OBJECTIVE: Treating peripheral nerve injury (PNI) presents a clinical challenge due to limited axon regeneration. Strychni Semen, a traditional Chinese medicine, is clinically used for numbness and hemiplegia. However, its role in promoting functional recovery after PNI and the related mechanisms have not yet been systematically studied. METHODS: A mouse model of sciatic nerve crush (SNC) injury was established and the mice received drug treatment via intragastric gavage, followed by behavioral assessments (adhesive removal test, hot-plate test and Von Frey test). Transcriptomic analyses were performed to examine gene expression in the dorsal root ganglia (DRGs) from the third to the sixth lumbar vertebrae, so as to identify the significantly differentially expressed genes. Immunofluorescence staining was used to assess the expression levels of superior cervical ganglia neural-specific 10 protein (SCG10). The ultra-trace protein detection technique was used to evaluate changes in gene expression levels. RESULTS: Strychni Semen and its active compounds (brucine and strychnine) improved functional recovery in mice following SNC injury. Transcriptomic data indicated that Strychni Semen and its active compounds initiated transcriptional reprogramming that impacted cellular morphology and extracellular matrix remodeling in DRGs after SNC, suggesting potential roles in promoting axon regeneration. Imaging data further confirmed that Strychni Semen and its active compounds facilitated axon regrowth in SNC-injured mice. By integrating protein-protein interaction predictions, ultra-trace protein detection, and molecular docking analysis, we identified myeloperoxidase as a potentially critical factor in the axon regenerative effects conferred by Strychni Semen and its active compounds. CONCLUSION Strychni Semen and its active compounds enhance sensory function by promoting axonal regeneration after PNI. These findings establish a foundation for the future applications of Strychni Semen and highlight novel therapeutic strategies and drug targets for axon regeneration. Please cite this article as: Zhang Y, Zhao XY, Liu MT, Zhou ZC, Cheng HB, Jiang XH, Zheng YR, Chen Z. Strychni Semen and its active compounds promote axon regeneration following peripheral nerve injury by suppressing myeloperoxidase in the dorsal root ganglia. J Integr Med. 2025; 23(2): 169-181.
Animals ; Nerve Regeneration/drug effects* ; Mice ; Peripheral Nerve Injuries/physiopathology* ; Male ; Ganglia, Spinal/enzymology* ; Axons/physiology* ; Peroxidase/antagonists & inhibitors* ; Mice, Inbred C57BL ; Drugs, Chinese Herbal/pharmacology* ; Disease Models, Animal ; Strychnine/pharmacology*

BACKGROUNDIn addition to neurons, all components of the neurovascular unit (NVU), such as glial, endothelial, and basal membranes, are destroyed during traumatic brain injury (TBI). Previous studies have shown that excessive stimulation of calpain is crucial for cerebral injury after traumatic insult. The objective of this study was to investigate whether calpain activation participated in NVU disruption and edema formation in a mouse model of controlled cortical impact (CCI).

METHODSOne hundred and eight mice were divided into three groups: the sham group, the control group, and the MDL28170 group. MDL28170 (20 mg/kg), an efficient calpain inhibitor, was administered intraperitoneally at 5 min, 3 h, and 6 h after experimental CCI. We then measured neurobehavioral deficits, calpain activity, inflammatory mediator levels, blood-brain barrier (BBB) disruption, and NVU deficits using electron microscopy and histopathological analysis at 6 h and 24 h after CCI.

