Rat PMVECs (CP-R001) were procured from Procell Life Science & Technology Co., Ltd. (Wuhan, Hubei, China) and cultured in PMVEC-specific complete medium (CM-R001, Procell) at 37 °C with 5% CO 2 . Overexpression plasmid of Sema3A (oe-Sema3A) and the negative control (NC) plasmid were provided by GenePharma Co., Ltd. (Shanghai, China). Well-growing PMVECs were seeded into six-well plates. When the confluence reached around 50%, the cells were transfected with the plasmids using Lipofectamine® 2000 (Thermo Fisher Scientific, Rockford, IL, USA), followed by incubation at 37 °C with 5% CO 2 for 48 h. To induce cell damage, the PMVECs were treated with 1 μg/mL LPS (ST1470, Beyotime Biotechnology Co., Ltd., Shanghai, China) for 4 h. For artificial activation of the ERK/JNK signalling pathways, 4 h after LPS treatment, the cells were treated with 0.1 μM ERK agonist LM22B-10 (HY-104047) or JNK agonist Juglanin (HY-N3442) (both from MedChemExpress, Monmouth Junction, NJ, USA) for 2 h. Overall, the cells were allocated into the following groups: Control (without treatment), LPS, LPS + oe-NC, LPS + oe-Sema3A, LPS+LM22B-10, LPS+LM22B-10+oe-NC, LPS+LM22B-10+oe-Sema3A, LPS+Juglanin, LPS+Juglanin+oe-NC, and LPS+Juglanin+oe-Sema3A.
Lipopolysaccharides (lps)
Manufactured by MedChemExpress
Sourced in United States, China
LPS is a laboratory equipment product designed for filtration and isolation of lipopolysaccharides. It functions as a tool for researchers and scientists to extract and purify LPS from various bacterial sources.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (12 mentions)
Lipopolysaccharides (lps), by Merck Group (10 mentions)
Streptomycin, by Thermo Fisher Scientific (5 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (4 mentions)
Mm 0190r1, by Jiangsu Meimian (3 mentions)
Mm 0180r1, by Jiangsu Meimian (3 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions)
Penicillin, by Thermo Fisher Scientific (3 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions)
Dmem f12, by Thermo Fisher Scientific (2 mentions)
Adenosine triphosphate (atp), by MedChemExpress (2 mentions)
Adenosine triphosphate (atp), by Merck Group (2 mentions)
Tak 242, by MedChemExpress (2 mentions)
Rapamycin, by MedChemExpress (2 mentions)
Rpmi 1640, by Thermo Fisher Scientific (2 mentions)
Raw264, by American Type Culture Collection (2 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (2 mentions)
Propofol, by Merck Group (2 mentions)
Phosphoric acid, by Sinopharm (1 mentions)
A003 1 2, by Nanjing Jiancheng (1 mentions)
43 protocols using lipopolysaccharides (lps)
1
Sema3A Modulates Rat PMVEC Damage
2
PNU-282987 Attenuates LPS/D-GalN-Induced ALF in Mice
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All animal experiments conducted in this study were approved by the Biomedical Ethics Committee of Xi’an Jiaotong University (2022-1559). Male C57BL/6 aged 6 to 8 weeks were obtained from the Laboratory Animal Center of Xi’an Jiaotong University. All mice were housed in a specific pathogen-free environment according to the animal care regulations (12-hour light/12-hour dark cycle) and fed a standard rodent diet. The ALF experimental model was induced by LPS combined with D-GalN. PNU-282987 was used to activate the CAP.
Mice were randomly divided into three groups and the groups were as follows: control: the mice were treated with vehicle; LPS/D-GalN: the mice received intraperitoneally administered LPS (Sigma-Aldrich, 25 μg/kg) in combination with D-galactosamine (D-GalN, Sigma-Aldrich, 700 mg/kg); PNU+LPS/D-GalN: the mice were intraperitoneally administered with PNU-282987 (PNU, 10mg/kg, MedChemExpress) 30 minutes before LPS/D-GalN injections.
Mice were randomly divided into three groups and the groups were as follows: control: the mice were treated with vehicle; LPS/D-GalN: the mice received intraperitoneally administered LPS (Sigma-Aldrich, 25 μg/kg) in combination with D-galactosamine (D-GalN, Sigma-Aldrich, 700 mg/kg); PNU+LPS/D-GalN: the mice were intraperitoneally administered with PNU-282987 (PNU, 10mg/kg, MedChemExpress) 30 minutes before LPS/D-GalN injections.
