原花青素B1对LPS诱导小鼠巨噬细胞RAW264.7损伤的保护作用及其机制

doi:10.13481/j.1671-587x.20190609

吉林大学学报(医学版) ›› 2019, Vol. 45 ›› Issue (06): 1243-1247.doi: 10.13481/j.1671-587x.20190609

原花青素B1对LPS诱导小鼠巨噬细胞RAW264.7损伤的保护作用及其机制

张宸豪¹, 李瑶¹, 李正祎¹, 骆晓峰²

1. 吉林医药学院检验学院病原生物学教研室, 吉林吉林 132013;
2. 吉林医药学院基础医学院生理学教研室, 吉林吉林 132013

收稿日期:2019-08-24 出版日期:2019-12-05 发布日期:2019-12-05
通讯作者: 骆晓峰,副教授(Tel:0432-64560323,E-mail:zhangchenhao0618@163.com) E-mail:zhangchenhao0618@163.com
作者简介:张宸豪(1972-),男,吉林省吉林市人,讲师,医学博士,主要从事感染免疫相关疾病方面的研究。
基金资助:
国家自然科学基金资助课题（82102953）；吉林省科技厅科研项目资助课题（20170311058YY）；吉林省卫生厅科研项目资助课题（2013Z091）

Protective effect of procyanidine B1 on LPS-induced injury of mouse macrophages RAW264.7 and its mechanism

ZHANG Chenhao¹, LI Yao¹, LI Zhengyi¹, LUO Xiaofeng²

1. Department of Pathogen Biology, College of Medical Laboratory, Jilin Medical College, Jilin 132013, China;
2. Department of Physiology, College of Basic Medical Sciences, Jilin Medical University, Jilin 132013, China

Received:2019-08-24 Online:2019-12-05 Published:2019-12-05

摘要/Abstract

摘要： 目的：观察原花青素B1（PB1）对脂多糖（LPS）诱导的小鼠巨噬细胞RAW264.7损伤的保护作用，探讨其可能的作用机制。方法：体外培养的处于对数生长期的小鼠巨噬细胞RAW264.7分为对照组（细胞不进行处理）、LPS组（细胞给予2 mg·L^-1LPS）、PB1组（细胞给予10 μmol·L^-1 PB1）和PB1+LPS组（细胞给予2 mg·L^-1LPS+10 μmol·L^-1 PB1）。显微镜下观察各组细胞形态表现，流式细胞术检测各组细胞中活性氧（ROS）水平、细胞凋亡率和细胞表面膜分子CD16/32、CD40、CD86及Toll样受体4（TLR4）表达水平。结果：与对照组比较，LPS组细胞皱缩变圆，但PB1组和PB1+LPS组细胞形态表现变化不明显。与对照组比较，LPS组细胞中ROS水平明显升高（P<0.05）；与LPS组比较，PB1+LPS组细胞中ROS水平降低（P<0.05）。与对照组比较，LPS组细胞凋亡率升高（P<0.05）；与LPS组比较，PB1+LPS组细胞凋亡率降低（P<0.05）。与对照组比较，LPS组细胞表面膜分子CD16/32、CD40、CD86和TLR4表达水平升高（P<0.05）；与LPS组比较，PB1+LPS组细胞表面膜分子CD16/32、CD40、CD86和TLR4表达水平降低（P<0.05）。结论：LPS通过诱导细胞中ROS水平升高引起细胞损伤，PB1通过降低ROS水平，下调细胞表面膜分子CD16/32、CD40、CD86和TLR4表达对细胞发挥保护作用。

关键词: 原花青素B1, 巨噬细胞, 脂多糖, 损伤, 细胞凋亡

Abstract: Objective: To observe the protective effect of procyanidin B1 (PB1) on the lipopolysaccharide (LPS)-induced injury of macrophages RAW264.7, and to explore its possible mechanism. Methods: The macrophages RAW264.7 in logarithmic phase cultured in vitro were divided into control group (without treatment), LPS group (treated with 2 mg·L^-1 LPS), PB1 group (treated with 10 μmol·L^-1 PB1) and PB1 + LPS group (treated with 2 mg·L^-1 LPS and 10 μmol·L^-1 PB1). The morphology of cells was observed under microscope, the levels of reactive oxygen species (ROS) in the cells,the apoptotic rates of the cells and the expression levels of CD16/32, CD40, CD86, and Toll-like receptor 4(TLR4) on cell surface of the cells in various groups were detected by flow cytometry. Results: Compared with control group, the cells in LPS group were shrunk and round,but the morphological changes of the cells in PB1 group and PB1 + LPS group were not significant. Compared with control group, the ROS level in the cells in LPS group was significantly increased (P<0.05); compared with LPS group, the ROS level in the cells in PB1 + LPS group was significantly decreased (P<0.05). Compared with control group, the apoptotic rate of the cells in LPS group was decreased (P<0.05); compared with LPS group, the apoptotic rate of the cells in PB1 + LPS group was increased(P<0.05). Compared with control group, the expression levels of cell surface molecules CD16/32, CD40, CD86, and TLR4 in the cells in LPS group were increased (P<0.05); compared with LPS group, the expression levels of cell surface molecules CD16/32, CD40, CD86, and TLR4 in the cells in PB1 + LPS group were decreased (P<0.05). Conclusion: LPS can induce the cell injury by increasing the ROS level, and PB1 can protect the cells by decreasing the ROS level and down-regulating the expression levels of cell surface molecules CD16/32, CD40, CD86, and TLR4.

