Journal: Mediators of Inflammation

Article Title: Levistilide A Ameliorates NLRP3 Expression Involving the Syk-p38/JNK Pathway and Peripheral Obliterans in Rats

doi: 10.1155/2018/7304096

Figure Lengend Snippet: LA inhibited the mRNA levels of LPS-induced proinflammatory mediators and inhibited the adhesion of monocytes to HUVECs. The molecular structure of LA (a). HUVECs were pretreated with 50 μ M of LA for 60 min before treatment with LPS (100 ng/mL) (b) for 6 h. Expression of IL-1 β , TNF- α , IL-32 β , MCP-1, CCL-2, and VCAM-1 mRNA levels were measured using RT-PCR. HUVECs were pretreated with a blocking mouse antihuman monoclonal antibody (E1/6) against VCAM-1 or various doses of LA for 60 min before treatment with LPS. U937 mononuclear cells adhered to LPS-stimulated HUVECs were observed through microscopic images (c). Data represent the mean ± SD of at least three independent experiments, and each experiment was performed in triplicate. # p < 0.05 versus control; ∗ p < 0.05 versus LPS; ∗∗ p < 0.01 versus LPS; ∗∗∗ p < 0.001 versus LPS.

Article Snippet: Monoclonal mouse anti-GAPDH (glyceraldehyde 3-phosphate dehydrogenase) and horseradish peroxidase- (HRP-) conjugated antibody (KC-5G5) were purchased from KangChen Bio-tech Inc. (Shanghai, China). p-Syk (Tyr535/526), ERK (4695S), p-ERK (4370S), p38 (8690S), p-p38 (4511S), JNK (9258S), p-JNK (4668S), and IL-1 β (12703S) antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Blocking Assay

Journal: Mediators of Inflammation

Article Title: Levistilide A Ameliorates NLRP3 Expression Involving the Syk-p38/JNK Pathway and Peripheral Obliterans in Rats

doi: 10.1155/2018/7304096

Figure Lengend Snippet: LA suppressed NLRP3 gene and downstream related protein expression. HUVECs were treated with 50 μ M of LA for 1 h, and then LPS was stimulated for 24 h. The NLRP3 mRNA level (a) was analyzed using qRT-PCR. LA and p38/JNK inhibitors (SB203580 and SP600125) were treated with HUVECs for 60 min before LPS stimulation for 24 h. NLRP3-associated proteins IL-1 β and IL-18 were detected using immunoblotting (b)(c). Data represent the mean ± SD of at least three independent experiments, and each experiment was performed in triplicate. # p < 0.05 versus control; ∗ p < 0.05 ∗∗ p < 0.01 versus LPS.

Article Snippet: Monoclonal mouse anti-GAPDH (glyceraldehyde 3-phosphate dehydrogenase) and horseradish peroxidase- (HRP-) conjugated antibody (KC-5G5) were purchased from KangChen Bio-tech Inc. (Shanghai, China). p-Syk (Tyr535/526), ERK (4695S), p-ERK (4370S), p38 (8690S), p-p38 (4511S), JNK (9258S), p-JNK (4668S), and IL-1 β (12703S) antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Mediators of Inflammation

Article Title: Levistilide A Ameliorates NLRP3 Expression Involving the Syk-p38/JNK Pathway and Peripheral Obliterans in Rats

doi: 10.1155/2018/7304096

Figure Lengend Snippet: LA reduced the rats' inflammatory factor levels. Wistar rat serum IL-1 β /IL-6/CRP content was detected using an ELISA (a–c). Lower limb arterial NLRP3 mRNA levels in rats (d). # p < 0.05 versus negative control group; ∗ p < 0.05 and ∗∗∗ p < 0.001 versus positive control group.

Article Snippet: Monoclonal mouse anti-GAPDH (glyceraldehyde 3-phosphate dehydrogenase) and horseradish peroxidase- (HRP-) conjugated antibody (KC-5G5) were purchased from KangChen Bio-tech Inc. (Shanghai, China). p-Syk (Tyr535/526), ERK (4695S), p-ERK (4370S), p38 (8690S), p-p38 (4511S), JNK (9258S), p-JNK (4668S), and IL-1 β (12703S) antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Negative Control, Positive Control

Journal: Mediators of Inflammation

Article Title: Levistilide A Ameliorates NLRP3 Expression Involving the Syk-p38/JNK Pathway and Peripheral Obliterans in Rats

doi: 10.1155/2018/7304096

Figure Lengend Snippet: LA anti-inflammatory mechanism diagram. LA inhibited the activity of p38/JNK by decreasing Syk phosphorylation, thereby inhibiting the downstream NLRP3 inflammasome gene expression and the secretion of its related proteins (IL-1 β and IL-18).

