This study evaluated the immunogenicity of the Bacillus Calmette-Guérin (BCG) vaccine in a guinea pig model to refine preclinical assessment methods. 24 guinea pigs were divided into four groups for immunohistochemical, histopathological, and molecular analyses, including qRT-PCR and ELISA. The ELISA results revealed significant elevations in interleukin 2 (IL-2), interferon-gamma (IFN- ), and tuberculosis-specific antibodies in vaccinated guinea pigs, particularly γ notable after 6 weeks. Although lung cytokine levels remained unchanged, spleen gene expression showed significant differences in interleukin-17, interleukin-12, interleukin-1β, and C-X-C motif chemokine ligand 10 after 6 weeks. Immunohistochemistry revealed peak IL-2 expression at 8 weeks and significant IFN-γ and TNF-α expression at 6 weeks. This study confirmed the effectiveness of BCG vaccine in guinea pigs, providing crucial insights for future tuberculosis vaccine development and standardizing immune response indicators.

Cerebral ischemia is a serious neurological disorder that can lead to high morbidity and mortality. Baicalin is a naturally bioactive flavonoid derived from Scutellaria baicalensis Georgi, which has neuroprotective activity. Baicalin exerts a neuroprotective effect against hypoxic ischemic injury. In this study, we investigated whether baicalin regulates specific proteins in the cerebral cortex of ischemic stroke animals. Middle cerebral artery occlusion (MCAO) surgery was performed to induce ischemic brain injury, and baicalin (30 mg/kg) or vehicle was injected into the abdominal cavity before MCAO surgery. Neurological behavior tests were performed 24 h after MCAO surgery and proteomics approach was performed using proteins extracted from cortical tissue. Two-dimensional electrophoresis analysis and MALDI-TOF were performed to identify the regulated protein by baicalin. MCAO damage caused severe behavioral disorders, but baicalin treatment improved these behavioral deficits. Baicalin also induced changes in the expression of various proteins in the cerebral cortex of MCAO animals. Proteins changed by baicalin administration are as follow: adenosylhomocysteinase, isocitrate dehydrogenase [NAD] subunit alpha, apolipoprotein A-I, Rab GDP dissociation inhibitor beta, eukaryotic initiation factor 4A, and mu-crystallin. These proteins were involved in metabolism and protein synthesis. The results of this study demonstrated the neuroprotective effects of baicalin by improving behavioral disorders caused by MCAO damage. The results also showed that baicalin regulates the expression of a variety of proteins involved in neuroprotective functions. Therefore, our findings provide evidence that baicalin plays a neuroprotective role in stroke animal models by regulating specific proteins.

Ischemic stroke is caused by a blockage of the cerebral artery, which leads to a severe neurological disorder. Chlorogenic acid is a phenolic acid found mainly in plants such as coffee beans, eggplants, and carrots. It exerts a neuroprotective effect against cerebral ischemic damage. Bcl-2 family protein is a representative apoptosis regulatory protein. Bcl-2 and Bcl-xL act as apoptosis inhibitors, while Bax and Bad act as apoptosis inducers.The interaction of Bcl-2 family protein plays an important role in determining cell fate. The aim of this study was to investigate whether chlorogenic acid modulates the interaction of Bcl-2 family proteins during ischemic injury. Middle cerebral artery occlusion (MCAO) surgery was performed to induce cerebral ischemia. Chlorogenic acid (30 mg/kg) or phosphate buffered saline was intraperitoneally injected to adult male rats 2 h after MCAO surgery. Neurobehavioral tests were performed to confirm the neuroprotective effect of chlorogenic acid 24 h after MCAO injury, and immunoprecipitation analysis was performed to investigate the interaction of Bcl-2 family protein. MCAO damage showed signs of severe neurological disorders, while chlorogenic acid improved these disorders. Results of immunoprecipitation analysis were as follows. Interaction between Bax and Bcl-2 or Bcl-xL was decreased in MCAO injury, chlorogenic acid prevents these decreases. In contrast to Bax, Interaction between Bad and Bcl-2 or Bcl-xL was increased in MCAO injury, chlorogenic acid prevents these increases. Furthermore, chlorogenic acid attenuated MCAO-induced increase of capase-9. In conclusion, our findings demonstrate that chlorogenic acid exerts a neuroprotective effect against cerebral ischemic injury by modulating interaction of Bcl-2 family proteins.