3
LPS-induced Cardiomyocyte Inflammation
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The mouse cardiomyocyte M6200 cell line was purchased from Zhong Qiao Xin Zhou Biotechnology Co., Ltd (Shanghai, China). The cells were cultured in Dulbecco’s Modified Eagle’s Medium (Thermo Fisher Scientific) containing 10% fetal bovine serum (PAN-Biotech GmbH, Adenbach, Germany) in a humidified incubator (5% CO2 at 37°C). For LPS treatment, M6200 cells were cultured with or without 0.1, 1.0, 10, or 100 mg/L LPS (MedChemExpress Co., NJ, USA) for 24 h, followed by treatment with 125 mg/L rh PLTP (Abmart). Then, the cells were collected for subsequent experiments.
4
Cell Stress Model and Pharmacological Intervention
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The PC-12 cell line was purchased from the National Collection of Authenticated Cell Cultures (China, Shanghai) and the HAPI cell line from BeNa Culture Collection (Beijing, China). PC-12 cells were cultured in RPMI 1640 medium (Gibco, USA) and the HAPI cells in the DMEM/F-12 medium (Gibco). All culture systems were supplemented with 10% fetal bovine serum (FBS) and 1% penicillin–streptomycin (Gibco) under a humidified atmosphere of 5% CO2 at 37 °C.
The cell treatment of the experiments is depicted in Figs.6 and 7 and was as follows: for the LPS/corticosterone + Rg1 group, the cells were pre-treated with LPS (1 μg/mL, to establish an inflammatory stress model) or corticosterone (400 μM, to establish an oxidative stress model) for 2 h, followed by co-incubation with Rg1 (5 μM, 10 μM, 20 μM) for 24 h. For the LPS/corticosterone + shRNA group, the cells were transfected with lentivirus for 48 h and then washed with cold PBS buffer, followed by incubation with LPS or corticosterone for 24 h. corticosterone (HY-B1618) was purchased from MedChemExpress and LPS (L5293) from Sigma-Aldrich.
The cell treatment of the experiments is depicted in Figs.
5
Regulation of Human Mesothelial Cells
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Human peritoneal mesothelial cell line HMrSV5 was purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). HMrSV5 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, Thermofisher Scientific, Waltham, MA, USA) with adding 10% fetal bovine serum (FBS) (Gibco, Thermofisher Scientific, Waltham, MA, USA), 100 ul/ml streptomycin and 100 U/ml penicillin (Life Technologies, Carlsbad, CA, USA). The cells were incubated at 37°C humidified atmosphere with 5% CO2 until 50–70% confluence before used. Cells were transferred to serum-free DMEM medium 12 h prior to each experiment. Cells in the control group were maintained in serum-free DMEM medium. For the experimental groups, HMrSV5 cells were treated with high glucose (25 μM) and lipopolysaccharide (LPS0, 910 μg/ml) for 24 h. As for the role of 1,25(OH)2D3, HMrSV5 cells were pre-incubated with 1,25(OH)2D3 (10−6 mol/L) for 2 h and subsequently treated with high glucose (25 μM) and LPS (10 μg/ml) for 24 h. Wnt agonist 1, 0.7 μM for 12 h (MedChem Express, Monmouth Junction, NJ, USA) was also used, as previously described [23 (link)]. Glucose, LPS, and 1,25(OH)2D3 were purchased from Sigma-Aldrich (St. Louis, MO, USA). The concentrations of glucose and LPS were determined from the findings of preliminary experiments.
6
Investigating Macrophage Response to LPS and PF Treatment
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The RAW 264.7 (2 × 105) mouse monocyte line (ATCC TIB-71; American Type Culture Collection, Manassas, VA) was used as the macrophage line. The cells were cultured in 35-mm dishes with high-glucose Dulbecco modified Eagle medium (Thermo Fisher Scientific, Waltham, MA) and 10% fetal bovine serum (Gibco). When 70% confluence was reached (generation 3–4), randomization of the cells into the following three groups was performed for subsequent experiments: control, LPS model (LPS group), and PF+LPS groups. LPS (Sigma-Aldrich, St. Louis, MO, USA) was added to the latter two groups at the same time point, and cells in the PF+LPS group were pretreated with 10−5 mol/L PF (MedChemExpress, USA) for 4 hour and cultured for 12 hour after the addition of 1 μg/mL LPS. An equivalent volume of PBS was added to the control group. The cells were cultured in an incubator at 37°C with 5% CO2 prior to further experimentation.
