Journal: Journal of Cellular and Molecular Medicine

Article Title: Zhi‐zi‐chi decoction mitigates depression by enhancing lncRNA Six3os1 expression and promoting histone H3K4 methylation at the BDNF promoter

doi: 10.1111/jcmm.18365

Figure Lengend Snippet: Six3os1 increases BDNF expression by recruiting KMT2A to the BDNF promoter and promoting histone H3K4me3. (A) Six3os1 expression in the hippocampal tissue of control and CUMS mice detected by RT‐qPCR. * p < 0.05 compared with control neurons, # p < 0.05 compared with untreated CORT‐stimulated neurons, (B) Correlation analysis of Six3os1 and BDNF expression, with 12 mice in each group. (C) H3K4me3 modification in the BDNF promoter region analysed by UCSC database. (D) Venn diagram of genes encoding H3K4me3‐associated transferases in the GSEA‐MSigDB database. (E) The binding between KMT2A and Six3os1 predicted by catRAPID website. (F) Six3os1 expression in primary hippocampal neurons treated with oe‐Six3os1 or sh‐Six3os1 detected by RT‐qPCR. * p < 0.05 compared with neurons transduced with oe‐NC, # p < 0.05 compared with neurons transduced with sh‐NC. (G) Western blot of BDNF and H3K4me3 proteins in primary hippocampal neurons treated with sh‐Six3os1.* p < 0.05 compared with neurons transduced with oe‐NC, # p < 0.05 compared with neurons transduced with sh‐NC. (H) RIP assay of Six3os1 binding to KMT2A protein in primary hippocampal neurons. * p < 0.05 compared with IgG. (I) ChIP assay of KMT2A and H3K4me3 enrichment in the BDNF promoter region in primary hippocampal neurons. (J) RNA pull‐down experiment to detect the binding of Six3os1 and KMT2A promoter in primary hippocampal neurons. * p < 0.05 compared with neurons transduced with oe‐NC or Bio‐probe‐NC, # p < 0.05 compared with neurons transduced with sh‐NC. The cell experiment was repeated three times.

Article Snippet: RiboTM lncRNA Six3os1 FISH probe mixture (red) (RiboBio) was used for this assay.

Techniques: Expressing, Control, Quantitative RT-PCR, Modification, Binding Assay, Transduction, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: Zhi‐zi‐chi decoction mitigates depression by enhancing lncRNA Six3os1 expression and promoting histone H3K4 methylation at the BDNF promoter

doi: 10.1111/jcmm.18365

Figure Lengend Snippet: Six3os1 elevates BDNF expression and alleviates CORT‐induced hippocampal neuron injury. (A) BDNF knockdown efficiency in hippocampal neurons detected by RT‐qPCR. * p < 0.05 compared with neurons transduced with sh‐NC. (B) Six3os1 and BDNF expression in CORT‐stimulated hippocampal neurons treated with oe‐Six3os1 or combined with sh‐BDNF detected by RT‐qPCR. (C) Western blot of BDNF protein in CORT‐stimulated hippocampal neurons treated with oe‐Six3os1 or combined with sh‐BDNF. (D) CCK‐8 assay of cell viability of CORT‐stimulated hippocampal neurons treated with oe‐Six3os1 or combined with sh‐BDNF. (E) Apoptosis of CORT‐stimulated hippocampal neurons treated with oe‐Six3os1 or combined with sh‐BDNF measured by flow cytometry. (F) Levels of TNF‐α, IL‐1β and IL‐6 in the supernatant of CORT‐stimulated hippocampal neurons treated with oe‐Six3os1 or combined with sh‐BDNF detected by ELISA. (G) SOD, GSH and CAT activities and MDA production in the supernatant of CORT‐stimulated hippocampal neurons treated with oe‐Six3os1 or combined with sh‐BDNF detected by ELISA. * p < 0.05 compared with CORT‐stimulated hippocampal neurons treated with oe‐NC + sh‐NC, # p < 0.05 compared with neurons transduced with CORT‐stimulated hippocampal neurons treated with oe‐Six3os1 + sh‐NC. The cell experiment was repeated three times.