Ischemic stroke leads to severe brain damage and high mortality. Chlorogenic acid is a phenolic compound known to have neuroprotective properties. Bcl-2 family protein plays an important role in the regulation of apoptosis. We investigated whether chlorogenic acid exerts neuroprotective effects against ischemic injury by modulating Bcl-2 and Bax proteins. Middle cerebral artery occlusion (MCAO) was performed to induce cerebral ischemia and rats were injected intraperitoneally with phosphate buffered saline or chlorogenic acid (30 mg/kg) for 2 h after MCAO. Cortical tissues were collected 24 h after MCAO injury and reverse transcription-quantitative real time polymerase chain reaction and Western blot analyses were performed to investigate the expression of Bcl-2 and Bax. The regulation of Bcl-2 and Bax proteins by chlorogenic acid during glutamateinduced cell damage were examined. Cells were collected at 24 h after administration of glutamate (5 mM) and chlorogenic acid (10, 30, 50 μM). These results showed a decrease in Bcl-2 expression and an increase in Bax expression in MCAO animals, but chlorogenic acid treatment alleviated these changes by MCAO damage. Glutamate significantly reduced cell viability, and chlorogenic acid treatment alleviated this reduction in a dose-dependent manner. Glutamate induced a decrease in Bcl-2 expression and an increase in Bax expression, but chlorogenic acid treatment alleviated these changes. We found that chlorogenic acid alleviates changes in the expression of Bcl-2 and Bax proteins induced by brain injury. Therefore, our findings provide an evidence that chlorogenic acid has neuroprotective effects against MCAO damage by modulating Bcl-2 and Bax proteins.

Ischemic stroke causes severe neuronal damage. Chlorogenic acid is a phenolic substance present in fruits and coffee. It also exerts neuroprotective effects against various brain injuries. The 14-3-3 family protein perform a variety of functions including metabolism, signal transduction, cell differentiation, and apoptosis. The purpose of this study is to investigate whether chlorogenic acid regulates the expression of 14-3-3 protein in stroke animal models. Ischemic stroke was induced by middle cerebral artery occlusion (MCAO) surgery. Phosphate buffered saline (PBS) or chlorogenic acid (30 mg/kg) were intraperitoneally injected to adult male rats 2 h before MCAO surgery. Adhesive-removal test was performed 24 h after MCAO surgery and cerebral cortical tissues were collected for further study. MCAO damage caused severe neurological impairment and chlorogenic acid treatment ameliorated this disorder. Our proteomic approach showed a decrease in 14-3-3 expression in MCAO animals with PBS. The decrease in 14-3-3 expression alleviated in MCAO animal with chlorogenic acid. We confirmed changes in various 14-3-3 protein isoforms, including beta/alpha, zeta/delta, gamma, epsilon, eta, and tau through reverse transcription-PCR. These results explained that chlorogenic acid regulates the expression of 14-3-3 protein in MCAO-induced cerebral ischemia. 14-3-3 is considered to be an important protein for cell survival through binding to pro-apoptotic proteins. The maintenance of 14-3-3 levels is an important event in neuroprotection against ischemic injury. Therefore, we can demonstrate that the 14-3-3 protein contributes to the neuroprotective effect of chlorogenic acid in stroke animal models.

Ischemic stroke causes brain damage and neuronal cell death by depriving oxygen and nutrients and releasing excessive levels of glutamate and intracellular calcium. Epigallocatechin gallate (EGCG) is a polyphenolic compound present in green tea. It has antioxidant, anti-inflammatory, and neuroprotective effects. Hippocalcin is a calcium binding protein that regulates calcium concentration, neuronal differentiation, neuronal excitability, and neuronal cell death. In this study, we investigated whether EGCG regulates the expression of hippocalcin in neurons and astrocytes after focal cerebral ischemia. Cerebral ischemia was induced by meddle cerebral artery occlusion (MCAO). EGCG (50 mg/kg) or PBS was injected into the abdominal cavity just before MCAO surgery. Neurobehavioral tests were performed to evaluate the effect of EGCG on neurological behavioral deficits 24 h after MCAO surgery. Immunofluorescence staining was performed to evaluate the positive response to hippocalcin in the cerebral cortex after MCAO surgery. We also detected the positive reactions of neuronal nuclear protein (NeuN) and glial fibrillary acidic protein (GFAP) as markers of neuron and astrocyte, respectively. MCAO caused severe neurological impairment and EGCG treatment attenuated these impairments. MCAO damage reduced the number of NeuN-positive cells and increased the number of GFAP-positive cells. This result indicates a decrease in neurons and an increase in astrocytes. However, EGCG alleviated these changes caused by MCAO damage. MCAO reduced the number of hippocalcin-positive cells in neurons and astrocytes, and EGCG treatment attenuated these reductions. Hippocalcin exerts neuroprotective effect through regulating intracellular calcium concentration. In conclusion, EGCG regulates the expression of hippocalcin in neurons and astrocytes and has neuroprotective effects in focal cerebral ischemia.