Article Snippet: Monoclonal mouse anti-GAPDH (glyceraldehyde 3-phosphate dehydrogenase) and horseradish peroxidase- (HRP-) conjugated antibody (KC-5G5) were purchased from KangChen Bio-tech Inc. (Shanghai, China). p-Syk (Tyr535/526), ERK (4695S), p-ERK (4370S), p38 (8690S), p-p38 (4511S), JNK (9258S), p-JNK (4668S), and IL-1 β (12703S) antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Activity Assay, Expressing

Journal: Neural Plasticity

Article Title: Electroacupuncture Promotes the Survival of the Grafted Human MGE Neural Progenitors in Rats with Cerebral Ischemia by Promoting Angiogenesis and Inhibiting Inflammation

doi: 10.1155/2021/4894881

Figure Lengend Snippet: Effects of MGE neural progenitors and EA treatment on the expression of TNF- α and IL-1 β in the hippocampus were evaluated by Western blot 14 days after grafting. (a) Gel electrophoresis of TNF- α and IL-1 β . (b, c) Densitometric analysis of TNF- α (one-way ANOVA, F = 91.711, P < 0.001) and IL-1 β (one-way ANOVA, F = 57.059, P < 0.001). Values are mean ± SEM ( N = 5 rats/per group). ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001.

Article Snippet: After electrophoresis, it was transferred onto polyvinylidene difluoride (PVDF) membranes, blocked in milk, and incubated at 4°C overnight with primary antibodies: rabbit anti-VEGF (Affinity, AF5131, 1 : 1000), rabbit anti-TNF- α (CST, 8184S, 1 : 1000), and rabbit anti-IL-1 β (CST, 12703S, 1 : 1000), followed by incubation with appropriate horseradish peroxidase-conjugated secondary antibodies.

Techniques: Expressing, Western Blot, Nucleic Acid Electrophoresis

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Interleukin-37 Attenuates Lipopolysaccharide (LPS)-Induced Neonatal Acute Respiratory Distress Syndrome in Young Mice via Inhibition of Inflammation and Cell Apoptosis

doi: 10.12659/MSM.920365

Figure Lengend Snippet: IL-37 attenuated excessive inflammatory cytokines release and lung pathological manifestations in an LPS-induced neonatal ARDS model. ( A ) Tumor necrosis factor-α (TNF-α), ( B ) interleukin-1β (IL-1β), ( C ) IL-8, and ( D ) monocyte chemotactic protein 1 (MCP-1) levels were detected with ELISA kits. ( E ) Representative images of hematoxylin and eosin (HE) staining lung tissues from different groups (×200 magnification). The data are expressed as means±SD from 3 independent experiments; *** P <0.001 vs. Control. ### P <0.001 vs. LPS.

Article Snippet: Rabbit anti-ICAM-1 antibodies (1: 1000), rabbit anti-CXCR4 antibodies (1: 1000), rabbit anti-SDF-1 antibodies (1: 1000), rabbit anti-Bcl2 antibodies (1: 1000), rabbit anti-bax antibodies (1: 1000), rabbit anti-cleaved caspase3 antibodies (1: 500), rabbit anti-NLRP3 antibodies (1: 1000), rabbit anti-ASC antibodies (1: 1000), rabbit anti-pro-IL-1β antibodies (1: 1000), rabbit anti-IL-1β antibodies (1: 1000), rabbit anti-pro-caspase1 antibodies (1: 1000), and rabbit anti-caspase1 antibodies (1: 1000) were purchased from Cell Signaling Technology (Danvers, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Staining

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Interleukin-37 Attenuates Lipopolysaccharide (LPS)-Induced Neonatal Acute Respiratory Distress Syndrome in Young Mice via Inhibition of Inflammation and Cell Apoptosis

doi: 10.12659/MSM.920365

Figure Lengend Snippet: IL-37 inactivated NLRP3 inflammasome pathway in LPS-induced neonatal ARDS model induced by LPS. ( A ) The expression levels of NLRP3, ASC, IL-1β, pro-IL-1β, pro-caspase-1, and caspase-1 were measured by Western blotting at 12 h. ( B ) The expression levels of NLRP3, ASC, IL-1β, pro-IL-1β, pro-caspase-1, and caspase-1 were measured by Western blotting at 24 h. ( C ) NLRP3 expression level was analyzed by immunohistochemistry (×200 magnification). The data are expressed as means±SD from 3 independent experiments; ** P <0.01, *** P <0.001 vs. Control. # P <0.05, ### P <0.001 vs. LPS.

