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