Baicalin is a flavonoid compound with many advantages, including anti-inflammatory agents and antioxidants. Lipopolysaccharide (LPS) is an endotoxin that induces neuronal damage through inflammatory response and oxidative stress reaction. This study was investigated the protective effects of baicalin on the oxidative stress and histopathological changes caused by LPS in hippocampus. Adult mice were divided into four groups; vehicle-treated, baicalin-treated, LPS-treated, and LPS and baicalin co-treated animals. Baicalin (10 mg/kg/day) and/or LPS (250 μg/kg/day) were intraperitoneally administered for seven consecutive days, and body weight was measured. Reactive oxygen species (ROS) level and lipid peroxidation level in the hippocampus were examined. Histopathological study was performed using hematoxylin and eosin staining manuals. LPS treatment decreased body weight and increased ROS and oxidative stress in the hippocampus. However, co-treatment with baicalin alleviated these changes caused by LPS. Severe histopathological changes were observed in the hippocampus of LPS-treated animals. Baicalin co-treatment attenuated the changes and preserved neuronal cells from LPS damage. These results showed that baicalin suppresses LPS-induced neuronal damage by alleviating oxidative stress in the hippocampus. Thus, this study demonstrated that baicalin exerts protective effects against LPS-induced oxidative stress in hippocampus.

Lipopolysaccharide (LPS)는 염증유발 cytokine 분비를 자극하고 염증을 유발하는 그람음성균의 내독소이다. 본 연구에서는 LPS가 신경아교세포 활성과 해마에 있는 nuclear factor kappa B (NF-κB) 매개 염증유발 요소를 조절하는지를 조사하였다. 성체 수컷 쥐를 대조군과 LPS를 투여한 실험군으로 무작위로 나누어 vehicle 또는 LPS (250 μg/kg)를 5일 동안 복강투여하고 무게를 측정했다. 해마의 활성산소와 과산화지방질 수준을 분석하고, 형태학적 연구를 위해 Hematoxylin and eosin 염색을 시행하였다. 또한, 해마에서 ionized calcium-binding adapter molecule 1 (Iba-1), glial fibrillary acidic protein (GFAP), NF-κB, interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α)의 발현을 확인하기 위해 Western blot 분석과 immunofluorescence 염색을 시행하였다. 그 결과, LPS를 투여한 쥐들의 체중이 감소하였다. LPS 투여는 활성산소와 과산화지방질 수준의 증가를 유발하였고 LPS를 투여한 쥐의 해마에서 심각한 조직병리학적 변화를 확인했다. 또한 LPS 투여는 신경교세포와 별아교세포의 표시물인 Iba-1과 GFAP의 발현을 증가시켰고, NF-κB의 발현과 IL-1β와 TNF-α와 같은 염증성인자의 발현을 증가시켰다. 이러한 결과들을 통해 LPS 투여는 해마손상과 염증반응을 유도한다는 것을 알 수 있고 LPS 투여가 해마조직에서 신경아교세포와 NF-κB에 매개된 염증인자들을 활성화시킨다는 것을 확인하였 다. 따라서, 본 연구는 LPS 투여는 해마조직에서 산화적 스트레스 증가와 염증인자 활성을 증가시켜 신경손상을 유도함을 보여준다.

