Nef inhibits excess expression of COX-2 and iNOS in IL-1β-treated rat chondrocytes. Western blotting results of iNOS and COX-2.
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NF-κB NF-κB1/p50 RelA/p65 RelB c-Rel
Description
Neferine is a major bisbenzylisoquinline alkaloid. Neferine strongly inhibits NF-κB activation.
IC50 & Target[1]
p65
Autophagy
In Vitro
Neferine down regulates hypoxia induced NF-κB p65 nuclear translocation and COX-2 expressions[1]. Neferine reduces high-glucose-induced collagen production and inhibits TGF-β1-Smad, ERK and p38 MAPK signaling activation in cardiac fibroblasts. Cardiac fibroblasts (CFs) are cultured in HG medium with varying concentrations of Neferine (1, 2, or 5 μM). CCK-8 assays are carried out at different time points (24, 48, and 72 h). Compared with normal glucose (NG) and osmotic control (OC) treatments, High glucose (30 mM) treatment significantly increases the proliferation of CFs in a time-dependent manner (P<0.05). High glucose (HG)-induced CF proliferation is markedly attenuated by Neferine treatment at either 2 or 5 μM compared with vehicle treatment. However, 1 μM Neferine does not inhibit HG-induced proliferation of CFs. Therefore, 2 and 5 μM Neferine are used in the remaining experiments[2].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Neferine Related Antibodies
In Vivo
Neferine treatment at both low-dose (60 mg/kg/day by gavage) and high-dose (120 mg/kg/day by gavage) reduces the increment of collagen I, III and TGF-β1 protein expression induced by hyperglycemia[2].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
DMSO : 125 mg/mL (200.07 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
Preparing Stock Solutions
ConcentrationSolventMass
1 mg
5 mg
10 mg
1 mM
1.6006 mL
8.0029 mL
16.0059 mL
5 mM
0.3201 mL
1.6006 mL
3.2012 mL
10 mM
0.1601 mL
0.8003 mL
1.6006 mL
View the Complete Stock Solution Preparation Table
*Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles. Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day. The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.
Protocol 1
Add each solvent one by one: 10% DMSO 40% PEG300 5% Tween-80 45% Saline
This protocol yields a clear solution of ≥ 2.08 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μLDMSO stock solution (20.8 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.
Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
Protocol 2
Add each solvent one by one: 10% DMSO 90% Corn Oil
This protocol yields a clear solution of ≥ 2.08 mg/mL (saturation unknown). If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Taking 1 mL working solution as an example, add 100 μLDMSO stock solution (20.8 mg/mL) to 900 μLCorn oil, and mix evenly.
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:
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Dosing volume (per animal)
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Number of animals
Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
%
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%
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%
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%
Saline
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
The co-solvents required include: DMSO,
. All of co-solvents are available by MedChemExpress (MCE).
, Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Calculation results:
Working solution concentration:
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Method for preparing stock solution:
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The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).
Method for preparing in vivo working solution for animal experiments: Take
μL DMSO stock solution, add
μL .
μL , mix evenly, next add
μL Tween 80, mix evenly, then add
μL Saline.
Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution
If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
[1]. Baskaran R, et al. Neferine prevents NF-κB translocation and protects muscle cells from oxidative stress and apoptosis induced by hypoxia. Biofactors. 2016 Jul 8;42(4):407-17.
[Content Brief]
[2]. Liu X, et al. Neferine inhibits proliferation and collagen synthesis induced by high glucose in cardiac fibroblasts and reduces cardiac fibrosis in diabetic mice. Oncotarget. 2016 Sep 20;7(38):61703-61715.
[Content Brief]
Test cellulaire
[2]
Cardiac fibroblasts (CFs) are isolated from neonatal mouse ventricular tissues. After starvation in serum-free medium for 24 h, CFs are incubated in DMEM containing 5.6 mM glucose (normal glucose; NG), 30 mM D-glucose (HG), 30 mM D-glucose plus 1 μM Neferine, 30 mM D-glucose plus 2 μM Neferine, 30 mM D-glucose plus 5 μM Neferine, and 5.6 mM glucose plus 27.5 mM mannose. Cells are harvested at 24 h, 48 h, and 72h. Cell proliferation is measured with the Cell Counting Kit-8 (CCK-8) and the Cell-LightTM EdU assay[2].
