LPS (Sigma-Aldrich, St. Louis, MO, USA) was used to induce ALI model [17 ] and 72 mice were grouped into six groups with 12 mice in each group: (1) Blank group (no treatment); (2) Phosphate buffered saline (PBS) group (mice were administered with PBS through nose); (3) LPS group (mice were administered with LPS through nose); (4) LPS + normal saline (NS) group (mice were administered with LPS through nose and injected intraperitoneally with normal saline); (5) LPS + 0.3 Tunicamycin (TM) group (mice were administered with LPS through nose and injected intraperitoneally with 0.3 mg/kg TM); (6) LPS + 3.0 TM group (mice were administered with LPS through nose and injected intraperitoneally with 3.0 mg/kg TM). Mice in the LPS + NS group, LPS + 0.3 TM group and LPS + 3.0 TM group were daily intraperitoneally injected with NS or TM (Sigma-Aldrich, St. Louis, MO, USA) 3 d before model establishment. One hour after the final time of injection, mice were anesthetized using pentobarbital sodium (Sigma-Aldrich, St. Louis, MO, USA). Except for mice in the blank group and the PBS group, mice were administered with 0.5 mg/kg LPS, while mice in the PBS group were administered with 0.5 mg/kg PBS. About 12 h later, mice were euthanized and the sample was gathered.
Lipopolysaccharides (lps)
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The LPS laboratory equipment is a high-precision device used for various applications in scientific research and laboratory settings. It is designed to accurately measure and monitor specific parameters essential for various experimental procedures. The core function of the LPS is to provide reliable and consistent data collection, ensuring the integrity of research results. No further details or interpretations can be provided while maintaining an unbiased and factual approach.
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Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (645 mentions)
Interleukin 4 (il 4), by Thermo Fisher Scientific (293 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (248 mentions)
Ifn γ, by Thermo Fisher Scientific (245 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (240 mentions)
Penicillin, by Thermo Fisher Scientific (194 mentions)
Streptomycin, by Thermo Fisher Scientific (187 mentions)
Phorbol myristate acetate (pma), by Merck Group (172 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (166 mentions)
Rpmi 1640, by Thermo Fisher Scientific (140 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (137 mentions)
Interleukin 4 (il 4), by R&D Systems (129 mentions)
Adenosine triphosphate (atp), by Merck Group (110 mentions)
Ifn γ, by R&D Systems (92 mentions)
Ifn γ, by Merck Group (90 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (89 mentions)
Lipopolysaccharides (lps), by Thermo Fisher Scientific (88 mentions)
Dexamethasone (dex), by Merck Group (85 mentions)
Gm csf, by Thermo Fisher Scientific (74 mentions)
Griess reagent, by Merck Group (71 mentions)
7 203 protocols using lipopolysaccharides (lps)
1
Modeling Acute Lung Injury in Mice
2
Nthy-ori 3–1 Cell Line Treatments
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The Nthy-ori 3–1 cell line was obtained from the China Center Type Culture Collection (Wuhan, China). Cells were cultured in RPMI 1640 medium supplemented with 2 mM glutamine and 10% fetal bovine serum in a humidified atmosphere containing 5% CO2. Cells between passage 5 and passage 12 were used for all experiments. The cells were divided into the following six groups:
A, the normal control (Con) group, which received medium only;
B, the Con +PV group, which received 5 μmol/L PV;
C, the lipopolysaccharide (LPS)+ NaI group, which received 10 µg/mL LPS (Sigma) and 100 mg/L NaI;
D, the LPS+NaI +PV group, which received 10 µg/mL LPS + 1×10−2 mmol/L NaI + 5 μmol/L PV;
E, the LPS + NaI+ EP group, which received 10 µg/mL LPS+1×10−2 mmol/L NaI + 5 mmol/L EP; and
F, the LPS+NaI+PV+EP group, which received 10 µg/mL LPS +1×10−2 mmol/L NaI + 5 μmol/L PV+ 5 mmol/L EP.
A, the normal control (Con) group, which received medium only;
B, the Con +PV group, which received 5 μmol/L PV;
C, the lipopolysaccharide (LPS)+ NaI group, which received 10 µg/mL LPS (Sigma) and 100 mg/L NaI;
D, the LPS+NaI +PV group, which received 10 µg/mL LPS + 1×10−2 mmol/L NaI + 5 μmol/L PV;
E, the LPS + NaI+ EP group, which received 10 µg/mL LPS+1×10−2 mmol/L NaI + 5 mmol/L EP; and
F, the LPS+NaI+PV+EP group, which received 10 µg/mL LPS +1×10−2 mmol/L NaI + 5 μmol/L PV+ 5 mmol/L EP.
