Journal: CNS Neuroscience & Therapeutics

Article Title: Jolkinolide B Mitigates Cerebral Ischemia–Reperfusion Injury by Promoting Microglial M1 / M2 Polarization Through the JAK2 / STAT3 Signaling Pathway

doi: 10.1111/cns.70653

Figure Lengend Snippet: JB alleviates neurological dysfunction and pathological damage induced by CIRI in MCAO/R rats. (A) Schematic diagram of the experimental workflow. Rats in each group received intraperitoneal injections of JB once daily for 3 consecutive days starting from the onset of reperfusion. Neurological behavior tests were performed 1 h after the final administration. Subsequently, the animals were sacrificed, and brain tissues were rapidly harvested for further analysis. (B, C) Neurological function deficits were assessed using the Longa scoring system and the corner test. n = 18. (D, E) TTC staining was used to evaluate the extent of cerebral infarction and measure the infarct volume in each group. (F–H) HE staining was performed to observe pathological changes in cortical tissue structures across groups. Nissl staining was used to quantify the number of Nissl bodies. Scale bar 50 μm. (I, J) TUNEL staining was conducted to detect neuronal apoptosis in the cerebral cortex. Scale bar 50 μm. n = 6. *** p < 0.001 vs. sham; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. MCAO/R.

Article Snippet: The apoptosis of neuronal cells in rat brain tissue was assessed following the instructions of the TUNEL cell apoptosis detection kit (A113‐01, Vazyme).

Techniques: Staining, TUNEL Assay

Journal: CNS Neuroscience & Therapeutics

Article Title: Jolkinolide B Mitigates Cerebral Ischemia–Reperfusion Injury by Promoting Microglial M1 / M2 Polarization Through the JAK2 / STAT3 Signaling Pathway

doi: 10.1111/cns.70653

Figure Lengend Snippet: JB improved HAPI cell survival and decreased apoptosis under OGD/R conditions. (A) CCK‐8 assay was used to assess JB (0, 2.5, 5, 10, 20, 40 μM) effects on HAPI cell viability. (B) CCK‐8 assay determined the effects of JB (0, 2.5, 5, 10, 20, 40 μM) on the viability of OGD/R‐treated HAPI cells. (C, D) TUNEL assay to determine the effects of JB at concentrations of 2.5, 5, and 10 μM on apoptosis in OGD/R‐treated HAPI cells. Scale bar 50 μm. (E, F) Flow cytometry measured apoptosis rates in OGD/R‐injured HAPI cells after JB treatment (2.5, 5, 10 μM). (G) LDH release assay to measure the effects of JB at concentrations of 2.5, 5, and 10 μM on LDH release in OGD/R‐treated HAPI cells. (H, I) Western blot analysis of the effects of JB (2.5, 5, 10 μM) on the expression of apoptosis‐related proteins (Caspase‐3, Cleaved‐Caspase‐3, Caspase‐9, Cleaved‐Caspase‐9) in OGD/R‐treated HAPI cells. n = 6. *** p < 0.001 vs. Control; # p < 0.05, ## p < 0.05, ### p < 0.001 vs. OGD/R.

Article Snippet: The apoptosis of neuronal cells in rat brain tissue was assessed following the instructions of the TUNEL cell apoptosis detection kit (A113‐01, Vazyme).

Techniques: CCK-8 Assay, TUNEL Assay, Flow Cytometry, Lactate Dehydrogenase Assay, Western Blot, Expressing, Control

Journal: CNS Neuroscience & Therapeutics

Article Title: Jolkinolide B Mitigates Cerebral Ischemia–Reperfusion Injury by Promoting Microglial M1 / M2 Polarization Through the JAK2 / STAT3 Signaling Pathway

doi: 10.1111/cns.70653

Figure Lengend Snippet: JB protects HAPI cells from OGD/R‐triggered inflammatory damage via suppression of JAK2/STAT3 pathway activation. (A, B) Western blot was used to verify the effects of JAK2 activator and inhibitor. (C, D) Immunofluorescence analysis was conducted to examine the effects of JB, WP1066, and BE on the nuclear translocation of p‐STAT3 in OGD/R‐treated HAPI cells. Scale bar 50 μm. (E, F) Western blot assessed JB, WP1066, and BE effects on JAK2/STAT3 pathway proteins (JAK2, p‐JAK2, STAT3, p‐STAT3) in OGD/R‐treated HAPI cells. (G) ELISA was used to assess the effects of JB, WP1066, and BE on the levels of pro‐inflammatory cytokines (IL‐1β, TNF‐α, and IFN‐γ) in OGD/R‐treated HAPI cells. (H, I) Flow cytometry was conducted to analyze the effects of JB, WP1066, and BE on the apoptosis rate of OGD/R‐treated HAPI cells. (J) LDH release assay assessed JB, WP1066, and BE effects on OGD/R‐treated microglial cells. n = 6. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. Control; ### p < 0.001 vs. OGD/R; & p < 0.05, &&& p < 0.001 vs. OGD/R + JB.

Article Snippet: The apoptosis of neuronal cells in rat brain tissue was assessed following the instructions of the TUNEL cell apoptosis detection kit (A113‐01, Vazyme).

Techniques: Activation Assay, Western Blot, Immunofluorescence, Translocation Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Lactate Dehydrogenase Assay, Control