Journal: Frontiers in Pharmacology

Article Title: Enhanced therapeutic potential of paeoniflorin and vitamin B12 in intracerebropeduncle ethidium bromide-induced multiple sclerosis-like pathology

doi: 10.3389/fphar.2026.1792674

Figure Lengend Snippet: (A–D) PNN neuroprotective role in mitigating EBRO-induced alterations in levels of pro-inflammatory cytokines like TNF-α (A) , IL-1β (B) , IL-10 (C) in brain homogenates and TNF-α, IL-1β, and IL-10 in blood plasma (D) . To identify significant differences between groups, a one-way ANOVA and Tukey’s post hoc test were used for statistical analysis (A–D) . The statistical significance level was set at p < 0.01, and the data were displayed as mean ± standard deviation (SD). There were eight wistar rats (n = 8) in each experimental group. β v/s Sham Control, Vehicle Control, and PNN Perse; δ v/s EBRO; δα1 v/s EBRO + PNN50; δα2 v/s EBRO + PNN100, EBRO + PNN50; and δα3 v/s EBRO + VB12 (30), EBRO + PNN100, EBRO + PNN50.

Article Snippet: Also assessed neuroinflammatory cytokines TNF-alpha [KB1145; Krishgen Biosystem, Mumbai, India] and IL-1 Beta [KLR0119; Krishgen Biosystem, Mumbai, India] , and IL-10 [GENLISA, Krishgen, Maharashtra, India] ( ; ).

Techniques: Clinical Proteomics, Standard Deviation, Control

Journal: Frontiers in Pharmacology

Article Title: Enhanced therapeutic potential of paeoniflorin and vitamin B12 in intracerebropeduncle ethidium bromide-induced multiple sclerosis-like pathology

doi: 10.3389/fphar.2026.1792674

Figure Lengend Snippet: (A–H) Neuroprotection by PNN reduces LFB-stained histopathological changes in coronal section-cerebral cortex in rat model of MS. All images displayed on the Micro-LFB-Panel were taken at a ×40 magnification of cortical coronal sections of adult Wistar rats treated with PNN stained. LFB staining is specifically used to examine myelination; it stains the white matter blue and is employed to detect demyelination and remyelination. The experimental groups included (A) Sham Control, (B) Vehicle Control, (C) PNN Perse, (D) EBRO, (E) EBRO + PNN50, (F) EBRO + PNN100, (G) EBRO + VB12 (30), (H) EBRO + VB12 + PNN100. The (Micro-LFB-Panel-A-B-C) group showed normal myelination with elongation, as shown by a black trigon, and white matter tracts under the grey matter showed healthy myelin sheaths. Neurons were packed densely, and almost no vacuole formation was shown by the orange trigon, and no damage to the white matter. (Micro-LFB-Panel D) The EBRO group exhibited extensive demyelination; loss of blue staining in the LFB indicated severe damage to the myelin sheaths, with very prominent dead macrophages (pink trigon) and myelin breakdown products, vacuole formation indicative of tissue degeneration, and a spongy appearance in the affected areas. (Micro-LFB-Panel-E) The EBRO + PNN50 group shows moderate reductions in demyelination, with partial remyelination evident by restoration of LFB staining. (Micro-LFB-Panel-F) In the EBRO + PNN100 group, myelin recovery was much more substantial, associated with robust LFB staining and well-organized myelinated tracts. The neuroinflammatory markers reveal a significant reduction in vacuoles and macrophages, indicating the potency of high-dose PNN for promoting remyelination and controlling neuroinflammation. (Micro LFB Panel-G) in the EBRO + VB12 (30) group, which protects and repairs the myelin sheath. (Micro-LFB-Panel-H) In the EBRO + VB12 (30) +PNN100 combinations, with extensive remyelination and recovery of white matter integrity. The LFB staining intensity in this group is comparable to that of the control groups, and vacuolar size is markedly reduced. Macrophage infiltration is almost absent, indicating that neuroinflammation is efficiently controlled. This finding shows that the combined administration of VB12 (30) + PNN100 had a therapeutic effect, reducing demyelination and restoring myelin structural integrity in the cortex. PNN dosage, with higher doses being much more capable of repairing EBRO-induced white matter damage and restoring normal cortical functions. (Note: The black cube arrow symbolizes myelination/demyelination; the pink cube arrow shows macrophage infiltration; the orange cube arrow indicates vacuolization). (Magnification = ×40; Scale bar = 50 μm).

