Abstract
BackgroundTo explore the neuroprotective effect and optimize the therapeutic dose and time window of picroside II by orthogonal test and the expression of myelin basic protein (MBP) in cerebral ischemic injury in rats. Bilateral common carotid artery occlusion (BCCAO) was used to establish forebrain ischemia models. The successful rat models were grouped according to orthogonal experimental design and injected picroside II intraperitoneally at different ischemic time with different doses. Myelin sheath fast green staining(FGS) and transmission electron microscopy (TEM) were used to observe nerve fiber myelin; the expression of MBP was tested qualitatively and quantitatively by immunohistochemical assay (IHC) and Western blot (WB); Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the transcription level of MBP mRNA.ResultsThe protective effect of picroside II was presented by increasing the expression of MBP and decreasing demyelination after cerebral ischemic injury. The best therapeutic time window and dose was (1) ischemia 2.0 h with picroside II 10 mg/kg body weight according to the results of FGS, IHC and WB; (2) ischemia 1.5 h with picroside II 20 mg/kg according to the analysis of RT-PCR.ConclusionGiven the principle of the longest time window and the lowest therapeutic dose, the optimized therapeutic dose and time window should be injecting picroside II intraperitoneally with 10-20 mg/kg body weight at ischemia 1.5-2.0 h in cerebral ischemic injury.
Highlights
To explore the neuroprotective effect and optimize the therapeutic dose and time window of picroside II by orthogonal test and the expression of myelin basic protein (MBP) in cerebral ischemic injury in rats
This indicated the therapeutic time window had significantly influence on the expression of myelin after cerebral ischemia injury, while no significant influence existed in different drug doses and time-dose interactions (Table 4)
Least significant differences (LSD) showed that different administration time between 1.0 h(A1) and 1.5 h(A2), 1.0 h(A1) and 2.0 h(A3), 1.5 h(A2) and 2.0 h (A3), 1.5 h(A2) and 2.5 h(A4), 2.0 h(A3) and 2.5 h(A4) had significant differences (P < 0.05), no significant differences found between the rest groups (P > 0.05)
Summary
To explore the neuroprotective effect and optimize the therapeutic dose and time window of picroside II by orthogonal test and the expression of myelin basic protein (MBP) in cerebral ischemic injury in rats. Animal experiments [6] proved that there was a small amount expression of MBP mRNA in the brain of normal adult rats, while the MBP mRNA [7] and protein [8] decreased time-dependently in early stage of cerebral limitation of neurobehavioral evaluation and immunohistochemical assay, we attempted to detect the expression levels of MBP in brain tissue qualitatively and quantitatively and observe the change of myelin structure through a various biological techniques, just to explore the optimal therapeutic dose and time window of picroside II after cerebral ischemic injury
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