RESULTSThe MDL28170 treatment significantly reduced the extent of both cerebral contusion (MDL28170 vs. vehicle group, 16.90 ± 1.01 mm΃ and 17.20 ± 1.17 mm΃ vs. 9.30 ± 1.05 mm΃ and 9.90 ± 1.17 mm΃, both P < 0.001) and edema (MDL28170 vs. vehicle group, 80.76 ± 1.25% and 82.00 ± 1.84% vs. 82.55 ± 1.32% and 83.64 ± 1.25%, both P < 0.05), improved neurological scores (MDL28170 vs. vehicle group, 7.50 ± 0.45 and 6.33 ± 0.38 vs. 12.33 ± 0.48 and 11.67 ± 0.48, both P < 0.001), and attenuated NVU damage resulting (including tight junction (TJ), basement membrane, BBB, and neuron) from CCI at 6 h and 24 h. Moreover, MDL28170 markedly downregulated nuclear factor-κB-related inflammation (tumor necrosis factor-α [TNF-α]: MDL28170 vs. vehicle group, 1.15 ± 0.07 and 1.62 ± 0.08 vs. 1.59 ± 0.10 and 2.18 ± 0.10, both P < 0.001; inducible nitric oxide synthase: MDL28170 vs. vehicle group, 4.51 ± 0.23 vs. 6.23 ± 0.12, P < 0.001 at 24 h; intracellular adhesion molecule-1: MDL28170 vs. vehicle group, 1.45 ± 0.13 vs. 1.70 ± 0.12, P < 0.01 at 24 h) and lessened both myeloperoxidase activity (MDL28170 vs. vehicle group, 0.016 ± 0.001 and 0.016 ± 0.001 vs. 0.024 ± 0.001 and 0.023 ± 0.001, P < 0.001 and 0.01, respectively) and matrix metalloproteinase-9 (MMP-9) levels (MDL28170 vs. vehicle group, 0.87 ± 0.13 and 1.10 ± 0.10 vs. 1.17 ± 0.13 and 1.25 ± 0.12, P < 0.001 and 0.05, respectively) at 6 h and 24 h after CCI.

CONCLUSIONSThese findings demonstrate that MDL28170 can protect the structure of the NVU by inhibiting the inflammatory cascade, reducing the expression of MMP-9, and supporting the integrity of TJ during acute TBI.

Animals ; Brain Injuries, Traumatic ; drug therapy ; metabolism ; Calpain ; antagonists & inhibitors ; metabolism ; Dipeptides ; therapeutic use ; Disease Models, Animal ; Glycoproteins ; therapeutic use ; Inflammation ; drug therapy ; metabolism ; Male ; Matrix Metalloproteinase 9 ; metabolism ; Mice ; Mice, Inbred BALB C ; NF-kappa B ; metabolism ; Peroxidase ; antagonists & inhibitors ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

To explore the effective ingredients and mechanism of Ligusticum wallichii in treating brain ischemia. Four brain ischemia-related target proteins were selected in the joint screening for the 45 component in L. wallichii reported in literatures based on molecular docking by reference to the corresponding drugs in the market. According to the docking results, multiple components in L. wallichii, such as phthalides, were superior to the corresponding drugs in the market, suggesting that they may be the major effective components in L. wallichii for treating brain ischemia. The method can be used to study the material base and molecular mechanism of traditional Chinese medicines.
Brain Ischemia ; drug therapy ; Ligusticum ; chemistry ; Medicine, Chinese Traditional ; Molecular Docking Simulation ; Nitric Oxide Synthase Type II ; antagonists & inhibitors ; Peroxidase ; antagonists & inhibitors ; Phytotherapy ; Tumor Necrosis Factor-alpha ; antagonists & inhibitors

OBJECTIVEThe roles of cerebrovascular oxidative stress in vascular functional remodeling have been described in hindlimb-unweighting (HU) rats. However, the underlying mechanism remains to be established.

METHODSWe investigated the generation of vascular reactive oxygen species (ROS), Nox2/Nox4 protein and mRNA levels, NADPH oxidase activity, and manganese superoxide dismutase (MnSOD) and glutathione peroxidase-1 (GPx-1) mRNA levels in cerebral and mesenteric smooth muscle cells (VSMCs) of HU rats.

RESULTSROS production increased in cerebral but not in mesenteric VSMCs of HU rats compared with those in control rats. Nox2 and Nox4 protein and mRNA levels were increased significantly but MnSOD/GPx-1 mRNA levels decreased in HU rat cerebral arteries but not in mesenteric arteries. NADPH oxidases were activated significantly more in cerebral but not in mesenteric arteries of HU rats. NADPH oxidase inhibition with apocynin attenuated cerebrovascular ROS production and partially restored Nox2/Nox4 protein and mRNA levels, NADPH oxidase activity, and MnSOD/GPx-1 mRNA levels in cerebral VSMCs of HU rats.

CONCLUSIONThese results suggest that vascular NADPH oxidases regulate cerebrovascular redox status and participate in vascular oxidative stress injury during simulated microgravit.

