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On-line Access: 2022-06-08

Received: 2021-12-01

Revision Accepted: 2022-02-16

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Citations:  Bibtex RefMan EndNote GB/T7714

 ORCID:

Yinghua XU

https://orcid.org/0000-0003-2245-3249

Xuemei LU

https://orcid.org/0000-0002-0571-1472

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Journal of Zhejiang University SCIENCE B 2022 Vol.23 No.6 P.481-501

http://doi.org/10.1631/jzus.B2100988


Spirulina platensis aqueous extracts ameliorate colonic mucosal damage and modulate gut microbiota disorder in mice with ulcerative colitis by inhibiting inflammation and oxidative stress


Author(s):  Jian WANG, Liqian SU, Lun ZHANG, Jiali ZENG, Qingru CHEN, Rui DENG, Ziyan WANG, Weidong KUANG, Xiaobao JIN, Shuiqing GUI, Yinghua XU, Xuemei LU

Affiliation(s):  Guangdong Provincial Key Laboratory of Pharmaceutical Bioactive Substances, School of Life Science and Biopharmaceutics, Guangdong Pharmaceutical University, Guangzhou 510006, China; more

Corresponding email(s):   luxuemei@gdpu.edu.cn, xuyh@nifdc.org.cn

Key Words:  Spirulina platensis aqueous extracts, Ulcerative colitis, Oxidative stress, Inflammation, Antioxidant, Gut microbiota


Jian WANG, Liqian SU, Lun ZHANG, Jiali ZENG, Qingru CHEN, Rui DENG, Ziyan WANG, Weidong KUANG, Xiaobao JIN, Shuiqing GUI, Yinghua XU, Xuemei LU. Spirulina platensis aqueous extracts ameliorate colonic mucosal damage and modulate gut microbiota disorder in mice with ulcerative colitis by inhibiting inflammation and oxidative stress[J]. Journal of Zhejiang University Science B, 2022, 23(6): 481-501.

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author="Jian WANG, Liqian SU, Lun ZHANG, Jiali ZENG, Qingru CHEN, Rui DENG, Ziyan WANG, Weidong KUANG, Xiaobao JIN, Shuiqing GUI, Yinghua XU, Xuemei LU",
journal="Journal of Zhejiang University Science B",
volume="23",
number="6",
pages="481-501",
year="2022",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B2100988"
}

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%T Spirulina platensis aqueous extracts ameliorate colonic mucosal damage and modulate gut microbiota disorder in mice with ulcerative colitis by inhibiting inflammation and oxidative stress
%A Jian WANG
%A Liqian SU
%A Lun ZHANG
%A Jiali ZENG
%A Qingru CHEN
%A Rui DENG
%A Ziyan WANG
%A Weidong KUANG
%A Xiaobao JIN
%A Shuiqing GUI
%A Yinghua XU
%A Xuemei LU
%J Journal of Zhejiang University SCIENCE B
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T1 - Spirulina platensis aqueous extracts ameliorate colonic mucosal damage and modulate gut microbiota disorder in mice with ulcerative colitis by inhibiting inflammation and oxidative stress
A1 - Jian WANG
A1 - Liqian SU
A1 - Lun ZHANG
A1 - Jiali ZENG
A1 - Qingru CHEN
A1 - Rui DENG
A1 - Ziyan WANG
A1 - Weidong KUANG
A1 - Xiaobao JIN
A1 - Shuiqing GUI
A1 - Yinghua XU
A1 - Xuemei LU
J0 - Journal of Zhejiang University Science B
VL - 23
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EP - 501
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PB - Zhejiang University Press & Springer
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DOI - 10.1631/jzus.B2100988


