Hu S, et al. J Agric Food Chem. 2020 Sep 2;68(35):9415-9426.
Zhang X, et al. Bioact Mater. 2020 Sep 9;6(2):472-489.
Xin Y, et al. Immunol Lett. 2020 Sep;225:66-73.
Yang Y, et al. Gut. 2020 Oct 29;70(8):1495–506.
Yang Y, et al. Gut. 2020 Oct 29;70(8):1495–506.
Cheng B, et al. Chemosphere. 2021 Jan;263:127849.
Jia M,et al. Talanta. 2020 Aug 15;216:120926.
本品来源于Escherichia coli 055:B5。可用于诱导细胞或者诱导疾病研究的动物模型如炎症反应等相关实验。
Lipopolysaccharides (LPS) 脂多糖是革兰氏阴性细菌细胞壁中的一种特有成分，位于细胞壁的最外层并暴露于非荚膜细菌的细胞表面，有利于维持细胞外膜的完整性，保护细菌免受胆汁盐和脂类抗生素的破坏。结构上，脂多糖由类脂A、核心多糖和O-多糖侧链组成。其中类脂质A是构成细菌内毒素的主要成分，决定其毒性强弱；而O-多糖侧链在不同细菌间是高度变化的，特异性决定细菌的血清型。LPS可以引起免疫刺激的级联反应和机体的毒性病理生理活动.
使用本产品的应用案例（仅供参考）
In Vitro
Cell（RAW264.7 cells，1 μg/mL of LPS，48h）
RAW264.7 cells were incubated in DMEM medium (containing 1 μg/mL of LPS) for 48 hto generate M1 macrophages, and were treated with 50 ng/mL of IL-4 for 24 h to obtain M2 macrophages. Then, the resulted macrophages were stained with CD86/PE (dilution 1:200) and CD206/FITC (dilution 1:100) double dye for 30 min. After being washed twice with PBS, cells were fixed with 4% paraformaldehyde for 10 min and then stained with 10 μg/mL DAPI for 10 min. The staining results were observed via confocal laser scanning microscope.
来源文章：Zhang X, Tang J, Li C, Lu Y, Cheng L, Liu J. A targeting black phosphorus nanoparticle based immune cells nano-regulator for photodynamic/photothermal and photo-immunotherapy. Bioact Mater. 2020 Sep 9;6(2):472-489. doi: 10.1016/j.bioactmat.2020.08.024. PMID: 32995674; PMCID: PMC7493086.
Cell（HepG2 cells，1 μg/mL LPS，12h）
To monitor endogenous ClO−, HepG2 cells were treated with LPS(1 μg/mL) for 12 h and then coincubated with PMA (10 nmol/L) and the peptide@Ag/Au NCs solution at 37 °C for 1 h, followed by washing three times before imaging. In the control assay, HepG2 cells were treated with LPS (1 μg/mL) for 12 h and then incubated with PMA (10 nmol/L) for 1 h. The cells were subsequently cultured in medium containing uric acid (250 nmol/L) and DMSO (0.5%) for 15 min and then treated with the peptide@Ag/Au NCs solution at 37 °C for 1 h. The cells were washed three times with PBS to remove the unbound peptide@Ag/Au NCs and were then observed under a Nikon A1R MP multiphoton microscope with a 60× oil-immersion objective lens. The images of peptide@Ag/Au NCs were captured under excitation at409 nm.
来源文献：Jia M, Mi W, Guo S, Yang QZ, Jin Y, Shao N. Peptide-capped functionalized Ag/Au bimetal nanoclusters with enhanced red fluorescence for lysosome-targeted imaging of hypochlorite in living cells. Talanta. 2020 Aug 15;216:120926. doi: 10.1016/j.talanta.2020.120926. Epub 2020 Mar 14. PMID: 32456892.
Cell（RAW264.7 cells，1 μg/mL of LPS，24h）
For inflammatory experiments, the cells were seeded in a 12-well plate (4 × 105 cells/mL for F4 subfraction and 2 × 105 cells/mL for synthesized peptides, 1 mL/well) and allowed to adhere for 24 h. Afterward, the cells were pretreated for 1 h with the F4 subfraction dissolved in serum-free medium at 25−100 μg/mL. Thereafter, the cells were stimulated with 1 μg/mL LPS for 24 h. Finally, the media were collected, and nitric oxide (NO), tumor necrosis factor (TNF)-α, and interleukin (IL)-6 levels were quantified.
来源文章：Hu S, Yuan J, Gao J, Wu Y, Meng X, Tong P, Chen H. Antioxidant and Anti-Inflammatory Potential of Peptides Derived from In Vitro Gastrointestinal Digestion of Germinated and Heat-Treated Foxtail Millet (Setaria italica) Proteins. J Agric Food Chem. 2020 Sep 2;68(35):9415-9426. doi: 10.1021/acs.jafc.0c03732. Epub 2020 Aug 18. PMID: 32786864.