3
Macrophage-Mediated Endothelial Protection
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Macrophages from M‐Wt, M‐Vec, and IL10‐eM groups were cultured with a culture medium containing FBS in 10 cm dishes, respectively. When reached 80–90% confluency, the culture medium was replaced with Dulbecco's modified Eagle medium (DMEM) without FBS and antibiotics and cells continued to be cultured for 24 h. Thereafter, the supernatants from different groups were collected and frozen at −80 °C after centrifuging at 1000 g for 10 min. In a noncontact coculture system, C166 was treated with LPS to simulate septic endothelial injury, and then, supernatants from different macrophages were added to investigate the effect on endothelial dysfunction. The detailed treatment methods and experimental group settings are shown as below: (a) Control (CTL) group: DMEM cultured for 24 h, (b) LPS group: DMEM containing 1 μg·mL−1 LPS (Sigma, USA) was treated for 24 h, (c) M‐Wt + LPS group: M‐Wt supernatant was pretreated for 30 min and then added 1 μg·mL−1 LPS for 24 h, (d) M‐Vec + LPS group: M‐Vec supernatant was pretreated for 30 min and then added 1 μg·mL−1 LPS for 24 h, (e) IL10‐eM + LPS group: IL10‐eM supernatant was pretreated for 30 min and then added 1 μg·mL−1 LPS for 24 h.
4
Dexamethasone Modulation of LPS-Stimulated Alveolar Macrophages
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After incubating for 24 hours, AMs were randomly divided into six groups: 1) control group, AMs were incubated with PBS; 2) lipopolysaccharide (LPS; Sigma-Aldrich, St. Louis, MO, USA) group, AMs were stimulated with LPS (1 μg/mL); 3) LPS +10−4 M Dex group (Sigma-Aldrich); 4) LPS +10−5 M Dex group; 5) LPS +10−6 M Dex group; 6) LPS +10−7 M Dex group; AMs were incubated with LPS for 24 hours and then stimulated with different doses of Dex (10−4, 10−5, 10−6, or 10−7 M) for another 6, 24, or 48 hours. AMs were then cultured for assay-specific periods of time in a humidified chamber containing 5% CO2 at 37°C. Cell-free supernatants were kept at −80°C for later analysis.
5
Monocyte-like Cells: LPS, E3330, and MX
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U937 monocyte-like cells, derived from patients with histiocytic lymphoma (ATCC® CRL1593.2), were cultured in Gibco RPMI-1640 medium (Thermo Fisher Scientific Inc., Waltham, MA, United States) supplemented with 44 mM sodium bicarbonate (Sigma-Aldrich, St. Louis, MO, United States), Gibco 10% fetal bovine serum (Thermo Fisher Scientific Inc.), and 1% Penicillin-Streptomycin antibiotic solution (Sigma-Aldrich) in a humidified incubator at 37°C and a 5% CO2 atmosphere unless stated otherwise. Cells (5 × 105) in 3 mL of medium were stimulated with 1 μg/mL LPS (Cat. No: L2654; Sigma-Aldrich) for 24 h, and then incubated with 100 μM E3330 (≥98% pure; Sigma-Aldrich) or 6 mM methoxyamine (MX) for 4 h. The cells were grouped as follows: unstimulated (control), LPS-stimulated (LPS), LPS + 100 μM E3330 (LPS + E3330), and LPS + 6 mM MX (LPS + MX). We ensured that the cell lines used in these experiments were free of mycoplasma infection.
6
Rat Model of LPS-Induced Inflammation
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Twenty male Sprague-Dawley rats (7–8 weeks old) were purchased from Shanghai SLAC Laboratory Animal Co., Ltd. (Shanghai, China). All the rats were kept in pathogen-free housing under a 12-h light/dark cycle with free access to food and sterile water during the research procedures. The rats were anesthetized with 3% sodium pentobarbital (150 mg/kg), and randomly divided into four groups (five rats per group): control (K group): intraperitoneal injection of physiological saline; LPS (L group): intraperitoneal injection of 200 µL of LPS (5 mg/kg; Sigma, St. Louis, MO, USA) dissolved in physiological saline; LPS + dexamethasone (LPS + Dexa group, positive control): intragastric administration of Dexa (1.5 mg/kg) (Sigma) 1 h before intraperitoneal injection of LPS (5 mg/kg); LPS + Res (R group in sequencing data): intragastric administration of Res (0.5 mg/kg) 1 h before intraperitoneal injection of LPS (5 mg/kg). At 6 h after LPS treatment, the rats were euthanized by sodium pentobarbital injection (200 mg/kg). All experimental procedures were approved by the ethics committee of Shanghai Pulmonary Hospital (approval No. K21-025), in compliance with National Institutes of Health Guidelines for the Care and Use of Laboratory Animals.
