Choi, Chul-Hoon
(Department of Acupuncture & Moxibustion Medicine, College of Korean Medicine, Gachon University)
,
Song, Ho-Sueb
(Department of Acupuncture & Moxibustion Medicine, College of Korean Medicine, Gachon University)
Background: This study investigated the effects of Vipera lebetina turanica snake venom (SV) on cerebral infarction induced by middle cerebral artery occlusion in mice. Methods: Following cerebral infarction, SV was injected intravenously or added to BV2 cell culture. Tissue injury was detected usin...
Background: This study investigated the effects of Vipera lebetina turanica snake venom (SV) on cerebral infarction induced by middle cerebral artery occlusion in mice. Methods: Following cerebral infarction, SV was injected intravenously or added to BV2 cell culture. Tissue injury was detected using triphenyltetrazolium chloride (TTC) staining, neurological deficit score, NO, ROS, and GSH/GSSG assays, qPCR, Western blot, and cell viability. Results: Cerebral infarction caused by middle cerebral artery occlusion as observed by TTC staining, showed SV inhibited cell death, reducing the number of brain cells injured due to infarction. SV treatment for cerebral infarction showed a significant decrease in abnormal behavior, as determined by the neurological deficit score. The oxidation and inflammation of the cells that had cerebral infarction caused by middle cerebral artery occlusion (NO assay, ROS, GSH/GSSG assay, and qPCR), showed significant protection by SV. Western blot of brain infarction cells showed the expression of iNOS, COX-2, p-IkB-${\alpha}$, P38, p-JNK, p-ERK to be lower in the SV group. In addition, the expression of IkB increased. BV2 cells were viable when treated with SV at $20{\mu}g/mL$ or less. Western blot of BV2 cells, treated with 0.625, 1.5, $2.5{\mu}g/mL$ of SV, showed a significant decrease in the expression of p-IkB-${\alpha}$, p-JNK, iNOS, and COX-2 on BV2 cells induced by LPS. Conclusion: SV showed anti-inflammatory and anti-oxidant effects against cerebral infarction and inflammation.
Background: This study investigated the effects of Vipera lebetina turanica snake venom (SV) on cerebral infarction induced by middle cerebral artery occlusion in mice. Methods: Following cerebral infarction, SV was injected intravenously or added to BV2 cell culture. Tissue injury was detected using triphenyltetrazolium chloride (TTC) staining, neurological deficit score, NO, ROS, and GSH/GSSG assays, qPCR, Western blot, and cell viability. Results: Cerebral infarction caused by middle cerebral artery occlusion as observed by TTC staining, showed SV inhibited cell death, reducing the number of brain cells injured due to infarction. SV treatment for cerebral infarction showed a significant decrease in abnormal behavior, as determined by the neurological deficit score. The oxidation and inflammation of the cells that had cerebral infarction caused by middle cerebral artery occlusion (NO assay, ROS, GSH/GSSG assay, and qPCR), showed significant protection by SV. Western blot of brain infarction cells showed the expression of iNOS, COX-2, p-IkB-${\alpha}$, P38, p-JNK, p-ERK to be lower in the SV group. In addition, the expression of IkB increased. BV2 cells were viable when treated with SV at $20{\mu}g/mL$ or less. Western blot of BV2 cells, treated with 0.625, 1.5, $2.5{\mu}g/mL$ of SV, showed a significant decrease in the expression of p-IkB-${\alpha}$, p-JNK, iNOS, and COX-2 on BV2 cells induced by LPS. Conclusion: SV showed anti-inflammatory and anti-oxidant effects against cerebral infarction and inflammation.
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제안 방법
The inhibitory effect of SV in the presence of LPS (1 µg/mL) on cell growth of human keratinocyte BV2 cells was investigated. Cell viability by MTT assay and cell count was analyzed. The cells were treated with a concentration range of SV (0, 0.
SV maintains IκB levels which prevents NF-κB activation and the spread of iNOS and COX2, ultimately eliminating the spread of inflammation. During inflammation, tissues are oxidized, as seen in the ROS and GSH/ GSSG assays in this study. SV treatment prevented oxidation and, as seen in qPCR, reduced the level of inflammatory cytokines.
The cells were treated with a concentration range of SV (0, 0.313, 0.625, 1.25, 2.5, 5, 10, 20, 40 µg/mL) for 24 hours in the presence of LPS.
대상 데이터
at 37℃. The medium used was DMEM, supplemented with 10% FBS, and penicillin (100 U/mL). Equal amounts of total protein (40 μg) isolated from Microglial BV-2 cells were resolved on 8% or 10% sodium dodecyl sulfate polyacrylamide gels and then transferred to nitrocellulose membranes (Hybond ECL; Amersham Pharmacia Biotech, Piscataway, NJ, USA).
데이터처리
Differences in the data were assessed by a 1-way analysis of variance and when p = 0.05, the differences were assessed by Dunnett’s test.
성능/효과
After that, measuring NO and ROS, it indicates statistically significant differences from SV treated group. (B) After treatment of SV (50 ng/kg), ROS were decreased, but NO was not significantly different from MCAO group. ROS convergence has been reduced by the group that has handled SV in MCAO.
In conclusion, in the present study, SV was found to repress cerebral ischemia regardless of preexisting conditions or complications, and all animals treated with SV showed a reduction in cerebral ischemia. The main treatment target after cerebral ischemia, is not necrosis of the core, but rather apoptosis of the penumbral region.
On the day of implementation, neither the experimental nor the control group showed significant changes in neurological deficit score, but after 24 hours, the SV treatment group had a neurological deficit score significantly lower than on the implementation day (p < 0.05; Fig. 2).
The main treatment target after cerebral ischemia, is not necrosis of the core, but rather apoptosis of the penumbral region. The findings in this study indicate that SV effectively ameliorated neuron damage in the penumbral region, and markedly reduced ischemic damage in ICR mice. In view of the reduced levels of iNOS and COX-2, SV might attenuate infarct areas via an anti-inflammatory cytokine related effect through the reduction in NF-κB activity.
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