Journal: Molecular Medicine Reports

Article Title: Diospyros lotus leaf extract and its main component myricitrin inhibit itch‑related IL‑6 and IL‑31 by suppressing microglial inflammation and microglial‑mediated astrocyte activation

doi: 10.3892/mmr.2024.13303

Figure Lengend Snippet: Effects of DLE and MC on MAPKs signaling pathway in LPS-stimulated microglia. Microglia were pretreated with DLE or MC at indicated concentrations and stimulated with LPS for 30 min. Relative expression levels of phosphorylated ERK, JNK and p38 with their non-phosphorylated counterparts were evaluated by western blot analysis. Each bar represents the mean ± SD (n=3). # P<0.05 vs. untreated cells; *P<0.05 vs. LPS alone treated cells; ns, no significant difference compared to LPS alone treated cells. DLE, Diospyros lotus leaf extract; MC, myricitrin; LPS, lipopolysaccharide; p-, phosphorylated.

Article Snippet: The following reagents and materials were purchased from the specified suppliers: LPS, Griess reagent and protease inhibitors from MilliporeSigma; myricitrin from Tokyo Chemical Industry; Quanti-MAX WST-8 Cell Viability Assay Kit and WestGlow FEMTO Chemiluminescent substrate from Biomax Ltd.; radio-immunoprecipitation assay (RIPA) buffer, IL-33, inositol 1,4,5-trisphosphate receptor 1 (IP3R1), lipocalin-2 (LCN2), glial fibrillary acidic protein (GFAP), goat anti-rabbit IgG Alexa Fluor 488 antibodies and goat anti-mouse IgG Alexa Fluor 488 from Thermo Fisher Scientific, Inc.; IL-31 antibodies from Abcam; β-actin, phosphorylated (p-)IκBα, IκBα, p-NF-κB, NF-κB, p-JNK, JNK, p-p38, p38, oncostatin M receptor (OSMR), Toll-like receptor 4 (TLR4), IL-6, interleukin 31 receptor a (IL31RA) antibodies and IL-31 short interfering (si)RNA from Santa Cruz Biotechnology, Inc.; and ProLong Gold Antifade Reagent with DAPI, p-Ikk, Ikk, p-JAK1, JAK1 p-STAT3, STAT3, p-ERK and ERK antibodies from Cell Signaling Technology, Inc.

Techniques: Expressing, Western Blot

Journal: Molecular Medicine Reports

Article Title: Diospyros lotus leaf extract and its main component myricitrin inhibit itch‑related IL‑6 and IL‑31 by suppressing microglial inflammation and microglial‑mediated astrocyte activation

doi: 10.3892/mmr.2024.13303

Figure Lengend Snippet: Schematic diagram of the mechanism involved in the anti-itch effect of DLE and MC on astrocytes and microglia. DLE, Diospyros lotus leaf extract; MC, myricitrin; gp130, glycoprotein 130; TLR4, toll-like receptor 4; JAK1, Janus kinase; p38, p38 mitogen-activated protein kinases; LSMCM, LPS-stimulated microglia-conditioned media; IP3R1, inositol 1,4,5-trisphosphate receptor 1, TRPC, transient receptor potential canonical; IL-31RA, interleukin 31 receptor a; OSMR, oncostatin M receptor; LCN2, lipocalin-2; GFAP, glial fibrillary acidic protein.

Article Snippet: The following reagents and materials were purchased from the specified suppliers: LPS, Griess reagent and protease inhibitors from MilliporeSigma; myricitrin from Tokyo Chemical Industry; Quanti-MAX WST-8 Cell Viability Assay Kit and WestGlow FEMTO Chemiluminescent substrate from Biomax Ltd.; radio-immunoprecipitation assay (RIPA) buffer, IL-33, inositol 1,4,5-trisphosphate receptor 1 (IP3R1), lipocalin-2 (LCN2), glial fibrillary acidic protein (GFAP), goat anti-rabbit IgG Alexa Fluor 488 antibodies and goat anti-mouse IgG Alexa Fluor 488 from Thermo Fisher Scientific, Inc.; IL-31 antibodies from Abcam; β-actin, phosphorylated (p-)IκBα, IκBα, p-NF-κB, NF-κB, p-JNK, JNK, p-p38, p38, oncostatin M receptor (OSMR), Toll-like receptor 4 (TLR4), IL-6, interleukin 31 receptor a (IL31RA) antibodies and IL-31 short interfering (si)RNA from Santa Cruz Biotechnology, Inc.; and ProLong Gold Antifade Reagent with DAPI, p-Ikk, Ikk, p-JAK1, JAK1 p-STAT3, STAT3, p-ERK and ERK antibodies from Cell Signaling Technology, Inc.