Key words: procyanidin B1, macrophages, lipopolysaccharide, damage, apoptosis

中图分类号:

R392.28

张宸豪, 李瑶, 李正祎, 骆晓峰. 原花青素B1对LPS诱导小鼠巨噬细胞RAW264.7损伤的保护作用及其机制[J]. 吉林大学学报(医学版), 2019, 45(06): 1243-1247.

ZHANG Chenhao, LI Yao, LI Zhengyi, LUO Xiaofeng. Protective effect of procyanidine B1 on LPS-induced injury of mouse macrophages RAW264.7 and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2019, 45(06): 1243-1247.

参考文献

[1] 周坦洋,罗芙蓉,白彬.葡萄籽原花青素生物药理活性的研究进展[J].哈尔滨医科大学学报,2012,46(1):94-96.
[2] 杨霞,王利,李少伟,等.花青素抗炎机制的研究进展[J].山东医药,2017,57(18),106-108.
[3] 陈菲,盛柳青,麻佳蕾.松针中原花青素的闪式提取及其抗氧化活性[J].中国医药工业杂志,2014,45(2):120-123.
[4] 周静,王轶楠,柳忠辉,等.脂多糖诱导小鼠巨噬细胞系RAW264.7细胞的活化凋亡作用[J].中国生物制品学杂志, 2009,22(2):136-138.
[5] 王艳红,葛斌,杨社华,等.葡萄籽原花青素对复发性结肠炎大鼠结肠组织中细胞因子的影响[J].中国医院药学杂志,2015,35(16):1453-1456.
[6] 李文广,张小郁,吴勇杰,等.葡萄籽中原花青素的抗炎作用和机制[J].中国药理学报,2001,22(12):1117-1120.
[7] 陈晓玲. 植物药中原花青素的抗氧化作用[J].中国中药杂志,2003,28(8):700-702.
[8] MACKENZIE G G,DELFINO J M,KEEN C L,et al.Dimeric procyanidins are inhibitors of NF-kappaB-DNA binding[J].Biochem Pharmacol,2009,78(9):1252-1262.
[9] CHACÓN M R,CEPERUELO-MALLAFRÉ V,MAYMÓ-MASIP E,et al. Grape-seed procyanidins modulate inflammation on human differentiated adipocytes in vitro[J]. Cytokine,2009,47(2): 137-142.
[10] WU H C, JIANG K F, YIN N N,et al.Thymol mitigates lipopolysaccharide-induced endometritis by regulating the TLR4-and ROS-mediated NF-κB signaling pathways[J].Oncotarget,2017,8(12): 20042-20055.
[11] MANDAL A, BHATIA D, BISHAYEE A.Anti-inflammatory mechanism involved in pomegranate-mediated prevention of breast cancer: the role of NF-κB and Nrf2 signaling pathways[J].Nutrients, 2017,9(5): 436.
[12] GASPARRINI M, FORBES-HERNANDEZ T Y, GIAMPIERI F, et al. Anti-inflammatory effect of strawberry extract against LPS-induced stress in RAW 264.7 macrophages[J].Food Chem Toxicol,2017, 102: 1-10.
[13] 孙静文,张伟.复方苦参注射液对巨噬细胞内NF-kB作用的研究[J].现代检验医学杂志,2005(5):24-26.
[14] MA Y F, TANG T, SHENG L L, et al. Aloin suppresses lipopolysaccharide-induced inflammation by inhibiting JAK1-STAT1/3 activation and ROS production in RAW264.7 cells[J].Int J Mol Med,2018, 42(4): 1925-1934.
[15] WEI Z, WANG J, SHI M, et al.Saikosaponin a inhibits LPS-induced inflammatory response by inducing liver X receptor alpha activation in primary mouse macrophages[J].Oncotarget, 2016,7:48995-49007.
[16] 周宪宾,姚成芳.巨噬细胞M1/M2极化分型的研究进展[J].中国免疫学杂志, 2012,28(10):957-960.
[17] 陈方圆,袁祖贻,周娟,等.姜黄素促进RAW264.7源性M1巨噬细胞向替代或激活M2表型极化[J].西安交通大学学报:医学版, 2015,36(2):257-262.
[18] PAN Y, SHEN B, GAO Q,et al. Caspase-1 inhibition attenuates activation of BV2 microglia induced by LPS-treated RAW264.7 macrophages[J].Published Online,2017,26(6): 1633-1640.
[19] GASPARRINI M, FORBES-HERNANDEZ T Y, GIAMPIERI F, et al. Anti-inflammatory effect of strawberry extract against LPS-induced stress in RAW 264.7 macrophages[J].Food Chem Toxicol,2017, 102: 1-10.
[20] KARWASRA R, KALRA P, GUPTA Y K, et al.Antioxidant and anti-inflammatory potential of pomegranate rind extract to ameliorate cisplatin-induced acute kidney injury[J]. Food Funct,2016,7(7): 3091-3101.
[21] XIANG JL, APEA-BAH F B, NDOLO V U, et al.Profile of phenolic compounds and antioxidant activity of finger millet varieties[J].Food Chem,2019,275: 361-368.
[22] 帅维正,刘剑飞,吴成林,等.创伤弧菌溶细胞素对小鼠骨髓来源巨噬细胞坏死性凋亡的作用及机制[J].解放军医学杂志,2018,43(5):419-423.
[23] 刘若鸿,刘春雨,张誉凡,等.早期腹腔穿刺引流对重症急性胰腺炎大鼠腹腔巨噬细胞表型极化的影响[J].解放军医学杂志,2018,43(10):834-839.

原花青素B1对LPS诱导小鼠巨噬细胞RAW264.7损伤的保护作用及其机制

Protective effect of procyanidine B1 on LPS-induced injury of mouse macrophages RAW264.7 and its mechanism

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