact of immunosenescence. Biogerontology, 2004, 5(1): 63-66.

[2] Chu S, Zhang J.New achievements in ginseng research and its future prospects.Chinese Journal of Integrative Medicine, 2009, 15(6):403-408.

[3] 周玥, 杨斌, 姚欣, 等. 人参皂苷 Rg1 延缓造血干细胞衰老与 p16INK4a 表达关系的研究.中国中药杂志,2011,36(5):608-613. Zhou Y,Yang B,Yao X,et al. Experimental study of relationship between effect of ginsenoside Rg1 to delay hematopoietic stem cell senescence and expression of p16 INK4a.China Journal of Chinese Materia Medica, 2011, 36(5):608-613.

[4] Shang Y Z,Gong M Y,Zhou X X, et al.Improving effects of SSF on memory deficits and pathological changes of neural and immunological systems in senescent mice.Acta Pharmacol Sin, 2001,22(12):1078-1083.

[5] 秦红兵,杨朝晔,范忆江,等.半乳糖诱导衰老小鼠模型的建立与评价.中国组织工程研究与临床康复,2009,13(7):1275-1278. Qin H B, Yang Z Y, Fan Y J,et al. Establishment and evaluation of aging models induced by D-galactose in mice, Journal of Clinical Rehabilitative Tissue Engineering Research, 2009, 13(7):1275-1278.

[6] 彭彬, 陈茂山, 蒲莹, 等.人参皂苷Rg1延缓D-半乳糖大鼠脑衰老作用及机制的初步研究.重庆医科大学学报, 2011, 0(4):419-422. Peng B,Chen M S,Pu Y,et al. Anti- aging effects of ginsenoside Rgl and it's mechanisms on brain aging rats induced by D-galactose. Journal of Chongqing Medical University, 2011, 36(4):419-422.

[7] 郑敏, 王亚平.当归多糖对人髓系多向造血祖细胞增殖分化的影响及其机理研究.解剖学杂志,2002,25(2):105-109. Zheng M,Wang Y P. Study on biological mechanism of angelica polysaccharide regulation on human early multipotentiai progenitor cell. Chinese Journal of Anatomy, 2002,25(2):105-109.

[8] Kobari L,Giarratana M C,Poloni A,et al. Flt3 ligand,MGDF,Epo and GCSF enhance ex vivo expansion of hematopoietic cell compartments in the presence of SCF,IL -3 and IL-6.Bone Marrow Transpl, 1998, 2l(8):759-767.

[9] 孙缅恩, 杜冠华.晚期糖基化终产物的病理意义及其机制.中国药理学通报,2002,18(3):246-249. Sun M E, Du G H. Pathological significance and mechanism of advanced glycation end-products. Chinese Pharmacological Bulletin, 2002, 18(3):246-249.

[10] 杨继乐, 张莉, 王莉.单核-巨噬细胞的分化和功能研究进展.细胞与分子免疫学杂志, 2014,30(11):1213-1216. Yang J L,Zhang L,Wang L. The research progress of mononuclear macrophages' differentiation and function. Journal of Cellular and Molecular Immunology, 2014, 30(11):1213-1216.

[11] Ohtani N, Yamakoshi K, Takahashi A, et al. The p16INK4a-RB pathway: molecular link between cellular senescence and tumor suppression.The Journal of Medical Investigation, 2004, 51(3-4):146-153.

[12] Chen X, Zhang W, Gao Y, et al.Senescence-like changes induced by expression of p21(waf1cip1)in NIH3T3 cell line.Cell Res, 2002, 12(3-4):229-233.

[13] Molchadsky A, Shats I, Goldfinger N, et al. p53 plays a role in mesenchymal differentiation programs, in a cell fate dependent manner.PLoS One, 2008, 3(11):3707-3722.