Techniques:

Journal: Journal of Nanobiotechnology

Article Title: Umbilical cord blood-derived exosomes attenuate dopaminergic neuron damage of Parkinson's disease mouse model

doi: 10.1186/s12951-024-02773-1

Figure Lengend Snippet: UCB-Exos inhibited hyperphosphorylation of the MAPK p38 and ERK1/2 signaling pathway in vitro. A TH, α-Syn, p-α-Syn, ERK, p-ERK, p38/MAPK, and p-p38/MAPK protein levels in MN9D cells treated with UCB-Exos or the vehicle for 24 h were determined by western blot analysis. Full-length blots are presented in Supplementary Fig. 9. B – F Quantitative results of ( A ). n = 4 for each group, data are presented as mean ± SD, * P < 0.05, ** P < 0.01, *** P < 0.005

Article Snippet: The following primary antibodies were used in this study: anti-HSP70 (10995-1-AP), anti-CD63 (67605-1-Ig), anti-Cyt-c (10993-1-AP), anti-TH (25859-1-AP), anti-α-Syn (10842-1-AP), anti-p-ERK (28733-1-AP), anti-p38/MAPK (14064-1-AP), anti-p-p38/MAPK (28796-1-AP) were purchased from Proteintech (Wuhan, China); anti-p-α-Syn (YP0258) was from ImmunoWay Biotechnology (USA); anti-ERK (abs130092) was from Absin (Shanghai, China); anti-tubulin (AF5012), tubulin-tracker staining solution (C1051S), senescence β-galactosidase staining kit (C0602), and Hoechst 33342 (C1026) was from Beyotime Biotechnology (Shanghai, China).

Techniques: In Vitro, Western Blot

Journal: Journal of Nanobiotechnology

Article Title: Umbilical cord blood-derived exosomes attenuate dopaminergic neuron damage of Parkinson's disease mouse model

doi: 10.1186/s12951-024-02773-1

Figure Lengend Snippet: UCB-Exos inhibited hyperphosphorylation of the MAPK p38 and ERK1/2 signaling pathway in vivo. A TH, α-Syn, p-α-Syn, ERK, p-ERK, p38/MAPK, p-p38/MAPK protein levels in mouse midbrain tissue by western blot analysis. Full-length blots are presented in Supplementary Fig. 10. B – F Quantitative results of ( A ). n = 2 for each group, data are presented as mean ± SD, * P < 0.05, ** P < 0.01, *** P < 0.005. mRNA expression levels of Hspb1 and Ppef2 in mouse midbrain tissue ( G ), MN9D cells ( H ), and SH-SY5Y cells ( I ) were detected by Q-PCR analysis

Article Snippet: The following primary antibodies were used in this study: anti-HSP70 (10995-1-AP), anti-CD63 (67605-1-Ig), anti-Cyt-c (10993-1-AP), anti-TH (25859-1-AP), anti-α-Syn (10842-1-AP), anti-p-ERK (28733-1-AP), anti-p38/MAPK (14064-1-AP), anti-p-p38/MAPK (28796-1-AP) were purchased from Proteintech (Wuhan, China); anti-p-α-Syn (YP0258) was from ImmunoWay Biotechnology (USA); anti-ERK (abs130092) was from Absin (Shanghai, China); anti-tubulin (AF5012), tubulin-tracker staining solution (C1051S), senescence β-galactosidase staining kit (C0602), and Hoechst 33342 (C1026) was from Beyotime Biotechnology (Shanghai, China).

Techniques: In Vivo, Western Blot, Expressing

Journal: Journal of Nanobiotechnology

Article Title: Umbilical cord blood-derived exosomes attenuate dopaminergic neuron damage of Parkinson's disease mouse model

doi: 10.1186/s12951-024-02773-1

Figure Lengend Snippet: The schematic illustration demonstrates the potential neuroprotective effects of UCB-Exos on dopaminergic neurons. The effects are primarily manifested through the significant inhibition of neurotoxin-induced cellular stress responses and the subsequent hyperphosphorylation of MAPK p38 and ERK1/2 signaling pathways. UCB-Exos may modulate the expression of various genes, including Hspb1 , Ppef2, Twist1 , and Drd2 , via the cargo they carry, such as miRNA, mRNA, and proteins. This modulation could potentially inhibit neuronal senescence signaling and safeguard synaptic functions, thereby mitigating Parkinson's Disease (PD)-related motor dysfunction and pathological progression. Image created with Figdraw (Home for Researchers)

Article Snippet: The following primary antibodies were used in this study: anti-HSP70 (10995-1-AP), anti-CD63 (67605-1-Ig), anti-Cyt-c (10993-1-AP), anti-TH (25859-1-AP), anti-α-Syn (10842-1-AP), anti-p-ERK (28733-1-AP), anti-p38/MAPK (14064-1-AP), anti-p-p38/MAPK (28796-1-AP) were purchased from Proteintech (Wuhan, China); anti-p-α-Syn (YP0258) was from ImmunoWay Biotechnology (USA); anti-ERK (abs130092) was from Absin (Shanghai, China); anti-tubulin (AF5012), tubulin-tracker staining solution (C1051S), senescence β-galactosidase staining kit (C0602), and Hoechst 33342 (C1026) was from Beyotime Biotechnology (Shanghai, China).