Acetophenones ; Animals ; Cerebral Arteries ; metabolism ; Glutathione Peroxidase ; metabolism ; Hindlimb Suspension ; Male ; Membrane Glycoproteins ; metabolism ; Mesenteric Arteries ; metabolism ; Myocytes, Smooth Muscle ; metabolism ; NADPH Oxidase 2 ; NADPH Oxidase 4 ; NADPH Oxidases ; antagonists & inhibitors ; metabolism ; Rats, Sprague-Dawley ; Reactive Oxygen Species ; Superoxide Dismutase ; metabolism

Bacterial biofilms have emerged as potential critical triggers in the pathogenesis of bisphosphonate (BP)-related osteonecrosis of the jaw (ONJ) or BRONJ. BRONJ lesions have shown to be heavily colonized by oral bacteria, most of these difficult to cultivate and presents many clinical challenges. The purpose of this study was to characterize the bacterial diversity in BRONJ lesions and to determine host immune response. We examined tissue specimens from three cohorts (n=30); patients with periodontal disease without a history of BP therapy (Control, n=10), patients with periodontal disease having history of BP therapy but without ONJ (BP, n=5) and patients with BRONJ (BRONJ, n=15). Denaturing gradient gel electrophoresis of polymerase chain reaction (PCR)-amplified 16S rRNA gene fragments revealed less bacterial diversity in BRONJ than BP and Control cohorts. Sequence analysis detected six phyla with predominant affiliation to Firmicutes in BRONJ (71.6%), BP (70.3%) and Control (59.1%). Significant differences (P<0.05) in genera were observed, between Control/BP, Control/BRONJ and BP/BRONJ cohorts. Enzyme-linked immunosorbent assay (ELISA) results indicated that the levels of myeloperoxidase were significantly lower, whereas interleukin-6 and tumor necrosis factor-alpha levels were moderately elevated in BRONJ patients as compared to Controls. PCR array showed significant changes in BRONJ patients with downregulation of host genes, such as nucleotide-binding oligomerization domain containing protein 2, and cathepsin G, the key modulators for antibacterial response and upregulation of secretory leukocyte protease inhibitor, proteinase 3 and conserved helix-loop-helix ubiquitous kinase. The results suggest that colonization of unique bacterial communities coupled with deficient innate immune response is likely to impact the pathogenesis of ONJ.
Actinobacteria ; classification ; Bacteria ; classification ; Bacteroidetes ; classification ; Biofilms ; Bisphosphonate-Associated Osteonecrosis of the Jaw ; immunology ; microbiology ; Bone Density Conservation Agents ; therapeutic use ; Cathepsin G ; analysis ; Cohort Studies ; Down-Regulation ; Female ; Fusobacteria ; classification ; Gram-Negative Bacteria ; classification ; Host-Pathogen Interactions ; immunology ; Humans ; I-kappa B Kinase ; analysis ; Immunity, Innate ; immunology ; Interleukin-6 ; analysis ; Male ; Middle Aged ; Mouth ; immunology ; microbiology ; Myeloblastin ; analysis ; antagonists & inhibitors ; Nod2 Signaling Adaptor Protein ; analysis ; Periodontal Diseases ; microbiology ; Peroxidase ; analysis ; Proteobacteria ; classification ; Tumor Necrosis Factor-alpha ; analysis

OBJECTIVE: To investigate the protective effect of a non-specific NMDA receptor antagonist memantine on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice. METHODS: Healthy male mice were divided into 4 groups: a normal group, a memantine group, an ALI group and a memantine+ALI group. The ALI group was induced by intraperitoneal injection of LPS (10 mg/kg). Memantine (10 mg/kg) was injected intraperitoneally before the injection of LPS to determine the effect of blockade of NMDA receptor in the memantine+ALI group. The lung wet/dry ratio was detected. HE staining was preformed to show the morphological changes in the lung tissue. Myeloperoxidase enzyme (MPO) activity and malondialdehyde (MDA) content in the lung tissue were detected. ELISA was used to detect the tumor necrosis factor-α (TNF-α) content and lactate dehydrogenase (LDH) activity in the bronchoalveolar lavage fluid (BALF). RESULTS: Memantine pretreatment improved the LPS-induced ALI lung tissue morphological changes, reduced their lung wet/dry ratio, the levels of TNF-α and LDH activity in BALF, and also reduced the MPO and MDA content in the lung tissue. CONCLUSION Blockade of NMDA receptors can ameliorate LPS-induced mice ALI.
Acute Lung Injury ; drug therapy ; Animals ; Bronchoalveolar Lavage Fluid ; L-Lactate Dehydrogenase ; metabolism ; Lipopolysaccharides ; Lung ; metabolism ; Male ; Malondialdehyde ; metabolism ; Memantine ; pharmacology ; Mice ; Peroxidase ; metabolism ; Receptors, N-Methyl-D-Aspartate ; antagonists & inhibitors ; Tumor Necrosis Factor-alpha ; metabolism