Article Snippet: Rabbit anti-ICAM-1 antibodies (1: 1000), rabbit anti-CXCR4 antibodies (1: 1000), rabbit anti-SDF-1 antibodies (1: 1000), rabbit anti-Bcl2 antibodies (1: 1000), rabbit anti-bax antibodies (1: 1000), rabbit anti-cleaved caspase3 antibodies (1: 500), rabbit anti-NLRP3 antibodies (1: 1000), rabbit anti-ASC antibodies (1: 1000), rabbit anti-pro-IL-1β antibodies (1: 1000), rabbit anti-IL-1β antibodies (1: 1000), rabbit anti-pro-caspase1 antibodies (1: 1000), and rabbit anti-caspase1 antibodies (1: 1000) were purchased from Cell Signaling Technology (Danvers, USA).

Techniques: Expressing, Western Blot, Immunohistochemistry

Journal: Journal of Ophthalmology

Article Title: Low-Dose Mitomycin C Decreases the Postoperative Recurrence Rate of Pterygium by Perturbing NLRP3 Inflammatory Signalling Pathway and Suppressing the Expression of Inflammatory Factors

doi: 10.1155/2019/9472782

Figure Lengend Snippet: MMC suppresses the activation of NLRP3/Caspase-1 pathway in pterygium tissues. (a) NLRP3 expression in typical tissue sections from primary pterygium patients with or without MMC injection was determined by immuno-histochemistry analysis. Data were demonstrated as corresponding images (left panels) and staining score of NLRP3 with means ± SD (right panels). ∗∗ P < 0.01. (b) Comparison of NLRP3 protein levels between group I and group II in primary pterygium patients was carried out by western blotting analysis. The expression levels of NLRP3 were normalized to those of β -actin, the horizontal line represents the median value, and the error bars indicate the SEM. ∗∗ P < 0.01. (c) The positive relation between fibroblasts numbers and NLRP3 expression was determined by Spearman's correlation analysis. β -Actin protein was used as internal reference (Spearman's R = 0.663, ∗ P < 0.05). (d, e) The protein expression of Caspase-1 and Cleaved Caspase-1 in group I and group II patients was determined by western blotting analysis. Data are shown as representatives (d) or relative densities compared with β -actin from three independent experiments with means ± SD (e) ∗∗ P < 0.01 and # P > 0.01. (f) The expression of IL-1 β in primary pterygium patients from group I and group II was determined by immuno-histochemistry analysis. Data are represented as representatives (left panels) or means ± SD of score staining of IL-1 β (right panels). ∗∗ P < 0.01. (g) Relative gray values contrasted with β -actin assessed by western blotting analysis from three independent experiments with means ± SD. ∗∗ P < 0.01. (h) The mRNA expression of IL-18 in pterygium tissues from group I and group II was measured by real-time PCR analysis. Data are represented as the relative expression towards GAPDH of means ± SD in three independent experiments. ∗∗ P < 0.01. Normal corneal tissues were used as controls in all of above experiments.

Article Snippet: Rabbit antihuman NLRP3(#13158), rabbit antihuman IL-1 β (#12703), rabbit Caspase-1(#3866), rabbit antihuman Cleaved Caspase-1(#4199), rabbit antihuman TGF- β 1(#5154), rabbit antihuman VEGF(#2445), and rabbit antihuman IL-6(#12153), rabbit antihuman IL-8(#94853), rabbit antihuman TNF- α (#8184) monoclonal antibodies (mAbs), and horseradish peroxidase (HRP)-conjugated secondary(#7075) mAbs were purchased from Cell Signaling Technology.