Techniques: Inhibition, Expressing

Journal: Journal of Nanobiotechnology

Article Title: ACYP2 functions as an innovative nano-therapeutic target to impede the progression of hepatocellular carcinoma by inhibiting the activity of TERT and the KCNN4/ERK pathway

doi: 10.1186/s12951-024-02827-4

Figure Lengend Snippet: Cytoplasmic ACYP2 inhibits HCC progression through inactivating the KCNN4/ERK pathway. A KCNN4 expression was detected by qRT-PCR and correlation analysis its expression with ACYP2 in HCC; B the interaction between ACYP2 and KCNN4 was verified by Co-immunoprecipitation (IP) assay; C the mRNA expression of KCNN4 in si-ACYP2 (si-ACYP2-1) and pcDNA-ACYP2 treated HCC cells was detected by qRT-PCR; D the protein expression of KCNN4 in si-ACYP2 and pcDNA-ACYP2 treated HCC cells was detected by western blotting; E the proliferation abilities of HCC cells in si-NC, si-KCNN4 and si-ACYP2 + si-KCNN4 treatment groups were analyzed by CCK-8; F the apoptotic rates of HCC cells in si-NC, si-KCNN4 and si-ACYP2 + si-KCNN4 treatment groups were detected by flow cytometry; G the cell invasion abilities of si-NC, si-KCNN4 and si-ACYP2 + si-KCNN4 treatment groups were conducted by transwell assay; H the concentration of K + was detected by ion chromatography; I the fluorescence intensity of K + in HCC cells was measured by confocal fluorescence microscopy; J The expression of ERK, p-ERK, p38, p-p38, JNK and p-JNK proteins of si-NC, si-KCNN4 and si-ACYP2 + si-KCNN4 treatment groups were verified respectively by western blotting. Scale bar: 100 μm; * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: GAPDH (Absin, China), ERK and p-ERK (Proteintech, China), P38, p-P38, JNK and p-JNK (Abmart, China), ACYP2, KCNN4, and TERT (Abcam, UK) were the primary antibodies employed in this investigation.

Techniques: Expressing, Quantitative RT-PCR, Immunoprecipitation, Western Blot, CCK-8 Assay, Flow Cytometry, Transwell Assay, Concentration Assay, Ion Chromatography, Fluorescence, Microscopy

Journal: Journal of Nanobiotechnology

Article Title: ACYP2 functions as an innovative nano-therapeutic target to impede the progression of hepatocellular carcinoma by inhibiting the activity of TERT and the KCNN4/ERK pathway

doi: 10.1186/s12951-024-02827-4

Figure Lengend Snippet: Cytoplasmic ACYP2 inhibits HCC progression through inactivating the KCNN4/ERK pathway. A KCNN4 expression was detected by qRT-PCR and correlation analysis its expression with ACYP2 in HCC; B the interaction between ACYP2 and KCNN4 was verified by Co-immunoprecipitation (IP) assay; C the mRNA expression of KCNN4 in si-ACYP2 (si-ACYP2-1) and pcDNA-ACYP2 treated HCC cells was detected by qRT-PCR; D the protein expression of KCNN4 in si-ACYP2 and pcDNA-ACYP2 treated HCC cells was detected by western blotting; E the proliferation abilities of HCC cells in si-NC, si-KCNN4 and si-ACYP2 + si-KCNN4 treatment groups were analyzed by CCK-8; F the apoptotic rates of HCC cells in si-NC, si-KCNN4 and si-ACYP2 + si-KCNN4 treatment groups were detected by flow cytometry; G the cell invasion abilities of si-NC, si-KCNN4 and si-ACYP2 + si-KCNN4 treatment groups were conducted by transwell assay; H the concentration of K + was detected by ion chromatography; I the fluorescence intensity of K + in HCC cells was measured by confocal fluorescence microscopy; J The expression of ERK, p-ERK, p38, p-p38, JNK and p-JNK proteins of si-NC, si-KCNN4 and si-ACYP2 + si-KCNN4 treatment groups were verified respectively by western blotting. Scale bar: 100 μm; * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: GAPDH (Absin, China), ERK and p-ERK (Proteintech, China), P38, p-P38, JNK and p-JNK (Abmart, China), ACYP2, KCNN4, and TERT (Abcam, UK) were the primary antibodies employed in this investigation.

Techniques: Expressing, Quantitative RT-PCR, Immunoprecipitation, Western Blot, CCK-8 Assay, Flow Cytometry, Transwell Assay, Concentration Assay, Ion Chromatography, Fluorescence, Microscopy