To investigate the influence of hydrogen sulfide (H₂S) on p38 MAPK signaling pathway during acute lung injury (ALI) caused by lipopolysaccharide (LPS), the rats were randomly divided into six groups: control group, LPS group, LPS + NaHS group, LPS + PPG (cystathionine-γ-lyase inhibitor) group, NaHS group and PPG group. The rats were sacrificed 6 h after injection and lung tissues were obtained. The structure of lung tissues and the number of polymorphonuclear leucocyte (PMN) was observed under optical microscope; the lung myeloperoxidase (MPO) activity, superoxide dismutase (SOD) activity and malondialdehyde (MDA) content were tested; intercellular adhesion molecule-1 (ICAM-1) protein expression changes were detected by immunohistochemical staining; phosphorylated p38 MAPK (p-p38 MAPK) protein expression was detected by Western blotting. The results showed that the lung injury in LPS group was observed, at the same time the MPO activity, the content of MDA, ICAM-1 and p-p38 MAPK protein expressions, the number of PMN were all higher than those in control group (all P < 0.05). Pre-injection of NaHS alleviated the changes induced by LPS, while pre-injection of PPG aggravated those alterations (all P < 0.05). ICAM-1 and p-p38 MAPK protein expressions in lung tissue were positively correlated (r = 0.923, P < 0.01). The results suggest that H2S may reduce LPS-induced ALI through inhibiting the conjugation of p38 MAPK and reducing the expression of ICAM-1.
Acute Lung Injury ; chemically induced ; drug therapy ; Animals ; Hydrogen Sulfide ; pharmacology ; Intercellular Adhesion Molecule-1 ; metabolism ; Lipopolysaccharides ; Lung ; metabolism ; pathology ; MAP Kinase Signaling System ; drug effects ; Malondialdehyde ; pharmacology ; Neutrophils ; Peroxidase ; metabolism ; Phosphorylation ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; pharmacology ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; metabolism

This study is to investigate the effect of fenofibrate on angiotensin II (Ang II)-induced toll-like receptor 4 (TLR4) expression, myeloperoxidase (MPO) activity and expression in murine macrophage line RAW264.7 cells and explore its anti-inflammatory mechanism. TLR4 and MPO mRNA levels were analyzed by RT-PCR, and TLR4 and MPO protein expressions were measured by Western blotting. MPO activity in the cell supernatant was assayed with colorimetry. The results showed that fenofibrate reduced Ang II-induced mRNA and protein expression of TLR4 and inhibited activity, mRNA and protein expression of MPO in RAW264.7 cells in concentration-dependent manner. In addition, TLR4 blocker partially antagonized the effect of Ang II on MPO activity in RAW264.7 cells, and fenofibrate potentiated the inhibitory effect. Meanwhile, fenofibrate significantly suppressed LPS (TLR4 special ligand)-induced MPO activity in RAW264.7 cells. In conclusion, fenofibrate downregulated Ang II-induced TLR4 expression and blocked MPO secretion in RAW264.7 cells via interfering with the TLR4-dependent signaling pathway to alleviate inflammation, which might be one of its novel anti-inflammatory mechanisms.
Angiotensin II ; pharmacology ; Animals ; Anti-Inflammatory Agents ; administration & dosage ; pharmacology ; Cell Line ; Dose-Response Relationship, Drug ; Fenofibrate ; administration & dosage ; pharmacology ; Hypolipidemic Agents ; administration & dosage ; pharmacology ; Lipopolysaccharides ; pharmacology ; Macrophages ; cytology ; metabolism ; Mice ; Peroxidase ; metabolism ; RNA, Messenger ; metabolism ; Signal Transduction ; Toll-Like Receptor 4 ; antagonists & inhibitors ; metabolism

OBJECTIVETo further investigate the neuroprotective effects of five isoflavonoids from Astragalus mongholicus on xanthine (XA)/ xanthine oxidase (XO)-induced injury to PC12 cells.