Article Snippet: RiboTM lncRNA Six3os1 FISH probe mixture (red) (RiboBio) was used for this assay.

Techniques: Expressing, Knockdown, Quantitative RT-PCR, Transduction, Western Blot, CCK-8 Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Zhi‐zi‐chi decoction mitigates depression by enhancing lncRNA Six3os1 expression and promoting histone H3K4 methylation at the BDNF promoter

doi: 10.1111/jcmm.18365

Figure Lengend Snippet: ZZCD represses CORT‐induced hippocampal neuron injury by upregulating the Six3os1/BDNF axis. CORT‐stimulated hippocampal neurons were treated with ZZCD + sh‐NC + oe‐NC, ZZCD + sh‐Six3os1 + oe‐NC or ZZCD + sh‐Six3os1 + oe‐BDNF. (A) Expression of Six3os1 and BDNF in hippocampal neurons detected by RT‐qPCR. (B) Western blot of BDNF and H3K4me3 proteins in hippocampal neurons. (C) Viability of hippocampal neurons detected by CCK‐8 assay. (D) Apoptosis of hippocampal neurons detected by flow cytometry. (E) Levels of TNF‐α, IL‐1β and IL‐6 in the supernatant of hippocampal neurons detected by ELISA. (F) SOD, GSH and CAT activities and MDA production in the supernatant of hippocampal neurons detected by ELISA. * p < 0.05 compared with CORT‐stimulated hippocampal neurons treated with oe‐NC + sh‐NC, # p < 0.05 compared with CORT‐stimulated hippocampal neurons treated with ZZCD + oe‐NC + sh‐NC, & p < 0.05 compared with CORT‐stimulated hippocampal neurons treated with. ZZCD + oe‐Six3os1 + sh‐NC. The cell experiment was repeated three times.

Article Snippet: RiboTM lncRNA Six3os1 FISH probe mixture (red) (RiboBio) was used for this assay.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Zhi‐zi‐chi decoction mitigates depression by enhancing lncRNA Six3os1 expression and promoting histone H3K4 methylation at the BDNF promoter

doi: 10.1111/jcmm.18365

Figure Lengend Snippet: ZZCD alleviates depression‐like behaviours and neuron injury in mice by upregulating the Six3os1/BDNF axis. CUMS mice were treated with ZZCD + sh‐NC + oe‐NC, ZZCD + sh‐Six3os1 + oe‐NC or ZZCD + sh‐Six3os1 + oe‐BDNF. (A) Sucrose preference of CUMS mice tested by SPT. (B) Retention time in the open field centre of CUMS mice tested by OFT. (C) Immobility time of tail suspension of CUMS mice tested by TST. (D) Immobility time of forced swimming of CUMS mice tested by FST. (E) Body weight of CUMS mice. (F) Neuron injury in hippocampal tissues of CUMS mice analysed by HE, Nissl and TUNEL staining. (G) Statistical analysis of pathological scores of hippocampal tissues of CUMS mice. (H) Statistical analysis of Nissl‐positive neurons in hippocampal tissues of CUMS mice. (I) Statistical analysis of TUNEL‐positive neurons in the hippocampal tissue of CUMS mice. (J) Expression of Six3os1 and BDNF in hippocampal tissues of CUMS mice detected by RT‐qPCR. (K) Western blot of BDNF and H3K4me3 proteins in hippocampal tissues of CUMS mice. (L) Levels of TNF‐α, IL‐1β and IL‐6 in mouse serum detected by ELISA. (M) SOD, GSH and CAT activities and MDA production in mouse serum detected by ELISA. n = 12 mice for each treatment. * p < 0.05 compared with control mice treated with oe‐NC + sh‐NC, # p < 0.05 compared with CUMS mice treated with oe‐NC + sh‐NC, & p < 0.05 compared with CUMS mice treated with ZZCD + oe‐NC + sh‐NC, ^ p < 0.05 compared with CUMS mice treated with. ZZCD + oe‐Six3os1 + sh‐NC.

Article Snippet: RiboTM lncRNA Six3os1 FISH probe mixture (red) (RiboBio) was used for this assay.

Techniques: Suspension, TUNEL Assay, Staining, Expressing, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Control