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Investigation of anti-oxidative stress in vitro and water apparent diffusion coefficient in MRI on rat after spinal cord injury in vivo with Tithonia diversifolia ethanolic extracts treatment.

Juang CL, Yang FS, Hsieh MS, Tseng HY, Chen SC, Wen HC - BMC Complement Altern Med (2014)

Bottom Line: The objective of the current study is to evaluate the anti-oxidative effect of Tithonia diversifolia ethanolic extracts (TDE) on cells and apply the pharmacological effect to SCI model using a MRI imaging algorism.TDE treatment slightly decreased the ADC level after 1-week SCI compared with control animals.Our studies have proved that the cytoprotection effect of TDE, at least in part, is through scavenging ROS to eliminate intracellular oxidative stress and highlight a potential therapeutic consideration of TDE in alternative and complementary medicine.

View Article: PubMed Central - PubMed

Affiliation: Department of Optometry, Yuanpei University, Hsinchu 30015, Taiwan. sjwen@mail.ypu.edu.tw.

ABSTRACT

Background: Spinal cord injury (SCI)-induced secondary oxidative stress associates with a clinical complication and high mortality. Treatments to improve the neurological outcome of secondary injury are considered as important issues. The objective of the current study is to evaluate the anti-oxidative effect of Tithonia diversifolia ethanolic extracts (TDE) on cells and apply the pharmacological effect to SCI model using a MRI imaging algorism.

Methods: The anti-oxidation properties were tested in a 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay. Rat liver cells (clone-9) were treated with various doses of TDE (0 ~ 50 μg/ml) before exposed to 250 μM H2O2 and cell survival was determined by MTT and LDH assays. We performed water apparent diffusion coefficient (ADC) map in MR techniques to investigate the efficacy of TDE treatment on SCI animal model. We performed T5 laminectomy and compression (50 g, 1 min) to induce SCI. PHILIP 3.0 T MRI was used to image 24 male Sprague-Dawley rats weighing 280-320 g. Rats were randomly divided into three groups: sham group, SCI group, SCI treated with TDE group. The MRI images were taken and ADC were acquired before and after of treatment of TDE (50 mg/kg B. W. orally, 5 days) in SCI model.

Results: TDE protected clone-9 cells against H2O2-induced toxicity through DPPH scavenging mechanism. In addition, SCI induced the increase in ADC after 6 hours. TDE treatment slightly decreased the ADC level after 1-week SCI compared with control animals.

Conclusion: Our studies have proved that the cytoprotection effect of TDE, at least in part, is through scavenging ROS to eliminate intracellular oxidative stress and highlight a potential therapeutic consideration of TDE in alternative and complementary medicine.

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Related in: MedlinePlus

Cytoprotective effect of TDE on hydrogen peroxide induced toxicity in clone 9 cells by LDH assays. LDH released in medium was assayed in (A) and LDH leakaged from cytosol was assayed in (B) after triton X-100 treatment. **p < 0.01; ***p < 0.001 versus hydrogen peroxide-treated cells. + p < 0.01 versus vit C group. Control group refers to cell free medium.
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Fig3: Cytoprotective effect of TDE on hydrogen peroxide induced toxicity in clone 9 cells by LDH assays. LDH released in medium was assayed in (A) and LDH leakaged from cytosol was assayed in (B) after triton X-100 treatment. **p < 0.01; ***p < 0.001 versus hydrogen peroxide-treated cells. + p < 0.01 versus vit C group. Control group refers to cell free medium.

Mentions: We then tested the anti-oxidative effects of TDE in cell culture system. The clone 9 was pretreated with TDE of different doses (0-50 μg/ml), followed by exposing the cells in 250 μM H2O2. Cell survival was detected by MTT assay and LDH release assay. Vitamin C treatment was used as a positive control (Figure 2). We found that treatment of H2O2 alone resulted in significant decrease of cell viability. Pretreatment of TDE protected H2O2-induced decrease of cell viability (Figure 2). Similar results were found in LDH release assay, where H2O2-induced release of LDH into medium from cytosol was significantly reduced by TDE protection (Figure 3A) and cytosolic LDH was significantly higher with TDE treatment than without TDE treatment (Figure 3B). These results suggest that TDE possesses anti-oxidative effect against H2O2-induced cytotoxicity.Figure 2


Investigation of anti-oxidative stress in vitro and water apparent diffusion coefficient in MRI on rat after spinal cord injury in vivo with Tithonia diversifolia ethanolic extracts treatment.

