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.

Chi-Long Juang,Su-Chiu Chen,Hsiao-Chuan Wen,Fei Shish Yang,Hu-Yi Tseng,Ming Shuang Hsieh

doi:10.1186/1472-6882-14-447

Abstract

BackgroundSpinal 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.MethodsThe 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.ResultsTDE 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.ConclusionOur 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.

Highlights

Spinal cord injury (SCI)-induced secondary oxidative stress associates with a clinical complication and high mortality
Free radical scavenging profile of Tithonia diversifolia ethanolic extracts (TDE) was expressed as % of DPPH inhibition compared to ascorbic acid
Similar results were found in lactate dehydrogenase (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)

Summary

Introduction

Spinal cord injury (SCI)-induced secondary oxidative stress associates with a clinical complication and high mortality. 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. Other than its anti-cancer activity, the sesquiterpenoid that extracted from other species shows its anti-inflammatory effect via inhibition of oxidative stress both in vitro and in vivo in the treatment of psoriasis [17]. Several research groups have performed clinical MR scanners for imaging small animal models. The goal of this study is seeking to successfully apply MRI for examining small rodents’ spinal cord injury using a standard clinical 3 T scanner and to keep instrumental cost low. While evaluating an acute ischemic stroke, ADC images may play a crucial role in identifying the degree of damage due to its higher water content in injured area

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