Journal: Journal of Inflammation Research

Article Title: Zuo Gui Wan Promotes Remyelination in Multiple Sclerosis by Attenuating MAPK-Mediated Microglial M1 Polarization, an Integrated Network Pharmacology and Experimental Validation Study

doi: 10.2147/JIR.S572873

Figure Lengend Snippet: ZGW promotes remyelination in CPZ-induced demyelinated mice. ( A ) Representative LFB staining of the corpus callosum. ( B ) TEM image of the corpus callosum ultrathin sections. ( C ) Quantification of the LFB-positive area in the corpus callosum. ( D ) Myelin g-ratio (axon diameter/total fiber diameter) measured from TEM images. ( E ) Representative Western blots for MOG, Olig2, CNPase, and β-actin (loading control). ( F – H ) Quantification of MOG, Olig2, and CNPase normalized to β-actin. *P < 0.05, **P < 0.01, ***P < 0.001 versus CPZ, # P < 0.05, ### P < 0.001 versus NC. Data are presented as mean ± SD. (n =3 per group).

Article Snippet: Primary antibodies were as follows, MOG (Abcam, AB242374 ), CNPase (Santa Cruz, sc-166558), Olig2 (Merck, AB9610), Iba-1 (Hunan Aifang, AFW12391 ), β-actin (Proteintech, 66009-1-Ig), iNOS (Proteintech, 22226-1-AP), CD86 (Proteintech, 26903-1-AP), Arg-1 (Proteintech, 16001-1-AP), CD206 (Proteintech, 18704-1-AP), ERK1/2 (Proteintech, 16443-1-AP), p-ERK1/2 (Proteintech, 28733-1-AP), GAPDH (Servicebio, GB11002-100), p38 (CST, 9212S), p-p38 (CST, 4511S), JNK (CST, 9252S), and p-JNK (CST, 4668S).

Techniques: Staining, Western Blot, Control

Journal: Journal of Advanced Research

Article Title: Neonatal sevoflurane exposures inhibits DHHC5-mediated palmitoylation of TfR1 in oligodendrocytes, leading to hypomyelination and neurological impairments

doi: 10.1016/j.jare.2025.02.009

Figure Lengend Snippet: Inhibition of iron dyshomeostasis and ferroptosis completely reversed hypomyelination and neurological impairment following repeated sevoflurane exposures. (A) Trajectory heat map of the test phase of Barnes maze in P32 mice. (B) Mean latency of the training phase in Barnes maze. (C) Mean latency [F (2, 21) = 8.954, P = 0.0015] and (D) time spent in the target quadrant [F (2, 21) = 6.515, P = 0.0063] during the test phase in Barnes maze. (E) Time on rotarod of P32 mice (Kruskal-Wallis test statistic = 8.163, P = 0.0169). (F) Representative electron micrographs in the CC on P32. (G) Relationship between axon diameter and g -ratio. (H) Quantification of g -ratio [Kruskal-Wallis test statistic = 53.49, P < 0.0001]. (I, J) Representative immunofluorescent staining and quantification for MBP in the CC [F (2, 9) = 8.831, P = 0.0075]. (K, L) Western blotting and quantification analysis of the MBP [F (2, 6) = 40.49, P = 0.0003], CNP [F (2, 6) = 17.87, P = 0.003], and 4-HNE [F (2, 6) = 10.29, P = 0.0115] protein levels in the CC. (M−Q) Representative immunofluorescent staining and quantification of 4-HNE[F (3, 20) = 9.316, P = 0.0005], Liperfluo [F (3, 9) = 6.262, P = 0.0139], TUNEL[F (3, 12) = 53.13, P < 0.0001], MBP[F (3, 17) = 7.546, P = 0.002], and Olig2 in primary cultured OLs. (R) Cell viability in OLs [F (3, 16) = 19.93, P < 0.0001]. Horizontal and error bars represent the mean and SEM, respectively. CC, corpus callosum. MBP, myelin basic protein. CNP, 2′,3′-cyclic-nucleotide 3′-phosphodiesterase. 4-HNE, 4-hydroxynoneal. OLs, oligodendrocytes. For panel B, two-way ANOVA followed by Tukey’s post hoc test was used; for panels E and H, Kruskal-Wallis test with Dunn’s post-hoc test was used; for the remaining panels, one-way ANOVA was used.

