Journal: Experimental and therapeutic medicine

Article Title: Luteolin suppresses TNF-α-induced inflammatory injury and senescence of nucleus pulposus cells via the Sirt6/NF-κB pathway.

doi: 10.3892/etm.2022.11396

Figure Lengend Snippet: Figure 1. Luteolin enhances the viability of TNF‑α‑induced HNPCs. (A) Chemical structure of luteolin. (B) Effect of different concentrations (1, 2 and 4 µM) of luteolin on HNPC viability. (C) Effect of luteolin on TNF‑α‑suppressed HNPC viability. ***P<0.001 vs. control. ##P<0.01 vs. TNF‑α. TNF‑α, tumor necrosis factor‑α; HNPCs, human nucleus pulposus cells.

Article Snippet: Immortalized human nucleus pulposus cells (HNPCs) were obtained from AcceGen Biotechnology (cat. no. ABI‐TC102D).

Techniques: Control

Journal: Experimental and therapeutic medicine

Article Title: Luteolin suppresses TNF-α-induced inflammatory injury and senescence of nucleus pulposus cells via the Sirt6/NF-κB pathway.

doi: 10.3892/etm.2022.11396

Figure Lengend Snippet: Figure 2. Luteolin inhibits TNF‑α‑induced HNPC inflammatory injury. (A and B) Expression levels of inflammatory cytokines IL‑1β (A) and IL‑6 (B) were detected by ELISA. (C) TUNEL staining was used to detect the effect of luteolin on TNF‑α‑induced apoptosis. (D) Expression levels of Bcl‑2, Bax and cleaved caspase‑3 protein were detected by western blot analysis. ***P<0.001 vs. control. #P<0.05, ##P<0.01 and ###P<0.001 vs. TNF‑α. TNF‑α, tumor necrosis factor‑α; HNPCs, human nucleus pulposus cells.

Article Snippet: Immortalized human nucleus pulposus cells (HNPCs) were obtained from AcceGen Biotechnology (cat. no. ABI‐TC102D).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, TUNEL Assay, Staining, Western Blot, Control

Journal: Experimental and therapeutic medicine

Article Title: Luteolin suppresses TNF-α-induced inflammatory injury and senescence of nucleus pulposus cells via the Sirt6/NF-κB pathway.

doi: 10.3892/etm.2022.11396

Figure Lengend Snippet: Figure 3. Luteolin suppresses TNF‑α‑induced senescence of HNPCs. (A) Senescence β‑galactosidase staining kit was used to detect the activity level of β‑galactosidase in HNPCs. (B) ELISA kit was performed to detect the activity of telomerase. (C) The expression levels of senescence related proteins (p16 and p21) were examined via western blot analysis. (D) Expression of p53 was detected by western blot analysis. ***P<0.001 vs. control. #P<0.05, ##P<0.01 and ###P<0.001 vs. TNF‑α. TNF‑α, tumor necrosis factor‑α; HNPCs, human nucleus pulposus cells.

Article Snippet: Immortalized human nucleus pulposus cells (HNPCs) were obtained from AcceGen Biotechnology (cat. no. ABI‐TC102D).

Techniques: Staining, Activity Assay, Enzyme-linked Immunosorbent Assay, Expressing, Western Blot, Control

Journal: Experimental and therapeutic medicine

Article Title: Luteolin suppresses TNF-α-induced inflammatory injury and senescence of nucleus pulposus cells via the Sirt6/NF-κB pathway.

doi: 10.3892/etm.2022.11396

Figure Lengend Snippet: Figure 4. Luteolin regulates the Sirt6/NF‑κB pathway. (A) The expression levels of Sirt6/NF‑κB pathway‑related proteins (Sirt6, p‑NF‑κB and p‑NF‑κB p65) were examined by western blot analysis. (B) Western blot analysis was used to examine histone acetylation‑related H3K9ac expression level. (C) Effect of luteolin on TNF‑α‑induced Sirt6 activity was detected by SIRT6 activity assay kit. (D) Interaction between Sirt6 and luteolin was predicted by molecular docking. ***P<0.001 vs. control. #P<0.05, ##P<0.01 and ###P<0.001 vs. TNF‑α. TNF‑α, tumor necrosis factor‑α; HNPCs, human nucleus pulposus cells.