Techniques: Activation Assay, Expressing, Injection, Immunohistochemistry, Staining, Western Blot, Real-time Polymerase Chain Reaction

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Cichoric Acid Ameliorates Monosodium Urate-Induced Inflammatory Response by Reducing NLRP3 Inflammasome Activation via Inhibition of NF- k B Signaling Pathway

doi: 10.1155/2021/8868527

Figure Lengend Snippet: Effect of CA on the mRNA levels of iNOS and CD86 in the THP-M cells. The expression levels of iNOS and CD86 were detected by RT-PCR. The mRNA levels of iNOS and CD86 were normalized to MUS (a). Inhibitory effects of CA on IL-1 β and TNF- α secretion in the THP-M cells. The culture medium was collected, and the levels of IL-1 β and TNF- α were determined by ELISA (b). The results are presented as mean ± SD ( n = 3). ∗ p < 0.05, ∗∗ p < 0.01 vs. the MUS group.

Article Snippet: The specifics of the other materials used in the study are as follows: COX-1 Inhibitor Screening Kit (Fluorometric) (BioVision, Inc., Mountain View, CA, USA); COX-2 Inhibitor Screening Kit (Beyotime Biotech Co., Ltd., China); PVDF (0.45 μ m) (Millipore, Schwalbach, Germany); precolor protein marker (Green BioResearch, LA, USA); fuchsia, Tween 20, acrylamide, and sodium dodecyl sulfate (Solon, OH, USA); protein lysate (RIPA) (Beyotime Biotechnology, Shanghai, China); western blotting membrane regeneration solution (C500031, Sangon Biotech, Shanghai, China); fetal bovine serum (FBS) (GIBCO, NY, USA); RPM1-1640 culture medium (GIBCO, NY, USA); trypsin (Gibco, Grand Island, NY, USA); phorbol myristate acetate (PMA) and penicillin (Sigma-Aldrich, St. Louis, MO); TNF- α ELISA kit (DTA00D) and IL-1 β ELISA kit (DLB50) (R&D Systems, Minnesota, USA); anti-CD86 antibody (ab77276), anti-TNF- α antibody (ab269282), anti-NLRP3 antibody (ab263899), PKA kinase activity assay kit (ab139435), Prostaglandin E2 ELISA Kit (ab133021), and cAMP assay kit (Competitive ELISA) (ab234585) (Abcam, Cambridge, UK); anti-IL-1 β antibody (12703), anti-phos-p65 (Ser536) antibody (3033), anti-p65 antibody (8242), anti-phos-I κ B α (Ser32) antibody (2859), anti-I κ B α antibody (9242), anti-COX-2 antibody (12282) (CST, Boston, USA), and TRIzol (Invitrogen); reverse transcription kit (1708843) and SsoFast EvaGreen Supermix (1725200) (Bio-Rad Laboratories, California, USA); primer (Sangon Biotech Co., Ltd. Shanghai, China); BD Cytofix/Cytoperm™ Fixation/Permeabilization Kit (554714) (BD, New Jersey, USA); ionomycin (SQ23377) (MCE, Shanghai, China); MSU (Invivogen, France); Berthold LB941 microporous plate-type multifunctional enzyme labeling instrument; flow cytometry (Beckman DxFLEX, USA); real-time fluorescence quantitative PCR instrument (ABI 7500, ABI, USA).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Cichoric Acid Ameliorates Monosodium Urate-Induced Inflammatory Response by Reducing NLRP3 Inflammasome Activation via Inhibition of NF- k B Signaling Pathway

doi: 10.1155/2021/8868527

Figure Lengend Snippet: Relative levels of phosphorylation of multiple proteins in the THP-M cells. The protein levels of p65, IL-1 β , pro-IL-1 β , I κ B α , and NLRP3 and the levels of phosphorylated p65 (Ser536) and p-I κ B α (Ser32) from different groups were analyzed by western blotting (a). The protein levels of phos-p65 (Ser536) (b), phos-I κ B α (Ser32) (c), I κ B α (d), IL-1 β (e), and NLRP3 (f) were normalized to MUS. The results are presented as mean ± SD ( n = 3). ∗∗ p < 0.01 vs. the MSU group.