Article Snippet: Then 20/50 mg protein was loaded onto denaturing SDS-PAGE gel and subsequently transferred to a 0.22 μm polyvinylidene fluoride (PVDF) membrane, which was blocked in 5 % milk for 2 h and incubated at 4 °C overnight with primary antibodies, including against MBP (1:2,000), 4-HNE (1:1,000), GPX4(1:1000), 2′,3′-cyclic-nucleotide 3′-phosphodiesterase (CNP, 1:1,000, CST, #2986S), DHHC5 (1:1,000, Proteintech, #21324-1-AP), TfR1 (1:1,000, Abcam, #ab84036), Na,K-ATPase (1:1000, CST, #3010S) and GAPDH (1:1,000, HUABIO, #ET1601-4).

Techniques: Inhibition, Staining, Western Blot, TUNEL Assay, Cell Culture

Journal: Journal of Advanced Research

Article Title: Neonatal sevoflurane exposures inhibits DHHC5-mediated palmitoylation of TfR1 in oligodendrocytes, leading to hypomyelination and neurological impairments

doi: 10.1016/j.jare.2025.02.009

Figure Lengend Snippet: DHHC5 over-expression in oligodendrocyte lineage cells rescued sevoflurane-induced cognitive impairment, hypomyelination and dysfunction of oligodendrocytes. (A) Experimental design. (B) Schematic illustration of viral microinjection. (C) Time on rotarod of P32 mice [Kruskal-Wallis test statistic = 10.53, P = 0.0052]. (D) Trajectory heat map of Barnes maze in the test phase. (E) Mean latency during the training phase [F (2, 28) = 6.614, P = 0.0044]. (F) Latency [F (2.000, 12.10) = 13.58, P = 0.0008] and (G) the total time in target zone during test phase. (H-J) Ultrastructure of myelination under transmission electron microscopy and g-ratio measurement [Kruskal-Wallis test statistic = 55.38P < 0.0001]. (K, L) Representative immunofluorescent staining and quantification analysis of CC1 and MBP in the CC, PFC and hippocampus (CA1). (M, N) Western blotting and analysis of MBP [F (2, 6) = 18.96, P = 0.0025], CNP [F (2, 6) = 25.64, P = 0.0011], and 4-HNE [F (2, 6) = 27.46, P = 0.001] in the CC of P32 mice. (O) Intracellular iron level in the CC on P32 following DHHC5 over-expression [F (2, 22) = 20.06, P < 0.0001]. CC, corpus callosum. PFC, prefrontal cortex. CC1, adenomatous polyposis coli. MBP, myelin basic protein. CNP, 2′,3′-cyclic-nucleotide 3′-phosphodiesterase. 4-HNE, 4-hydroxynoneal. DHHC5, zinc finger DHHC-type palmitoyltransferase 5. Horizontal and error bars represent the mean and SEM, respectively. For panels C and J, Kruskal-Wallis test with Dunn’s post-hoc test was used; for panel F, Brown-Forsythe ANOVA test followed by Games-Howell post-hoc test was used; for the remaining panels, one-way ANOVA was used.

Article Snippet: Then 20/50 mg protein was loaded onto denaturing SDS-PAGE gel and subsequently transferred to a 0.22 μm polyvinylidene fluoride (PVDF) membrane, which was blocked in 5 % milk for 2 h and incubated at 4 °C overnight with primary antibodies, including against MBP (1:2,000), 4-HNE (1:1,000), GPX4(1:1000), 2′,3′-cyclic-nucleotide 3′-phosphodiesterase (CNP, 1:1,000, CST, #2986S), DHHC5 (1:1,000, Proteintech, #21324-1-AP), TfR1 (1:1,000, Abcam, #ab84036), Na,K-ATPase (1:1000, CST, #3010S) and GAPDH (1:1,000, HUABIO, #ET1601-4).

Techniques: Over Expression, Microinjection, Transmission Assay, Electron Microscopy, Staining, Western Blot