Article Snippet: Also assessed neuroinflammatory cytokines TNF-alpha [KB1145; Krishgen Biosystem, Mumbai, India] and IL-1 Beta [KLR0119; Krishgen Biosystem, Mumbai, India] , and IL-10 [GENLISA, Krishgen, Maharashtra, India] ( ; ).

Techniques: Staining, Control

Journal: Frontiers in Pharmacology

Article Title: Enhanced therapeutic potential of paeoniflorin and vitamin B12 in intracerebropeduncle ethidium bromide-induced multiple sclerosis-like pathology

doi: 10.3389/fphar.2026.1792674

Figure Lengend Snippet: (A–B) Neuroprotection by PNN reduces LFB-stained histopathological changes in coronal section-hippocampus in rat model of MS. All the images presented in the Micro-LFB-Panel were made at a magnification of ×40 from the coronal sections of the hippocampus in adult Wistar rats treated with PNN stained. LFB staining is used exclusively to examine myelination; it stains white matter blue and has noted to show demyelination and remyelination. Experimental groups were as follows: (A) Sham Control, (B) Vehicle Control, (C) PNN Perse, (D) EBRO, (E) EBRO + PNN50, (F) EBRO + PNN100, (G) EBRO + VB12 (30), (H) EBRO + VB12 + PNN 100. (Micro-LFB-Panel-A-B-C) groups exhibited normal myelinated tracks depicted with the black trigon, and white matter tracts under the grey matter showed healthy myelin sheaths. Neurons were densely packed, showing minimal vacuole formation (shown by the orange trigon) as the entire white matter remained undamaged. (Micro-LFB-Panel D) The EBRO group exhibited massive demyelination, with the loss of blue staining in LFB indicating serious damage to the myelin sheaths, plus numerous heavily loaded macrophages as observed by the pink trigon; the products of myelin breakdown are associated with vacuole formation, denoting tissue degeneration while causing a spongy appearance in affected areas. (Micro-LFB-Panel-E) In the EBRO + PNN50 group, there is a moderate decrease in demyelination compared to the EBRO group. The restoration of the LFB staining indicates partial remyelination. (Micro-LFB-Panel-F) In the EBRO + PNN100 group, myelin recovery was much more robust with LFB staining and well-organised myelinated tracts. The neuroinflammatory markers show a significant reduction in vacuoles and macrophages, indicating that high-dose PNN exerts a potent effect on remyelination and neuroinflammatory control. (Micro-LFB-Panel-G) The EBRO + VB12 (30) group represents protection and repair of the myelin sheath. (Micro-LFB-Panel-H) In the EBRO + VB12 (30) + PNN100 combinations, extensive remyelination and recovery of white matter integrity. In this group, the LFB staining intensity is almost the same as that of the control groups, and vacuole size is markedly decreased. Macrophage infiltration is minimal, indicating that neuroinflammation is effectively controlled. This finding shows that the combined administration in the VB12 (30) + PNN100 group had a therapeutic effect, decreasing demyelination and restoring myelin structural integrity in the hippocampus. PNN100 dosage, with higher doses being much more capable of repairing EBRO-induced white matter damage and restoring normal hippocampal functions. (Note: The black cube symbolizes myelination/demyelination; the pink cube shows macrophage infiltration; the orange cube indicates vacuolization). (Magnification = ×40; Scale bar = 50 μm).

Article Snippet: Also assessed neuroinflammatory cytokines TNF-alpha [KB1145; Krishgen Biosystem, Mumbai, India] and IL-1 Beta [KLR0119; Krishgen Biosystem, Mumbai, India] , and IL-10 [GENLISA, Krishgen, Maharashtra, India] ( ; ).

Techniques: Staining, Control

Journal: Frontiers in Pharmacology

Article Title: Enhanced therapeutic potential of paeoniflorin and vitamin B12 in intracerebropeduncle ethidium bromide-induced multiple sclerosis-like pathology