Abstract: 
ulcerative colitis (UC) is a chronic and recurrent inflammatory bowel disease (IBD) that has become a major gastroenterologic problem during recent decades. Numerous complicating factors are involved in UC development such as oxidative stress, inflammation, and microbiota disorder. These factors exacerbate damage to the intestinal mucosal barrier. Spirulina platensis is a commercial alga with various biological activity that is widely used as a functional ingredient in food and beverage products. However, there have been few studies on the treatment of UC using S. platensis aqueous extracts (SP), and the underlying mechanism of action of SP against UC has not yet been elucidated. Herein, we aimed to investigate the modulatory effect of SP on microbiota disorders in UC mice and clarify the underlying mechanisms by which SP alleviates damage to the intestinal mucosal barrier. Dextran sulfate sodium (DSS) was used to establish a normal human colonic epithelial cell (NCM460) injury model and UC animal model. The mitochondrial membrane potential assay 3-‍‍(4,5-dimethylthiazol-2-yl)-2,‍5-diphenyltetrazolium bromide (MTT) and staining with Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) and Hoechst 33258 were carried out to determine the effects of SP on the NCM460 cell injury model. Moreover, hematoxylin and eosin (H&E) staining, transmission electron microscopy (TEM), enzyme-linked immunosorbent assay (ELISA), quantitative real-time polymerase chain reaction (qPCR), western blot, and 16S ribosomal DNA (rDNA) sequencing were used to explore the effects and underlying mechanisms of action of SP on UC in C57BL/6 mice. In vitro studies showed that SP alleviated DSS-induced NCM460 cell injury. SP also significantly reduced the excessive generation of intracellular reactive oxygen species (ROS) and prevented mitochondrial membrane potential reduction after DSS challenge. In vivo studies indicated that SP administration could alleviate the severity of DSS-induced colonic mucosal damage compared with the control group. Inhibition of inflammation and oxidative stress was associated with increases in the activity of antioxidant enzymes and the expression of tight junction proteins (TJs) post-SP treatment. SP improved gut microbiota disorder mainly by increasing antioxidant enzyme activity and the expression of TJs in the colon. Our findings demonstrate that the protective effect of SP against UC is based on its inhibition of pro-inflammatory cytokine overproduction, inhibition of DSS-induced ROS production, and enhanced expression of antioxidant enzymes and TJs in the colonic mucosal barrier.

钝顶螺旋藻水提取物通过抑制炎症、氧化应激及调节肠道菌群来改善溃疡性结肠炎小鼠结肠黏膜损伤

王建1*,粟丽千1,2*,张伦1,曾佳利1,陈晴汝1,邓蕊1,王子琰1,邝伟东1,金小宝1,桂水清3
徐颖华4,卢雪梅1
1广东药科大学生命科学与生物制药学院,广东省生物活性药物研究重点实验室,中国广州市,510006
2广东药科大学药学院,中国广州市,510006
3深圳大学第一附属医院,深圳市第二人民医院重症急救医学部,中国深圳市,518031
4中国食品药品检定研究院,卫生部生物技术产品检定方法及其标准化重点实验室,中国北京市,102629
目的:探究钝顶螺旋藻水提取物(SP)对葡聚糖硫酸钠(DSS)诱导的溃疡性结肠炎(UC)小鼠肠道菌群紊乱的调节作用,并阐明其减轻结肠黏膜屏障损伤的潜在机制。
创新点:(1)成功鉴定出钝顶螺旋藻粉末及SP的化学成分;(2)研究结果证实SP可改善UC小鼠的结肠黏膜损伤,并能够调节肠道菌群,SP对UC的保护作用机制是减少炎症因子的过量产生、抑制氧化应激和增强结肠黏膜屏障中紧密连接蛋白(TJ)的表达。(3)SP可作为一种治疗UC、肠道菌群紊乱或氧化应激相关疾病的天然药物,并为该药物临床应用提供理论依据。
方法:采用DSS建立正常肠上皮细胞(NCM460)损伤模型和UC动物模型。采用噻唑兰法(MTT)、细胞凋亡染色法(AnnexinV-FITC/PI)、细胞染色法(Hoechst 33258)和线粒体膜电位法(MMP)测定SP对NCM460细胞损伤模型的影响。并采用苏木精&伊红染色法(H&E)、透射电镜(TEM)、酶联免疫吸附测定法(ELISA)、实时荧光定量聚合酶链式反应(qPCR)、蛋白质印迹法(Western blot)和16S rDNA测序等方法,探讨SP对UC小鼠的结肠黏膜损伤和肠道菌群的影响及其潜在作用机制。
结论:体外研究表明,SP可减轻DSS诱导的NCM460细胞损伤;SP还能显著减少该细胞内活性氧(ROS)的过度产生,并抑制DSS刺激后引起的细胞线粒体膜电位降低。体内研究表明,SP可减轻DSS诱导的结肠黏膜损伤程度。SP治疗后,炎症和氧化应激水平随着抗氧化酶和TJ表达的升高而降低。此外,SP主要通过增加抗氧化酶活性和结肠中TJ的表达来改善肠道菌群紊乱。综上所述,SP对UC的保护作用是基于抑制促炎细胞因子的过量产生、抑制ROS产生、增强结肠黏膜屏障中抗氧化酶和TJ的表达实现。

关键词:钝顶螺旋藻水提取物;溃疡性结肠炎;氧化应激;炎症;抗氧化剂;肠道菌群

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

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