Article Snippet: Immortalized human nucleus pulposus cells (HNPCs) were obtained from AcceGen Biotechnology (cat. no. ABI‐TC102D).

Techniques: Expressing, Western Blot, Activity Assay, Control

Journal: Experimental and therapeutic medicine

Article Title: Luteolin suppresses TNF-α-induced inflammatory injury and senescence of nucleus pulposus cells via the Sirt6/NF-κB pathway.

doi: 10.3892/etm.2022.11396

Figure Lengend Snippet: Figure 5. Sirt6 knockdown partially reverses the inhibitory effect of luteolin on TNF‑α‑induced inflammatory damage of HNPCs. (A and B) The Sirt6 protein (A) and mRNA (B) expression levels were detected by western blot analysis and RT‑qPCR, respectively. (C‑E) Effects of Sirt6 knockdown on cell viability (C) and intracellular IL‑1β (D) and IL‑6 (E) expression levels. (F) TUNEL staining was used to detect the effect of Sirt6 knockdown on HNPC apoptosis. (G) Expression levels of Bcl‑2, Bax and cleaved caspase 3 protein were detected by western blot analysis. ***P<0.001 vs. control or si‑NC. ##P<0.01 and ###P<0.001 vs. TNF‑α. +P<0.05 ++P<0.01 and +++P<0.001 vs. TNF‑α + luteolin. @@P<0.01 and @@@P<0.001 vs. TNF‑α + luteolin + si‑NC. TNF‑α, tumor necrosis factor‑α; HNPCs, human nucleus pulposus cells; Sirt6, sirtuin 6.

Article Snippet: Immortalized human nucleus pulposus cells (HNPCs) were obtained from AcceGen Biotechnology (cat. no. ABI‐TC102D).

Techniques: Knockdown, Expressing, Western Blot, TUNEL Assay, Staining, Control

Journal: Experimental and therapeutic medicine

Article Title: Luteolin suppresses TNF-α-induced inflammatory injury and senescence of nucleus pulposus cells via the Sirt6/NF-κB pathway.

doi: 10.3892/etm.2022.11396

Figure Lengend Snippet: Figure 6. Knockdown of Sirt6 partially reverses the inhibitory effect of luteolin on TNF‑α‑induced senescence of HNPCs. (A) The activity level of β‑galactosidase in HNPCs was assayed by senescence β‑galactosidase staining. (B) ELISA kit was performed to detect the activity of telomerase. (C) Expression levels of senescence‑related proteins (p16 and p21) were examined via western blot analysis. (D) Expression of p53 was detected by western blot analysis. ***P<0.001 vs. control. ###P<0.001 vs. TNF‑α. +++P<0.001 vs. TNF‑α + luteolin. @@P<0.01, @@@P<0.001 vs. TNF‑α + luteolin + si‑NC. TNF‑α, tumor necrosis factor‑α; HNPCs, human nucleus pulposus cells; Sirt6, sirtuin 6.

Article Snippet: Immortalized human nucleus pulposus cells (HNPCs) were obtained from AcceGen Biotechnology (cat. no. ABI‐TC102D).

Techniques: Knockdown, Activity Assay, Staining, Enzyme-linked Immunosorbent Assay, Expressing, Western Blot, Control

Journal: International Journal of Molecular Medicine

Article Title: Knockdown of miR-222 inhibits inflammation and the apoptosis of LPS-stimulated human intervertebral disc nucleus pulposus cells

doi: 10.3892/ijmm.2019.4314

Figure Lengend Snippet: LPS-induced nucleus pulposus cell apoptosis is inhibited by miR-222 inhibitor. (A) miR-222 level in human intervertebral disc tissues was detected by reverse transcription-quantitative polymerase chain reaction. (B) miR-222 level in lipopolysaccharide-stimulated nucleus pulposus cells was detected using reverse transcription-quantitative polymerase chain reaction. (C) Transfection efficiency of miR-222 mock, mimics and inhibitor was determined by reverse transcription-quantitative polymerase chain reaction. (D) Nucleus pulposus cell apoptosis was analyzed by flow cytometry. * P<0.05 and ** P<0.01 vs. control; # P<0.05 and ## P<0.01 vs. cells stimulated with LPS only. LPS, lipopolysaccharide.