Curcumin plays a protective role in brain injury through its anti-oxidant and anti-inflammatory activities. Moreover, peroxiredoxin-5 exerts a protective effect against oxidative stress. The aim of this study was to investigate whether curcumin modulated the peroxiredoxin-5 expression in focal cerebral ischemic animal model. Middle cerebral artery occlusion(MCAO) was performed to induce cerebral ischemic injury in rats. Adult male rats were injected intraperitoneally with vehicle or curcumin(50㎎/㎏ B.W.) 1 h after MCAO and cerebral cortex tissues were collected 24 h after MCAO. Photographs of hematoxylin and eosin staining showed that MCAO induced necrotic changes with scalloped shrunken form and apoptotic changes with nuclear chromatin condensations. However, curcumin treatment attenuated MCAO-induced histopathological changes. Moreover, this study clearly showed that peroxiredoxin-5 expression was decreased in MCAO operated animal with vehicle using a proteomics approach. However, this decrease in peroxiredoxin-5 expression was attenuated by curcumin treatment. Reverse-transcription PCR and Western blot analyses confirmed that curcumin treatment alleviated the MCAO injury-induced decrease in peroxiredoxin-5 expression(p<0.05). These results demonstrated that curcumin regulates peroxiredoxin-5 expression in MCAO animal model. In conclusion, our findings suggest that curcumin exerts a neuroprotective effect in cerebral ischemia by attenuating the MCAO-induced decrease in peroxiredoxin-5 expression.

Calbindin-D28k is a calcium-binding protein that mediates intracellular calcium concentrations and exerts a neuroprotective effect against ischemic injury. Ferulic acid provides a neuroprotective effect against focal cerebral ischemia through its anti-oxidative and anti-inflammatory mechanisms. In this study, we investigated whether ferulic acid regulates calbindin-D28k expression during focal cerebral ischemia and glutamate treatment-induced neuronal cell death. Middle cerebral artery occlusion (MCAO) was performed to induce focal cerebral ischemia. Ferulic acid (100 mg/kg, i.v.) or vehicle was immediately administered after MCAO, and brain tissues were isolated 24 h after MCAO. RT-PCR and Western blot analyses showed a decrease in calbindin-D28k in MCAO-operated animals. We found that ferulic acid treatment prevented the MCAO-induced decrease in calbindin-D28k expression. Glutamate exposure elevated the intracellular calcium levels in cultured hippocampal cells, and ferulic acid prevented the glutamate exposure-induced increase in calcium levels. Moreover, ferulic acid also attenuated the glutamate toxicity-induced decrease in calbindin-D28k. Taken together, these in vivo and in vitro results demonstrate that ferulic acid regulates calbindin-D28k expression in neuronal cell injury. Therefore, these findings suggest that ferulic acid exerts a neuroprotective effect by modulating calbindin-D28k expression.

본 연구는 중간대뇌동맥을 폐쇄한 대뇌허혈성 손상모델에서 ferulic acid에 의해 조절되는 HO-1과 HO-2의 발현에 관하여 조사하였다. 흰쥐(Sprague-Dawley, 수컷)에 ferulic acid (100 mg/kg) 또는 vehicle을 중간대뇌동맥폐쇄술(MCAO) 후 정맥으로 주사하였고 중간대뇌동맥폐쇄술(MCAO)을 실시한 24시간 후 대뇌피질의 조직을 적출하였다. Hematoxylin과 eosin 염색을 통하여 MCAO로 유도된 뇌 손상시 ferulic acid의 보호효과를 확인하였다. MCAO을 시행한 대뇌피질에서는 응축된 핵과 신경세포의 괴사 소견을 보였으나, ferulic acid 투여군에서는 이들 신경세포의 병변을 현저히 완화시켰다. HO-1과 HO-2의 RNA와 단백질 발현의 변화를 reverse-transcription PCR과 Western blot으로 분석하였다. HO-1 발현은 MCAO 후 vehicle 투여군에서 현저히 감소하였으나, MCAO 후 ferulic acid를 투여한 실험군에서는 이들 감소의 완화를 보였으며, MCAO를 시행하지 않은 실험군의 수준으로 유지되었다. 그러나, HO-2의 발현은 MCAO 후 vehicle 투여군과 ferulic acid 투여군에서 유의적인 차이는 관찰되지 않았고 MCAO를 시행하지 않은 실험군의 수준으로 유지되었다. 따라서, 본 연구의 결과는 허혈성 뇌 손상시 ferulic acid는 HO-1 발현을 조절하였으나, HO-2의 발현에는 영향을 미치지 못함을 확인하였다. 결론적으로, 허혈성 뇌손상시 ferulic acid는 HO-1의 발현을 조절하여 신경세포를 보호하는 역할을 수행한다는 사실을 확인하였다.