doi: 10.3389/fphar.2026.1792674

Figure Lengend Snippet: (A–B) Neuroprotection by PNN reduces LFB-stained histopathological changes in the coronal section of the striatum in the rat model of MS. All the images presented in the Micro-LFB-Panel were made at a magnification of ×40 from the coronal sections of the striatum in adult Wistar rats treated with PNN staining. LFB staining is used exclusively for the examination of myelination; it stains white matter blue and notes demyelination and remyelination. The experimental groups included (A) Sham control, (B) Vehicle control, (C) PNN Perse, (D) EBRO, (E) EBRO + PNN50, (F) EBRO + PNN100, (G) EBRO + VB12 (30), and (H) EBRO + VB12 (30) + PNN100. (Micro-LFB-Panel-A-B-C) groups exhibited normal myelinated tracks, followed by the black trigon, and the myelinated fibers right under the grey matter showed healthy myelin sheaths. Neurons clustered together finely, showing very few vacuoles formed (shown by the orange trigon), as the remaining white matter. (Micro-LFB-Panel-D) In the neurotoxin EBRO, more demyelination and the complete loss of blue staining in LFB were observed, revealing severe damage to the myelin sheaths and macrophages seen as the pink trigon. Myelin breakdown products are associated with the formation of vacuoles, signifying tissue degeneration and creating a spongy appearance in affected regions. (Micro-LFB-Panel-E) The EBRO + PNN50 group showed significantly less demyelination than the EBRO group, with partial restoration of LFB staining indicating remyelination. (Micro-LFB-Panel-F) In the EBRO + PNN100 group, myelin recovery was much higher, with robust LFB staining and finely organized myelinated tracks. The neuroinflammatory markers showed a significant reduction in vacuoles and macrophages, indicating that high-dose PNN packs support remyelination and control neuroinflammation. (Micro-LFB-Panel-G) The EBRO + VB12 (30) group indicates that protection and repair of the myelin sheath occur. This group remyelinated and recovered white matter structural integrity; LFB staining intensity was approximately equal to that of controls, with a marked reduction in vacuole size. Furthermore, macrophage infiltration is nearly absent, indicating that neuroinflammation is controlled. (Micro-LFB-Panel-H) VB12 (30) + PNN100 group therapeutic effect through moderating demyelination and regaining the structural integrity of myelin in the striatum. Higher doses of PNN100 are much better than the lower ones in repairing EBRO-induced white matter damage and returning striatal functions to normal. (Note: The black cube symbolizes myelination/demyelination; the pink cube shows macrophage infiltration; the orange cube indicates vacuolization). (Magnification = ×40; Scale bar = 50 μm).

Article Snippet: Also assessed neuroinflammatory cytokines TNF-alpha [KB1145; Krishgen Biosystem, Mumbai, India] and IL-1 Beta [KLR0119; Krishgen Biosystem, Mumbai, India] , and IL-10 [GENLISA, Krishgen, Maharashtra, India] ( ; ).

Techniques: Staining, Control

Journal: Frontiers in Pharmacology

Article Title: Enhanced therapeutic potential of paeoniflorin and vitamin B12 in intracerebropeduncle ethidium bromide-induced multiple sclerosis-like pathology

doi: 10.3389/fphar.2026.1792674

Figure Lengend Snippet: (A–H) Neuroprotection by PNN reduces LFB-stained histopathological changes in coronal section-midbrain in rat model of MS. All the images that are represented in the Micro-LFB-Panel were made at a magnification of ×40 from the coronal sections of the adult Wistar rat midbrain with PNN staining. LFB staining is for the examination of only myelination; it stains white matter blue while advising on demyelination and remyelination. The experimental groups were: (A) Sham Control, (B) Vehicle Control, (C) PNN Perse, (D) EBRO, (E) EBRO + PNN50, (F) EBRO + PNN100, (G) EBRO + VB12 (30), (H) EBRO + VB12 + PNN100. (Micro-LFB-Panel-A-B-C) groups showed normal myelinated tracks depicted with the black trigon, and myelin sheaths under diseased grey matter were normal. The neurons were packed so tightly, with less cellular vacuolation (orange trigon), that the entire white matter remained intact. (Micro-LFB-Panel D) In the EBRO group, extensive demyelination was observed, alongside the absence of blue staining in LFB; this was highly indicative of myelin sheath damage, shown by pink trigons showing numerous heavily-loaded macrophages; breakdown of myelin was found clustered and causing vacuolation, indicating degeneration of affected tissue and a spongy appearance. (Micro-LFB-Panel-E) The EBRO + PNN50 group showed a moderate decrease in demyelination compared with the EBRO group. (Micro-LFB-Panel-F) In the EBRO + PNN100 group, myelin recovery was seen as extremely high LFB staining intensity and well-organised myelinated tracts. As indicated by neuroinflammatory markers, fewer vacuoles and macrophages were observed, suggesting that high-dose PNN has a potent effect on remyelination and on inhibiting neuroinflammation. (Micro-LFB-Panel-G) The EBRO + VB12 (30) group signifies repair and protection of the myelin sheath. (Micro-LFB-Panel-H) The EBRO + VB12 (30) + PNN100 combination denotes widespread remyelination and repair of the integrity of white matter. In this group, LFB staining intensity approaches that of the control groups, with markedly reduced vacuolar sizes. Macrophage infiltration is a clear indication that neuroinflammation is effectively contained. VB12 (30) + PNN100 has a great therapeutic potential for reducing demyelination and restoring the integrative structure of the midbrain. (Note: The black cube symbolizes myelination/demyelination; the pink cube shows macrophage infiltration; the orange cube indicates vacuolization). (Magnification = ×40; Scale bar = 50 μm).