7
Effects of pBD114 on LPS-induced Inflammation
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A mouse assay was performed to investigate the effects of pBD114 on the inflammatory response in the animals; a total of 40 C57BL/6 mice (7 weeks) were randomly allocated to four groups: CON, treated without pBD114 and LPS; PBD114, treated with 10 mg/kg pBD114 and without LPS (Sigma-Aldrich, St Louis, MO, USA); LPS, treated without pBD114 and with 1 mg/kg LPS; and PBD114LPS, treated with 1 mg/kg pBD114 and with 1 mg/kg LPS. pBD114 and LPS were intraperitoneally injected using a 1 mL insulin syringe (Braun, Melsungen, Germany) within 200 μL PBS. After 12 h, all mice were sacrificed using carbon dioxide anesthesia, their livers and serum were collected for analysis, and their weight was measured.
8
Evaluating Anti-inflammatory Compounds in LPS-Induced Mouse Model
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Animal studies were conducted according to protocols approved by Institutional Animal Care and Use Committee of China Pharmaceutical University. All animals were appropriately used in a scientifically valid and ethical manner. After treatment with regular drinking water for 2 days for adaptation, female C57BL/6 mice (6–8 weeks of age, weighing 18–20 g) were randomized into eight groups: (A) Control group (n = 3); (B) LPS (Sigma-Aldrich, St. Louis, no. L4130) model group (300 μg/kg/day, n = 8); (C) LPS (300 μg/kg/day) + pro2 low-dose (10 mg/kg/day) group (n = 8); (D) LPS (300 μg/kg/day) + pro2 high-dose (40 mg/kg/day) group (n = 8); (E) LPS (300 μg/kg/day) + parent compound 2 high-dose (40 mg/kg/day) group (n = 8); (F) LPS (300 μg/kg/day) + dexamethasone low-dose (10 mg/kg/day) group (n = 8); (G) pro2 high-dose (40 mg/kg/day) group (n = 3); (H) parent compound 2 high-dose (40 mg/kg/day) group (n = 3).
Animals in control and (G, H) groups received a single IP injection containing 500 μL of saline (day −3, −2, −1). All LPS-challenged mice received a single IP injection containing 500 μL of LPS (day −3, −2, −1). Four hours after the injection, mice were pretreated with compound (ig, in 500 μL of saline, day −3, −2, −1). Animals were sacrificed 24 h after the last dose of compound and sera were collected (day 0). The level of the cytokines in sera was measured using ELISA kits.
Animals in control and (G, H) groups received a single IP injection containing 500 μL of saline (day −3, −2, −1). All LPS-challenged mice received a single IP injection containing 500 μL of LPS (day −3, −2, −1). Four hours after the injection, mice were pretreated with compound (ig, in 500 μL of saline, day −3, −2, −1). Animals were sacrificed 24 h after the last dose of compound and sera were collected (day 0). The level of the cytokines in sera was measured using ELISA kits.
9
Adiponectin and LPS Stimulation on HFLS Cells
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The cells were stimulated for 24 and 72 h with optimal doses of LPS (Sigma) and adiponectin (Peprotech, Inc., Cranbury, NJ, USA), respectively, in different variants: (1) LPS1000 ng/mL, (2) adiponectin 250 ng/mL, (3) adiponectin 1000 ng/mL, (4) adiponectin 250 ng/mL + LPS1000 ng/mL, and (5) adiponectin 1000 ng/mL + LPS1000 ng/mL. HFLS Basal Medium with BSA 0.5% (Sigma) was used as a negative control. The samples were performed in triplicate. After the time points, the cell pellets were used for total RNA isolation and reverse transcriptase-polymerase chain reaction (RT-PCR) analysis. LPS was derived from E. coli bacteria (lipopolysaccharides from Escherichia coli O55: B5) and was purchased from Sigma-Aldrich, St. Louis, MO, USA (Cat # L6529).
10
CLA Modulates Ruminal Epithelial Inflammation
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The immortalized ruminal epithelial cell line RECs was cultured in DMEM (Gibco, New York, USA) containing 2% FBS, 1% penicillin/streptomycin and 1% epithelial cell additive at 37 °C with 5% CO2, as described previously [29 (link)]. When REC monolayers reached 70–80% confluency, the cells were treated without (control group, CON) or with 0.1 μg/mL LPS (from Escherichia coli O111:B4, Sigma-Aldrich, Shanghai, China) for 3 h [29 (link)] after pretreatment with 50 or 100 μM cis-9,trans-11- (c-9,t-11-CLA+LPS group, c-9,t-11-CLA+LPS) or trans-10,cis-12-CLA (t-10,c-12-CLA+LPS group, t-10,c-12-CLA+LPS) (purity ≥ 96.0%, Sigma-Aldrich, Shanghai, China) for 24 h or not (LPS group, LPS). Six treatments were contained and each treatment had 4 replicates.
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