Article Snippet: Human nucleus pulposus (NP, cat. no. 4800) cells were obtained from ScienCell Research Laboratories and cultured in Dulbecco's modified Eagle's medium (DMEM, Gibco; Thermo Fisher Scientific) containing 10% fetal bovine serum (FBS, Gibco; Thermo Fisher Scientific) and antibiotics (1% penicillin/streptomycin, BBI Life Sciences) at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: Reverse Transcription, Real-time Polymerase Chain Reaction, Transfection, Flow Cytometry, Control

Journal: International Journal of Molecular Medicine

Article Title: Knockdown of miR-222 inhibits inflammation and the apoptosis of LPS-stimulated human intervertebral disc nucleus pulposus cells

doi: 10.3892/ijmm.2019.4314

Figure Lengend Snippet: Effects of miR-222 on TNF-α, IL-1β and IL-6, collagen II and aggrecan expression levels in LPS-treated nucleus pulposus cells. (A) TNF-α, (B) IL-1β and (C) IL-6 expression levels in nucleus pulposus cells treated with LPS were detected by ELISA. The expression levels of (D) collagen II and (E) aggrecan in nucleus pulposus cells treated with LPS were detected using reverse transcription-quantitative polymerase chain reaction * P<0.05 and ** P<0.01 vs. control, # P<0.05 and ## P<0.01 vs. cells stimulated with LPS only. TNF-α, tumor necrosis factor- α; IL, interleukin; LPS, lipopolysaccharide.

Article Snippet: Human nucleus pulposus (NP, cat. no. 4800) cells were obtained from ScienCell Research Laboratories and cultured in Dulbecco's modified Eagle's medium (DMEM, Gibco; Thermo Fisher Scientific) containing 10% fetal bovine serum (FBS, Gibco; Thermo Fisher Scientific) and antibiotics (1% penicillin/streptomycin, BBI Life Sciences) at 37°C in a humidified atmosphere of 5% CO 2 .

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

Journal: International Journal of Molecular Medicine

Article Title: Knockdown of miR-222 inhibits inflammation and the apoptosis of LPS-stimulated human intervertebral disc nucleus pulposus cells

doi: 10.3892/ijmm.2019.4314

Figure Lengend Snippet: Overexpression of TIMP3 reverses the apoptosis of LPS-stimulated nucleus pulposus cells induced by miR-222 inhibitor. (A) TIMP3 mRNA levels was determined by reverse transcription-quantitative polymerase chain reaction. (B) TIMP3 protein levels were determined by western blot analysis. (C) The relative TIMP3 protein level is presented. (D) Cell apoptosis was analyzed by flow cytometry. ** P<0.01 vs. control; ## P<0.01 vs. cells stimulated with LPS only; && P<0.01 vs. inhibitor + siNC.

Article Snippet: Human nucleus pulposus (NP, cat. no. 4800) cells were obtained from ScienCell Research Laboratories and cultured in Dulbecco's modified Eagle's medium (DMEM, Gibco; Thermo Fisher Scientific) containing 10% fetal bovine serum (FBS, Gibco; Thermo Fisher Scientific) and antibiotics (1% penicillin/streptomycin, BBI Life Sciences) at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: Over Expression, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Flow Cytometry, Control

Journal: Molecular Medicine Reports

Article Title: Upregulation of SIRT1 by Evodiamine activates PI3K/AKT pathway and blocks intervertebral disc degeneration

doi: 10.3892/mmr.2022.12781

Figure Lengend Snippet: Evo inhibits LPS-induced NPCs apoptosis. (A) Cell viability of NPCs incubated with Evo at 5, 10, 20 and 40 µM for 24 h. (B) CCK-8 assay and (C and D) TUNEL staining were used to detect the effects of Evo on the viability and apoptosis of LPS-induced NPCs. (E) Caspase-3 activity was detected in NPCs and (F) the expression levels of apoptosis-related proteins (Bax and Bcl-2) were detected by western blotting. **P<0.01 and ***P<0.001 vs. Control. ## P<0.01 and ### P<0.001 vs. LPS. Evo, evodiamine; LPS, lipopolysaccharide; NPCs, human nucleus pulposus cells.