Article Snippet: The specifics of the other materials used in the study are as follows: COX-1 Inhibitor Screening Kit (Fluorometric) (BioVision, Inc., Mountain View, CA, USA); COX-2 Inhibitor Screening Kit (Beyotime Biotech Co., Ltd., China); PVDF (0.45 μ m) (Millipore, Schwalbach, Germany); precolor protein marker (Green BioResearch, LA, USA); fuchsia, Tween 20, acrylamide, and sodium dodecyl sulfate (Solon, OH, USA); protein lysate (RIPA) (Beyotime Biotechnology, Shanghai, China); western blotting membrane regeneration solution (C500031, Sangon Biotech, Shanghai, China); fetal bovine serum (FBS) (GIBCO, NY, USA); RPM1-1640 culture medium (GIBCO, NY, USA); trypsin (Gibco, Grand Island, NY, USA); phorbol myristate acetate (PMA) and penicillin (Sigma-Aldrich, St. Louis, MO); TNF- α ELISA kit (DTA00D) and IL-1 β ELISA kit (DLB50) (R&D Systems, Minnesota, USA); anti-CD86 antibody (ab77276), anti-TNF- α antibody (ab269282), anti-NLRP3 antibody (ab263899), PKA kinase activity assay kit (ab139435), Prostaglandin E2 ELISA Kit (ab133021), and cAMP assay kit (Competitive ELISA) (ab234585) (Abcam, Cambridge, UK); anti-IL-1 β antibody (12703), anti-phos-p65 (Ser536) antibody (3033), anti-p65 antibody (8242), anti-phos-I κ B α (Ser32) antibody (2859), anti-I κ B α antibody (9242), anti-COX-2 antibody (12282) (CST, Boston, USA), and TRIzol (Invitrogen); reverse transcription kit (1708843) and SsoFast EvaGreen Supermix (1725200) (Bio-Rad Laboratories, California, USA); primer (Sangon Biotech Co., Ltd. Shanghai, China); BD Cytofix/Cytoperm™ Fixation/Permeabilization Kit (554714) (BD, New Jersey, USA); ionomycin (SQ23377) (MCE, Shanghai, China); MSU (Invivogen, France); Berthold LB941 microporous plate-type multifunctional enzyme labeling instrument; flow cytometry (Beckman DxFLEX, USA); real-time fluorescence quantitative PCR instrument (ABI 7500, ABI, USA).

Techniques: Western Blot

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Cichoric Acid Ameliorates Monosodium Urate-Induced Inflammatory Response by Reducing NLRP3 Inflammasome Activation via Inhibition of NF- k B Signaling Pathway

doi: 10.1155/2021/8868527

Figure Lengend Snippet: Inhibitory effects of CA on IL-1 β and TNF- α secretion in the THP-M cells. The culture medium was collected, and the levels of IL-1 β (a, c) and TNF- α (b, d) were determined by ELISA. Effect of CA on the mRNA expressions of iNOS and CD86 in THP-M cells. The mRNA levels of iNOS (e) and CD86 (f) were detected by RT-PCR. The results are presented as mean ± SD ( n = 3). ∗ p < 0.05, ∗∗ p < 0.01 vs. the MUS group.

Article Snippet: The specifics of the other materials used in the study are as follows: COX-1 Inhibitor Screening Kit (Fluorometric) (BioVision, Inc., Mountain View, CA, USA); COX-2 Inhibitor Screening Kit (Beyotime Biotech Co., Ltd., China); PVDF (0.45 μ m) (Millipore, Schwalbach, Germany); precolor protein marker (Green BioResearch, LA, USA); fuchsia, Tween 20, acrylamide, and sodium dodecyl sulfate (Solon, OH, USA); protein lysate (RIPA) (Beyotime Biotechnology, Shanghai, China); western blotting membrane regeneration solution (C500031, Sangon Biotech, Shanghai, China); fetal bovine serum (FBS) (GIBCO, NY, USA); RPM1-1640 culture medium (GIBCO, NY, USA); trypsin (Gibco, Grand Island, NY, USA); phorbol myristate acetate (PMA) and penicillin (Sigma-Aldrich, St. Louis, MO); TNF- α ELISA kit (DTA00D) and IL-1 β ELISA kit (DLB50) (R&D Systems, Minnesota, USA); anti-CD86 antibody (ab77276), anti-TNF- α antibody (ab269282), anti-NLRP3 antibody (ab263899), PKA kinase activity assay kit (ab139435), Prostaglandin E2 ELISA Kit (ab133021), and cAMP assay kit (Competitive ELISA) (ab234585) (Abcam, Cambridge, UK); anti-IL-1 β antibody (12703), anti-phos-p65 (Ser536) antibody (3033), anti-p65 antibody (8242), anti-phos-I κ B α (Ser32) antibody (2859), anti-I κ B α antibody (9242), anti-COX-2 antibody (12282) (CST, Boston, USA), and TRIzol (Invitrogen); reverse transcription kit (1708843) and SsoFast EvaGreen Supermix (1725200) (Bio-Rad Laboratories, California, USA); primer (Sangon Biotech Co., Ltd. Shanghai, China); BD Cytofix/Cytoperm™ Fixation/Permeabilization Kit (554714) (BD, New Jersey, USA); ionomycin (SQ23377) (MCE, Shanghai, China); MSU (Invivogen, France); Berthold LB941 microporous plate-type multifunctional enzyme labeling instrument; flow cytometry (Beckman DxFLEX, USA); real-time fluorescence quantitative PCR instrument (ABI 7500, ABI, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Effect of Netrin-1 Anti-Inflammatory Factor on Acute Lung Injury in Sepsis Rats