MCE n'a pas confirmé de manière indépendante l'exactitude de ces méthodes. Ils sont pour référence seulement.
Administration animale
[2]
Mice[2] Eight-week-old C57BL/6J male mice are used. Diabetes is induced by intraperitoneal injection of Streptozotocin dissolved in citrate buffer (pH 4.5) at 60 mg/kg body weight for five consecutive days. Control mice are injected with citrate buffer only. Whole blood glucose in mouse tail blood is detected with an Accu-Check Active glucometer. Mice with blood glucose concentrations higher than 18 mM are considered as diabetic animals and used in this study. The animals are randomly divided into four groups of eight animals each. Diabetic mice are divided into three groups: group 1, the diabetic control group (DM); group 2, which receive Neferine at a dose of 60 mg/kg/day (DM-NL); and group 3, which receive Neferine at a dose of 120 mg/kg/day (DM-NH). Neferine is administered twice per day by intragastric gavage for 12 weeks. Equivalent volumes of normal sodium are administered to the normal and DM control groups by gavage. Mice are anaesthetized and sacrificed at the end of the 12-week treatment[2].
MCE n'a pas confirmé de manière indépendante l'exactitude de ces méthodes. Ils sont pour référence seulement.
Références
[1]. Baskaran R, et al. Neferine prevents NF-κB translocation and protects muscle cells from oxidative stress and apoptosis induced by hypoxia. Biofactors. 2016 Jul 8;42(4):407-17.
[Content Brief]
[2]. Liu X, et al. Neferine inhibits proliferation and collagen synthesis induced by high glucose in cardiac fibroblasts and reduces cardiac fibrosis in diabetic mice. Oncotarget. 2016 Sep 20;7(38):61703-61715.
[Content Brief]
[1]. Baskaran R, et al. Neferine prevents NF-κB translocation and protects muscle cells from oxidative stress and apoptosis induced by hypoxia. Biofactors. 2016 Jul 8;42(4):407-17.
[2]. Liu X, et al. Neferine inhibits proliferation and collagen synthesis induced by high glucose in cardiac fibroblasts and reduces cardiac fibrosis in diabetic mice. Oncotarget. 2016 Sep 20;7(38):61703-61715.
Complete Stock Solution Preparation Table
*Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles. Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Optional Solvent
ConcentrationSolventMass
1 mg
5 mg
10 mg
25 mg
DMSO
1 mM
1.6006 mL
8.0029 mL
16.0059 mL
40.0147 mL
5 mM
0.3201 mL
1.6006 mL
3.2012 mL
8.0029 mL
10 mM
0.1601 mL
0.8003 mL
1.6006 mL
4.0015 mL
15 mM
0.1067 mL
0.5335 mL
1.0671 mL
2.6676 mL
20 mM
0.0800 mL
0.4001 mL
0.8003 mL
2.0007 mL
25 mM
0.0640 mL
0.3201 mL
0.6402 mL
1.6006 mL
30 mM
0.0534 mL
0.2668 mL
0.5335 mL
1.3338 mL
40 mM
0.0400 mL
0.2001 mL
0.4001 mL
1.0004 mL
50 mM
0.0320 mL
0.1601 mL
0.3201 mL
0.8003 mL
60 mM
0.0267 mL
0.1334 mL
0.2668 mL
0.6669 mL
80 mM
0.0200 mL
0.1000 mL
0.2001 mL
0.5002 mL
100 mM
0.0160 mL
0.0800 mL
0.1601 mL
0.4001 mL
Neferine Related Classifications
Metabolic DiseaseCancer
Cancer Targeted TherapyCancer ImmunotherapyCancer Metabolism and Metastasis
NF-κBAutophagyApoptosis
NF-κBAutophagyApoptosis
Help & FAQs
Do most proteins show cross-species activity?
Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.