Article Snippet: Also assessed neuroinflammatory cytokines TNF-alpha [KB1145; Krishgen Biosystem, Mumbai, India] and IL-1 Beta [KLR0119; Krishgen Biosystem, Mumbai, India] , and IL-10 [GENLISA, Krishgen, Maharashtra, India] ( ; ).

Techniques: Staining, Control

Journal: Frontiers in Pharmacology

Article Title: Enhanced therapeutic potential of paeoniflorin and vitamin B12 in intracerebropeduncle ethidium bromide-induced multiple sclerosis-like pathology

doi: 10.3389/fphar.2026.1792674

Figure Lengend Snippet: (A–H) Neuroprotection by paeoniflorin reduces LFB-stained histopathological changes in coronal section-cerebellum in rat model of multiple sclerosis. All images presented in the Micro-LFB-Panel were captured at a magnification of ×40, using coronal sections from the cerebellum of adult Wistar rats. This LFB staining technique is primarily used to examine myelination, staining white matter blue and highlighting demyelination and remyelination. The experimental groups were categorized as follows: (A) Sham Control, (B) Vehicle Control, (C) PNN Perse, (D) EBRO, (E) EBRO + PNN50, (F) EBRO + PNN100, (G) EBRO + VB12 (30), and (H) EBRO + VB12 (30) + PNN100. (Micro-LFB-Panel-A-B-C) group exhibited normal myelinated tracks as indicated by black trigons. Additionally, the underlying white and grey matter showed intact myelin sheaths. Neurons appeared densely packed with minimal vacuole formation, as marked by orange trigons; overall, the white matter remained undamaged. (Micro-LFB-Panel-D) group, significant demyelination was observed in the EBRO group. The loss of blue coloration in LFB highlighted severe damage to myelin sheaths and numerous macrophages in affected regions—this association was shown by pink trigons. (Micro-LFB-Panel-E) The EBRO + PNN50 group showed a smaller decrease in demyelination than the EBRO group. (Micro-LFB-Panel-F) The EBRO + PNN100 group closely reveals a more robust recovery characterized by intense LFB staining and well-organized myelinated tracts. Neuroinflammatory markers showed a notable decrease in both vacuoles and macrophages, and a high dose of PNN promoted remyelination while effectively managing neuroinflammation. (Micro-LFB-Panel-G) The EBRO + VB12 (30) group exhibits protection and repair of the myelin sheath. (Micro-LFB-Panel-H) The EBRO + VB12 (30) + PNN100 combination shows extensive remyelination and recovery of white matter integrity. In this group, the LFB staining density is comparable to that of the control groups with reduced vacuolar size. Macrophage infiltration appears to be absent. This result shows that the combination of VB12 (30) + PNN100 has a strong therapeutic effect in decreasing demyelination and restoring myelin structural integrity in the cerebellum. PNN dosage should be more effective in restoring EBRO-induced white matter damage and restoring cerebellar function to normal levels. (Note: The black cube arrow symbolizes myelination/demyelination; the pink cube arrow shows macrophage infiltration; the orange cube arrow indicates vacuolization) (Magnification = ×40; Scale bar = 50 μm).

Article Snippet: Also assessed neuroinflammatory cytokines TNF-alpha [KB1145; Krishgen Biosystem, Mumbai, India] and IL-1 Beta [KLR0119; Krishgen Biosystem, Mumbai, India] , and IL-10 [GENLISA, Krishgen, Maharashtra, India] ( ; ).

Techniques: Staining, Control