[1]	赵久梅,王哲,李学英. 调控软骨形成的信号通路及相关因子在骨髓间充质干细胞骨向分化中的作用^*[J]. 中国生物工程杂志, 2021, 41(10): 62-72.
[2]	朱小丽,黄翠,马丽丽,张超,巩玥,赵婉雨,赵秀芳,郭文姣,彭皓,张吉,梁慧刚. 新型冠状病毒病(COVID-19)研究进展[J]. 中国生物工程杂志, 2020, 40(1-2): 38-50.
[3]	袁雅红, 赵珊珊, 王小莉, 腾智平, 李东升, 曾毅. HIV-1 Tat蛋白抑制骨髓间充质干细胞的造血支持功能[J]. 中国生物工程杂志, 2017, 37(6): 1-8.
[4]	赵正德, 陈振银, 张慧楠, 龚剑萍, 许少丹, 罗忠礼. 自组装短肽水凝胶支架三维培养环境对骨髓间充质干细胞生物学特性及心肌方向分化的影响[J]. 中国生物工程杂志, 2017, 37(11): 45-51.
[5]	沈鹏飞, 王斌, 谢子康, 郑冲, 瞿玉兴. 软骨寡聚基质蛋白过表达对BMP-2诱导骨髓间充质干细胞分化的影响[J]. 中国生物工程杂志, 2016, 36(10): 1-7.
[6]	张登央, 张英, 张丽君, 王妍. 3D技术制备骨修复生物材料的功能和安全性评价[J]. 中国生物工程杂志, 2015, 35(7): 55-61.
[7]	黄小娟, 王鸣刚, 廖世奇, 张虹, 王炯蓉, 马瑾, 杨楠, 马宁. 中断BMSCs向神经细胞诱导分化的细胞的恶变研究[J]. 中国生物工程杂志, 2014, 34(3): 26-33.
[8]	刘奔, 张彩顺, 高绪斌, 李桦楠, 孔旭. 聚己内酯/壳聚糖神经导管复合骨髓间充质干细胞促进大鼠坐骨神经损伤修复的研究[J]. 中国生物工程杂志, 2014, 34(2): 34-38.
[9]	陈宇婷, 王长海, 严秀文, 黎军胜. 抗菌肽的设计及其应用[J]. 中国生物工程杂志, 2013, 33(7): 97-102.
[10]	张浩然, 曾志勇, 陈君敏. 体外RNA干扰下调DEPTOR表达对人多发性骨髓瘤细胞增殖和凋亡能力的影响[J]. 中国生物工程杂志, 2013, 33(5): 13-21.
[11]	谢荣辉, 殷嫦嫦, 殷明, 陈伟才, 邬亚华, 何丁文, 林思文, 耿书国. N-乙酰半胱氨酸通过激活PI3K-Akt信号途径抑制过氧化氢诱导骨髓间充质干细胞凋亡[J]. 中国生物工程杂志, 2013, 33(11): 1-7.
[12]	赵德超, 付绍印, 吕晓曼, 吴江鸿, 张燕军, 李金泉, 王志新, 张永斌, 张文广. 褪黑素对绒山羊皮肤FGF5等绒毛生长相关基因的影响[J]. 中国生物工程杂志, 2012, 32(6): 20-26.
[13]	谢佳瑛, 胥文春, 徐道晶, 张晓艳, 唐敏. Notch信号参与BMP9诱导的间充质干细胞成骨分化及其机制的初步探讨[J]. 中国生物工程杂志, 2012, 32(11): 14-22.
[14]	谢佳瑛, 胥文春, 徐道晶, 张晓艳, 唐敏. Notch信号参与BMP9诱导的间充质干细胞成骨分化及其机制的初步探讨[J]. 中国生物工程杂志, 2012, 32(11): 14-22.
[15]	牛红星, 穆军升, 张健群, 胡平, 伯平, 王艳. 骨髓间充质干细胞与聚3-羟基丁酸酯-co-4-羟基丁酸酯生物材料共培养细胞补片的初步研究[J]. 中国生物工程杂志, 2012, 32(10): 74-79.

Viewed

Full text

Abstract

Cited

Shared

Discussed