7
Polycaprolactone and PLCL Biomaterials
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Polycaprolactone (PCL) 80 K (Mw =80 kDa) and poly (L-lactide-co-caprolactone) (PLCL) with an L-lactic acid/ε-caprolactone ratio of 75:50 (MW =385 kDa) were from Patos Biomaterial Co., Ltd. (Tianjin, China). Chromatographic (chromatographical purity) was bought from Fisher Scientific (Leicestershire, UK). Phosphoric acid was purchased from Sinopharm Group Chemical Reagent Co., Ltd. Methylprednisolone sodium succinate (MPSS, purity ≥97%) was purchased from Shanghai Haling Biotechnology Co., Ltd., Shanghai, China. Lipopolysaccharides (LPS) were purchased from Medchemexpress, Shanghai, China. All other chemicals were of analytical grade and used as received. All the deionized water was homemade. The Gore Excluder Iliac Branch Endoprosthesis was kindly provided by the Gore Company (IBE, W. L. Gore and Associates, Flagstaff, AZ, USA).
8
Anti-oxidative and Anti-inflammatory Assays
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Assay kits for SOD, MDA, CAT, GPx, and MPO were obtained from Jiancheng Bioengineering Institute (A001-3-2, A003-1-2, A007-1-1, H545-1-1, and A044-1-1) (Nanjing, China). ELISA assays for PGI, PGII, iNOS, NO, PGE2, TNF-α, IL-1β, and IL-6 were obtained from MEIMIAN (MM-70280R1, MM-70274R1, MM-0889R1, MM-70810R2, MM-0068R1, MM-0180R1, MM-0047R2, and MM-0190R1). Rabbit anti-NF-κBp65 conjugated antibody (AF5006), rabbit anti-p-NF-κBp65 conjugated antibody (AF2006), and mouse anti-β-actin antibody (T0022) were obtained from Affinity. Lipopolysaccharides (LPS) (batch number HY-D1056) were obtained from MedChemExpress.
9
PET Polymer Characterization and Macrophage Activation
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Our analyses utilized two types of PET sourced from Dongguan Zhangmutou, China: (i) polydisperse PET powders (with a crystallinity >25% and particle size <400 μm), and (ii) amorphous PET film (with a crystallinity of 7% and a thickness of 250 μm). Terephthalic acid (TPA) (CAS: 100-20-1, product code: P816020-100 g), mono(2-hydroxyethyl) terephthalic acid (MHET) (CAS: 1137-99-1, product code: H909109-100 mg), and dimethyl sulfoxide (DMSO) (CAS: 67-68-5, product code: D806645-500 mL) were procured from Macklin, Shanghai, China. TPA and MHET were dissolved in DMSO, and the medium containing the same concentration of DMSO was used as a vehicle control. Macrophage colony-stimulating factor (M-CSF) (product code: HY-P7085), lipopolysaccharides (LPS) (product code: HY-D1056), and murine interleukin-4 (IL-4) (product code: HY-P70644) were obtained from MedChemExpress, Princeton, NJ, USA.
10
Detailed Antibody Protocol for Research
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The primary antibodies used in this study are all listed in Table S1 . The mouse immunoglobulin G (IgG) and rabbit IgG were purchased from Sigma. The fluorescein and horseradish peroxidase (HRP) labelled secondary antibodies were obtained from Li‐Cor Biosciences and Proteintech Group, respectively. The Alexa Fluor 647‐, 594‐ and 488‐conjugated secondary antibodies were from Thermo Fisher Scientific. The enhanced chemiluminescent substrate was purchased from Biosharp. Neofect DNA transfection reagent was from Neofect Biological Technology Co. Ltd. TPOP146, lipopolysaccharides (LPS) and nigericin were purchased from MedChemExpress. L‐Moses hydrochloride (L‐45) was from GlpBio. The protein marker, dulbecco's modified eagle medium (DMEM), high glucose, DMEM/F12, foetal bovine serum (FBS) and Lipofectamine 3000 were from Invitrogen and Gibco (Thermo Fisher Scientific). Protein A/G agarose was from Smart Lifesciences. All other chemical reagents were standard commercial products with analytical reagent grade.
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