Juang CL, Yang FS, Hsieh MS, Tseng HY, Chen SC, Wen HC - BMC Complement Altern Med (2014)

Cytoprotective effect of TDE on hydrogen peroxide induced toxicity in clone 9 cells by LDH assays. LDH released in medium was assayed in (A) and LDH leakaged from cytosol was assayed in (B) after triton X-100 treatment. **p < 0.01; ***p < 0.001 versus hydrogen peroxide-treated cells. + p < 0.01 versus vit C group. Control group refers to cell free medium.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4289305&req=5

Fig3: Cytoprotective effect of TDE on hydrogen peroxide induced toxicity in clone 9 cells by LDH assays. LDH released in medium was assayed in (A) and LDH leakaged from cytosol was assayed in (B) after triton X-100 treatment. **p < 0.01; ***p < 0.001 versus hydrogen peroxide-treated cells. + p < 0.01 versus vit C group. Control group refers to cell free medium.
Mentions: We then tested the anti-oxidative effects of TDE in cell culture system. The clone 9 was pretreated with TDE of different doses (0-50 μg/ml), followed by exposing the cells in 250 μM H2O2. Cell survival was detected by MTT assay and LDH release assay. Vitamin C treatment was used as a positive control (Figure 2). We found that treatment of H2O2 alone resulted in significant decrease of cell viability. Pretreatment of TDE protected H2O2-induced decrease of cell viability (Figure 2). Similar results were found in LDH release assay, where H2O2-induced release of LDH into medium from cytosol was significantly reduced by TDE protection (Figure 3A) and cytosolic LDH was significantly higher with TDE treatment than without TDE treatment (Figure 3B). These results suggest that TDE possesses anti-oxidative effect against H2O2-induced cytotoxicity.Figure 2

Bottom Line: The objective of the current study is to evaluate the anti-oxidative effect of Tithonia diversifolia ethanolic extracts (TDE) on cells and apply the pharmacological effect to SCI model using a MRI imaging algorism.TDE treatment slightly decreased the ADC level after 1-week SCI compared with control animals.Our studies have proved that the cytoprotection effect of TDE, at least in part, is through scavenging ROS to eliminate intracellular oxidative stress and highlight a potential therapeutic consideration of TDE in alternative and complementary medicine.

View Article: PubMed Central - PubMed

Affiliation: Department of Optometry, Yuanpei University, Hsinchu 30015, Taiwan. sjwen@mail.ypu.edu.tw.

ABSTRACT

Background: Spinal cord injury (SCI)-induced secondary oxidative stress associates with a clinical complication and high mortality. Treatments to improve the neurological outcome of secondary injury are considered as important issues. The objective of the current study is to evaluate the anti-oxidative effect of Tithonia diversifolia ethanolic extracts (TDE) on cells and apply the pharmacological effect to SCI model using a MRI imaging algorism.

Methods: The anti-oxidation properties were tested in a 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay. Rat liver cells (clone-9) were treated with various doses of TDE (0 ~ 50 μg/ml) before exposed to 250 μM H2O2 and cell survival was determined by MTT and LDH assays. We performed water apparent diffusion coefficient (ADC) map in MR techniques to investigate the efficacy of TDE treatment on SCI animal model. We performed T5 laminectomy and compression (50 g, 1 min) to induce SCI. PHILIP 3.0 T MRI was used to image 24 male Sprague-Dawley rats weighing 280-320 g. Rats were randomly divided into three groups: sham group, SCI group, SCI treated with TDE group. The MRI images were taken and ADC were acquired before and after of treatment of TDE (50 mg/kg B. W. orally, 5 days) in SCI model.

Results: TDE protected clone-9 cells against H2O2-induced toxicity through DPPH scavenging mechanism. In addition, SCI induced the increase in ADC after 6 hours. TDE treatment slightly decreased the ADC level after 1-week SCI compared with control animals.

Conclusion: Our studies have proved that the cytoprotection effect of TDE, at least in part, is through scavenging ROS to eliminate intracellular oxidative stress and highlight a potential therapeutic consideration of TDE in alternative and complementary medicine.

Show MeSH
Related in: MedlinePlus