Article Snippet: Immortalized human nucleus pulposus cells (NPCs; iCell Bioscience Inc.; cat. no. iCell-0028a) were cultured in DMEM (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.), 100 U/ml penicillin and 100 µg/ml streptomycin (Invitrogen; Thermo Fisher Scientific, Inc.) at 37°C in 5% CO 2 .

Techniques: Incubation, CCK-8 Assay, TUNEL Assay, Staining, Activity Assay, Expressing, Western Blot

Journal: Molecular Medicine Reports

Article Title: Upregulation of SIRT1 by Evodiamine activates PI3K/AKT pathway and blocks intervertebral disc degeneration

doi: 10.3892/mmr.2022.12781

Figure Lengend Snippet: Evo effectively alleviated LPS-induced extracellular matrix degradation and inflammation in NPCs. (A) Western blotting was applied to examine the expression of MMP-13, Aggrecan, collagen II and SOX-9 proteins. The concentrations of (B) TNF-α and (C) IL-6 in NPCs were determined by ELISA. ***P<0.001 vs. Control. # P<0.05, ## P<0.01 and ### P<0.001 vs. LPS. Evo, evodiamine; LPS, lipopolysaccharide; NPCs, human nucleus pulposus cells; SOX-9, sry-type high-mobility-group box 9; MMP, matrix metalloproteinase; TNF, tumor necrosis factor; IL, interleukin.

Article Snippet: Immortalized human nucleus pulposus cells (NPCs; iCell Bioscience Inc.; cat. no. iCell-0028a) were cultured in DMEM (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.), 100 U/ml penicillin and 100 µg/ml streptomycin (Invitrogen; Thermo Fisher Scientific, Inc.) at 37°C in 5% CO 2 .

Techniques: Western Blot, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Molecular Medicine Reports

Article Title: Upregulation of SIRT1 by Evodiamine activates PI3K/AKT pathway and blocks intervertebral disc degeneration

doi: 10.3892/mmr.2022.12781

Figure Lengend Snippet: Evo activates the PI3K/AKT pathway by upregulating SIRT1. (A) Western blotting was used to detect the expression level of SIRT1 protein and (B) the effect of Evo on the P13K/AKT pathway-related proteins (P13K, AKT, p-P13K and p-AKT) in LPS-induced NPCs. ***P<0.001 vs. Control. ## P<0.01 and ### P<0.001 vs. LPS. Evo, evodiamine; SIRT1, Sirtuin 1; p-, phosphorylated; LPS, lipopolysaccharide; NPCs, human nucleus pulposus cells.

Article Snippet: Immortalized human nucleus pulposus cells (NPCs; iCell Bioscience Inc.; cat. no. iCell-0028a) were cultured in DMEM (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.), 100 U/ml penicillin and 100 µg/ml streptomycin (Invitrogen; Thermo Fisher Scientific, Inc.) at 37°C in 5% CO 2 .

Techniques: Western Blot, Expressing

Journal: Molecular Medicine Reports

Article Title: Upregulation of SIRT1 by Evodiamine activates PI3K/AKT pathway and blocks intervertebral disc degeneration

doi: 10.3892/mmr.2022.12781

Figure Lengend Snippet: SIRT1 knockdown attenuates the inhibitory effects of Evo on LPS-induced apoptosis in NPCs. Western blotting was performed to detect the interference efficiency of (A) SIRT1 and (B) the expression levels of P13K, AKT, p-P13K and p-AKT protein. (C) CCK-8 assay and (D and E) TUNEL staining were conducted to test the viability and apoptosis level of NPCs and (F) Caspase-3 detection kit was used to detect the activity of caspase-3 in NPCs. (G) The expression level of Bax and Bcl-2 protein in NPCs. ***P<0.001 vs. Control. ### P<0.001 vs. LPS. & P<0.05, && P<0.01 and &&& P<0.001 vs. si-NC. SIRT1, Sirtuin 1; Evo, evodiamine; LPS, lipopolysaccharide; NPCs, human nucleus pulposus cells; p-, phosphorylated.

Article Snippet: Immortalized human nucleus pulposus cells (NPCs; iCell Bioscience Inc.; cat. no. iCell-0028a) were cultured in DMEM (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.), 100 U/ml penicillin and 100 µg/ml streptomycin (Invitrogen; Thermo Fisher Scientific, Inc.) at 37°C in 5% CO 2 .