doi: 10.12659/MSM.917279

Figure Lengend Snippet: Effect of sepsis on the expression of inflammatory factors, netrin-1, and its receptor UNC5B in lung tissues. ( A ) The expression of TNF-α, IL-1β, and IL-6 was detected by Western blot analysis. ** P<0.01 vs. control group. ( B ) The expression of netrin-1 and UNC5B was detected by Western blot analysis. ** P<0.01 vs. control group.

Article Snippet: The membranes were blocked with 5% dry nonfat milk in PBST for 1 h and incubated with primary antibodies against TNF-α (cat no. 3707; Cell Signaling Technology, Inc.; dilution, 1: 1000), IL-1β (cat no. 12703; Cell Signaling Technology, Inc.; dilution, 1: 1,000), IL-6 (ab6672; Abcam, USA; dilution, 1: 1000), netrin-1 (ab126729; Abcam, USA; dilution, 1: 1000), UNC5B (ab104871; Abcam, USA; dilution, 1: 500), and GAPDH (cat no. 5174; Cell Signaling Technology, Inc.; dilution, 1: 1000) overnight at 4°C and then incubated with horseradish peroxidase-conjugated goat anti-rabbit secondary antibody (Santa Cruz Biotechnology, Santa Cruz, CA) for 1 h at 37°C.

Techniques: Expressing, Western Blot

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Effect of Netrin-1 Anti-Inflammatory Factor on Acute Lung Injury in Sepsis Rats

doi: 10.12659/MSM.917279

Figure Lengend Snippet: Netrin-1 alleviated the expression of inflammatory factors and UNC5B in LPS-induced BEAS-2B cells. ( A ) Netrin-1 was decreased in BEAS-2B cells treated with LPS. ** P<0.01 vs. 0 μg/ml group. ( B ) The expression of TNF-α, IL-1β, and IL-6 was detected by Western blot analysis after Netrin-1 transfection. ** P<0.01 and *** P<0.001 vs. control group. ( C ) The expression of UNC5B was detected by Western blot analysis after netrin-1 transfection. * P<0.05 and ** P<0.01 vs. control group.

Article Snippet: The membranes were blocked with 5% dry nonfat milk in PBST for 1 h and incubated with primary antibodies against TNF-α (cat no. 3707; Cell Signaling Technology, Inc.; dilution, 1: 1000), IL-1β (cat no. 12703; Cell Signaling Technology, Inc.; dilution, 1: 1,000), IL-6 (ab6672; Abcam, USA; dilution, 1: 1000), netrin-1 (ab126729; Abcam, USA; dilution, 1: 1000), UNC5B (ab104871; Abcam, USA; dilution, 1: 500), and GAPDH (cat no. 5174; Cell Signaling Technology, Inc.; dilution, 1: 1000) overnight at 4°C and then incubated with horseradish peroxidase-conjugated goat anti-rabbit secondary antibody (Santa Cruz Biotechnology, Santa Cruz, CA) for 1 h at 37°C.