METHODSPC12 cells were damaged by XA/XO. The activities of antioxidant enzymes, MTT, LDH, and GSH assays were used to evaluate the protection of these five isoflavonoids. Contents of Bcl-2 family proteins were determined with flow cytometry.

RESULTSAmong the five isoflavonoids including formononetin, ononin, 9, 10-dimethoxypterocarpan-3-O-beta-D-glucoside, calycosin and calycosin-7-O-glucoside, calycosin and calycosin-7-O-glucoside were found to inhibit XA/ XO-induced injury to PC12 cells. Their EC50 values of formononetin and calycosin were 0.05 microg/mL. Moreover, treatment with these three isoflavonoids prevented a decrease in the activities of antioxidant enzymes, superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), while formononetin and calycosin could prevent a significant deletion of GSH. In addition, only calycosin and calycosin-7-O-glucoside were shown to inhibit XO activity in cell-free system, with an approximate IC50 value of 10 microg/mL and 50 microg/mL. Formononetin and calycosin had no significant influence on Bcl-2 or Bax protein contents.

CONCLUSIONNeuroprotection of formononetin, calycosin and calycosin-7-O-glucoside may be mediated by increasing endogenous antioxidants, rather by inhibiting XO activities or by scavenging free radicals.

Animals ; Astragalus Plant ; chemistry ; Glucosides ; chemistry ; isolation & purification ; pharmacology ; Glutathione ; metabolism ; Glutathione Peroxidase ; metabolism ; Isoflavones ; chemistry ; isolation & purification ; pharmacology ; PC12 Cells ; Protective Agents ; isolation & purification ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Superoxide Dismutase ; metabolism ; Xanthine Oxidase ; antagonists & inhibitors ; pharmacology

Intestinal ischemia-reperfusion (I/R) is an important event in the pathogenesis of multiple organ dysfunction syndrome (MODS). The aim of this study is to determine the effects of ginsenoside Rb1 on liver injury induced by intestinal I/R in rats. Adult male Wistar rats were randomly divided into four groups: (1) a control, sham-operated group (sham group); (2) an intestinal I/R group subjected to 1 h intestinal ischemia and 2 h reperfusion (I/R group); (3) a group treated with 20 mg/kg ginsenoside Rb1 before reperfusion (Rb1-20 group); and (4) a group treated with 40 mg/kg ginsenoside Rb1 before reperfusion (Rb1-40 group). Liver and intestinal histology was observed. Aspartate aminotransferase (AST), alanine aminotransferase (ALT) level in serum and malondialdehyde (MDA) level in intestinal tissues were measured. Myeloperoxidase (MPO), TNF-alpha, MDA level and immunohistochemical expression of NF-kappa B and intracellular adhesion molecale-1 (ICAM-1) in liver tissues was assayed. In addition, a western blot analysis of liver NF-kappa B expression was performed. Results indicated intestinal I/R induced intestinal and liver injury, which was characterized by increase of AST and ALT in serum, MDA level in intestine, MPO, TNF-alpha and MDA level and ICAM-1 and NF-kappa B expression in the liver tissues. Ginsenoside Rb1 (20, 40 mg/kg) ameliorated liver injury, decreased MPO, TNF-alpha and MDA level, NF-kappa B and ICAM-1 expression in liver tissues. In conclusion, ginsenoside Rb1 ablated liver injury induced by intestinal I/R by inhibiting NF-kappaB activation.
Alanine Transaminase/blood ; Animals ; Aspartate Aminotransferases/blood ; Biological Markers/metabolism ; Ginsenosides/*pharmacology ; Intercellular Adhesion Molecule-1/metabolism ; Intestines/*blood supply/metabolism/pathology ; Ischemia/metabolism/pathology ; Liver/enzymology/pathology ; Liver Diseases/etiology/*pathology ; Male ; Malondialdehyde/metabolism ; NF-kappa B/*antagonists & inhibitors/metabolism ; Peroxidase/metabolism ; Rats ; Rats, Wistar ; Reperfusion Injury/complications/*pathology ; Tumor Necrosis Factor-alpha/metabolism