Cell（RAW264.7 cells，0.5 μg/mL of LPS，24h）
An inflamma- tory model was established by lipopolysaccharide-stimulated mouse macrophage cell line RAW264.7, and the anti-inflammatory ability of ART was evaluated by ni- trite assay. All cells in this research were cultured in a DEME high sugar medium, in 5% CO2 incubator at 37°C. Mouse macrophages grown in log phase were seeded in a 96-well culture plate at a density of 1×106 cells/well in the DMEM medium. Fresh culture solu- tion containing only 0.5 μg/mL LPS (control group) or 0.5 μg/mL LPS + ART-12% PEG4000 (0.031 mg/mL, 0.063 mg/mL, 0.125 mg/mL, 0.248 mg/mL, 0.496 mg/mL, 0.982 mg/mL) was added to each well, and the culture was continued for 24 hours.
来源文献：Dai F, Bao Y, Li Z, Chen SH, Gao LZ, Liu ZL, Cheng L, Peng Y, Tong XL. Artemisinin is highly soluble in polyethylene Glycol 4000 and such solution has multiple biological effects. Acta Biochim Pol. 2020 May 18;67(2):203-211. doi: 10.18388/abp.2020_5190. PMID: 32421285.
In Vivo
Mice
The LPS-induced acute lung inflammation model
Mice were randomly divided into different groups: saline group, model group (LPS), DAS treatment groups and DAS prevention groups (delivered by either intratracheal instillation or intravenous injection, respectively). The LPS group was treated with 4 μg of LPS per mouse and the saline group was given the same volume of saline. The DAS prevention groups were administered with 25 mg/kg DAS at different time interval (e.g. 1 and 6 h) before LPS challenge. Mice were sacrificed at 6 h after LPS instillation and then the BALF were collected. The DAS treatment groups were administrated with LPS and the DAS (25 mg/kg) were delivered 1 h later. Mice were sacrificed at 6 h after LPS challenge and then the BALF were collected. The BALF was collected 3 times with 0.7 mL of saline each time, and the total volume of ~2 mL was recovered. The BALF samples were centrifuged (4 °C, 1000 rpm, 15 min) and the supernatant was stored at −20 °C for subsequent analysis.
来源文献：Wei-Ya C, Yuan-Song W, Chun-Yu L, Yu-Bin J, Fei-Fei Y, Yong-Hong L. Comparison of pulmonary availability and anti-inflammatory effect of dehydroandrographolide succinate via intratracheal and intravenous administration. Eur J Pharm Sci. 2020 Apr 30;147:105290. doi: 10.1016/j.ejps.2020.105290. Epub 2020 Mar 2. PMID: 32135270.
Mice（Female wild-type C57BL/6 mice， 2 mg/kg/every 3 days LPS, i.p）
In other experiments, mice were serially treated with recombinant CCL2 (rCCL2) (10 µg/kg/every 3 days, i.p.), Toll-like receptor 4 (TLR4) specific ligand (lipopolysaccharide (LPS) -EB Ultrapure, 2 mg/kg/every 3 days, i.p.), LPS (2 mg/kg/every 3 days, i.p.; Jinpan), TLR4 inhibitor (TAK-242, 10 mg/kg/every 3 days, i.p.) or cyclooxygenase-2 (COX-2) inhibitor (Celecoxib, 10 mg/kg/every 3 days, intragastric administration). Polyp or tumour load was determined as the sum of the size of all colon polyps/tumours presented per mouse.
来源文献：Yang Y, Li L, Xu C, Wang Y, Wang Z, Chen M, Jiang Z, Pan J, Yang C, Li X, Song K, Yan J, Xie W, Wu X, Chen Z, Yuan Y, Zheng S, Yan J, Huang J, Qiu F. Cross-talk between the gut microbiota and monocyte-like macrophages mediates an inflammatory response to promote colitis-associated tumourigenesis. Gut. 2020 Oct 29;70(8):1495–506. doi: 10.1136/gutjnl-2020-320777. Epub ahead of print. PMID: 33122176; PMCID: PMC8292576.
Zebrafish larvae（Tg (Lyz:DsRed) transgenic line斑马鱼幼虫）
After exposure to 72 hpf, the Tg (Lyz:DsRed) transgenic line was treated with LPS and cyhalofop-butyl together up to 96 hpf, and then the number of macrophages was observed by neutral red staining to detect the inflammatory response. The tail fins of larvae were cut under a stereomicroscope (Leica S6, Germany) after 96 h of exposure and anesthetized with 0.16% Tricaine.
来源文献：Cheng B, Zou L, Zhang H, Cao Z, Liao X, Shen T, Xiong G, Xiao J, Liu H, Lu H. Effects of cyhalofop-butyl on the developmental toxicity and immunotoxicity in zebrafish (Danio rerio). Chemosphere. 2021 Jan;263:127849. doi: 10.1016/j.chemosphere.2020.127849. Epub 2020 Aug 22. PMID: 33297003.