Techniques: Expressing, Western Blot, Transfection

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Sinomenine Regulates Inflammatory Response and Oxidative Stress via Nuclear Factor kappa B (NF-κB) and NF-E2-Related Factor 2 (Nrf2) Signaling Pathways in Ankle Fractures in Children

doi: 10.12659/MSM.910740

Figure Lengend Snippet: TNF-α, IL-1β, and IL-6 expression levels in children with or without fractured ankles. ( A ) The mRNA levels of TNF-α, IL-1β, and IL-6 in children with or without fractured ankle were detected by qRT-PCR; ( B ) The protein levels of TNF-α, IL-1β, and IL-6 in children with or without fractured ankles were detected by ELISA assay. Controls: healthy children; Patients: children with fractured ankle. ** p<0.01 vs. Control.

Article Snippet: Primary antibodies: anti-TNF-α (1: 1000; cat. no. 3707), anti-IL-1β (1: 1000; cat. no. 12703), anti-IL-6 (1: 1000; cat. no. 12153), anti- phosphorylated-p38 (p-p38) (1: 1000; cat. no. 1170), and β-actin (1: 5000; cat no. 4970) were obtained from Cell Signaling Technology, Inc., (Danvers, MA, USA) and anti-phosphorylated-NF-κB (65) (p-NF-κB) was purchased from Abcam (Cambridge, MA).

Techniques: Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Sinomenine Regulates Inflammatory Response and Oxidative Stress via Nuclear Factor kappa B (NF-κB) and NF-E2-Related Factor 2 (Nrf2) Signaling Pathways in Ankle Fractures in Children

doi: 10.12659/MSM.910740

Figure Lengend Snippet: Effect of SIN on inflammatory response. After treatment with 0.25, 0.5, or 1 mM for 24 h, mRNA ( A ) and protein ( B ) levels of TNF-α, IL-1β, and IL-6 were detected by qRT-PCR and Western blot, respectively; Protein levels of p-p38 and p-p65 were detected using Western blotting ( C ) and the data were analyzed ( D ). Controls: cells without any treatment; BK: 1 μg/ml BK treatment group; BK + SIN 0.25: 1 μg/ml BK + 0.25 nM SIN treatment group; BK + SIN 0.5: 1 μg/ml BK + 0.5 nM SIN treatment group; BK + SIN 1: 1 μg/ml BK + 1 nM SIN treatment group;** p<0.01 vs. Control; # p<0.05 vs. BK; ## p<0.01 vs. BK.

Article Snippet: Primary antibodies: anti-TNF-α (1: 1000; cat. no. 3707), anti-IL-1β (1: 1000; cat. no. 12703), anti-IL-6 (1: 1000; cat. no. 12153), anti- phosphorylated-p38 (p-p38) (1: 1000; cat. no. 1170), and β-actin (1: 5000; cat no. 4970) were obtained from Cell Signaling Technology, Inc., (Danvers, MA, USA) and anti-phosphorylated-NF-κB (65) (p-NF-κB) was purchased from Abcam (Cambridge, MA).

Techniques: Quantitative RT-PCR, Western Blot

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Electroacupuncture Synergistically Inhibits Proinflammatory Cytokine Production and Improves Cognitive Function in Rats with Cognitive Impairment due to Hepatic Encephalopathy through p38MAPK/STAT3 and TLR4/NF- κ B Signaling Pathways

doi: 10.1155/2021/7992688

Figure Lengend Snippet: Wistar rats were treated with intraperitoneal injections of 1 mL/kg of 35% CCl 4 twice a week for 9 weeks. When HE models were established, the lactulose group (CCl 4 + Lac) and acupuncture combined with medicine group (CCl 4 + CM) were gavaged with 10 mL/kg body weight. EA was given to the EA (CCl 4 + EA) and the combination (CCl 4 + CM) groups 30 min per day with consecutive 21 days. There are 10 rats in the normal group, and 9 rats in each of no-intervention group (CCl 4 ) and treatment groups including EA group (CCl 4 + EA), lactulose group (CCl 4 + Lac), and EA combined with lactulose (CCl 4 + CM) group. Expressions of TNF- α , IL-6, IL-1 β , iNOS, TLR4, MyD88, NF- κ B, p38MAPK, and STAT3 genes were used to evaluate hepatic inflammation in the mRNA level detected by RT-qPCR.

Article Snippet: TNF- α (60291-1-Ig), IL-1 β (12703), IL-6 (12153), TLR4 (14358), MYD88 (4283), NF- κ B (8242), p38MAPK (6417), (p)-p38MAPK (8690), STAT3 (9139), p-STAT3 (9145), and INOS (SBJ_M0506) were purchased from Cell Signaling Technology.

Techniques: Quantitative RT-PCR