Techniques: Western Blot, Expressing, CCK-8 Assay, TUNEL Assay, Staining, Activity Assay

Journal: Molecular Medicine Reports

Article Title: Upregulation of SIRT1 by Evodiamine activates PI3K/AKT pathway and blocks intervertebral disc degeneration

doi: 10.3892/mmr.2022.12781

Figure Lengend Snippet: SIRT1 knockdown attenuates the inhibitory effects of Evo on LPS-induced inflammation and ECM degradation in NPCs. (A) Western blotting was applied to examine the expression of MMP-13, Aggrecan, collagen II and SOX-9 proteins. The concentrations of (B) TNF-α and (C) IL-6 in NPCs were determined by ELISA.***P<0.001 vs. Control. ### P<0.001 vs. LPS. & P<0.05, && P<0.01 and &&& P<0.001 vs. si-NC. SIRT1, Sirtuin 1; Evo, evodiamine; LPS, lipopolysaccharide; ECM, extracellular matrix; NPCs, human nucleus pulposus cells; SOX-9, sry-type high-mobility-group box 9; MMP, matrix metalloproteinase; TNF, tumor necrosis factor; IL, interleukin; si, short interfering; NC, negative control.

Article Snippet: Immortalized human nucleus pulposus cells (NPCs; iCell Bioscience Inc.; cat. no. iCell-0028a) were cultured in DMEM (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.), 100 U/ml penicillin and 100 µg/ml streptomycin (Invitrogen; Thermo Fisher Scientific, Inc.) at 37°C in 5% CO 2 .

Techniques: Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Negative Control

Journal: Materials Today Bio

Article Title: Sequential delivery of sinigrin and dabigatran by an in situ self-stabilizing dynamic hydrogel attenuates intervertebral disc degeneration

doi: 10.1016/j.mtbio.2026.102827

Figure Lengend Snippet: Metabolic regulation of nucleus pulposus vells by SIN and DAB through MAPK and NF-κB pathways. (A) Volcano plot analysis of DE genes (TNF-α vs. TNF-α+SIN). (B) KEGG enrichment analysis of DE genes (TNF-α vs. TNF-α+SIN). (C) GSEA indicated MAPK signalling pathway enrichment. (D) Heatmap of ten selected differentially expressed genes (TNF-α vs. TNF-α+SIN). (E) Western blot analysis of key proteins in the MAPK signalling pathway (p-ERK, p-JNK, p-p38, ERK, JNK, and p38) in the different treatment groups. (F) Quantitative analysis of the Western blot data for MAPK signalling pathway proteins. (G–H) Western blot and quantitative analysis of p-ERK, p-JNK, p-p38, ERK, JNK, and p38 treated with or without TNF-α in the absence or presence of SIN (10 μM) (PCDNA 3.1 vs. TNFR1-OE).(I–L) Integrated analysis of TNF-α vs. TNF-α+DAB DEGs: Volcano plot of DE genes (significance/fold change), KEGG pathway enrichment of DE genes, NF-κB signalling enrichment in GSEA, and heatmap of ten selected differentially expressed genes. (M) Western blot analysis of key proteins in the NF-κB signalling pathway (p-IκBα, p-p65, IκBα, and p65) in the different treatment groups. (N) Quantitative analysis of the Western blot data for the NF-κB signalling pathway proteins. (O–P) Western blot and quantitative analysis of p-p65 and p65 (PCDNA 3.1 vs. TNFR1-OE). (Q) Matched diagram of b and y ions. (R) Western blot analysis of RELA in the different treatment groups. (S) Western blot analysis confirmed the stabilization of p65 by DAB. (T) The affinity between DAB and the p65 target protein was estimated after incubation at different temperatures via Western blot analysis. (n = 3; ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001).

Article Snippet: Immortalized human nucleus pulposus cells (Procell, Cat. No. CP-H097Y) were co-transfected with an NF-κB-responsive firefly luciferase reporter plasmid and a Renilla luciferase internal control plasmid for 8 h. Following transfection, cells were treated overnight with the test compounds at a concentration of 10 μM.

Techniques: Western Blot, Incubation