| BackgroundCerebrovascular Diseases（CVD）is the most common and multiple neurologicaldisease. In recent years，it has become the leading cause of death and disabilityof the global population, and a serious threat to human health and safety. Whileacute ischemic stroke (cerebral infarction) accounting for about60%to80%ofall cases is the most common type of stroke. Even in the survival of patients withstroke, still there are varying degrees of neurological dysfunction, lead to varyingdegrees of disability, carry a heavy burden to families and society.Salvia is a kind of dicotyledons Labiatae plant. It has been used for more thantwo thousand years in traditional Chinese medicine. According to the theory oftraditional Chinese medicine, Salvia has the effect on blood to regulatemenstruation, stasis and relieve pain, consumption carbuncle, soothe the nerves.Tanshinone Ⅱ A is diterpenoid quinone compound isolated from Salvia, themain active ingredient of Salvia. Tanshinone Ⅱ A sulfonate (DS201) iswater-soluble substances by sulfonation, made from the fat-soluble tanshinone IIA.It has been around for more than thirty years as a traditional Chinese medicinemonomer formulation of the active ingredient. Widely used in the treatment ofheart and lung diseases, and types of skin and superficial tissue inflammation.Studies shown that tanshinone improving coronary collateral circulation, reducing the area of the infarcted myocardium, while lifting micro vascularspasm, increase the flow rate and the flow of the microcirculation, eliminate localvenous stasis, improve tissue cells metabolic disorders caused by ischemia andhypoxia, inhibit lung fibrosis, lung disease and reduce lung tissue inflammatorycell infiltration, increased pulmonary capillary blood flow.Recent years, based on a large number of studies, the applications oftanshinone II A in the field of cerebrovascular disease have been paid graduallyattention. This experiment is intended to study the protective effect on cerebralischemia-reperfusion injury.ObjectiveAnalysis the cerebral infarction volume, brain water content, and neurologicalscore at different time points of the rats in each group, to study the protectiveeffects of tanshinone Ⅱ A sulfonate on focal cerebral ischemia-reperfusioninjury model, and explore its possible mechanism.Methods1. Experimental animals and groups:40SD rats, weight202-305(247.57±32.09) g. All rats were randomly divided into four groups, for the shamgroup, model of focal cerebral ischemia group (abbreviation: the modelgroup), tanshinone Ⅱ A sulfonate pretreatment group (abbreviation:pretreatment group), tanshinone Ⅱ A sulfonate treatmentgroup(abbreviation: the treatment group),10in each group.2. Model Preparation: Fabricated models (Middle cerebral artery occlusion,MCAO) by modified Longa [9] suture, opening up the blood flow two hoursafter surgery, lead to the formation of reperfusion.3. The method of administration: Rats of sham operation group and modelgroup were given intraperitoneal injection of saline2ml,30minutes before modeling surgery; rats of pretreatment group were given intraperitonealinjection of tanshinone Ⅱ A sulfonate30mg/kg,30minutes beforemodeling surgery; rats of treatment group were given intraperitonealinjection of sodium tanshinone Ⅱ A30mg/kg while opening up the bloodflow.4. Cerebral Infarction Volume: Rats were killed at24h after reperfusion, thebrain tissue is cut into thin slices, stained by TTC (2,3,5–triphenyltetrazolium chloride). Take digital pictures, using computer imageanalysis software (HPIAS-1000) to calculate the cerebral infarction volume.5. Brain Water Content: After model rats were killed, remove the completebrain. Left infarcted hemisphere, measuring the wet weight with electronicprecision scales, and then placed into100℃electric oven to be constantweight, measured dry weight. Calculate the brain water content.6. Neurological Function Score: Application Zea Longa neurological functionscore at12hours and24hours after reperfusion, neurological scorerespectively. Evaluate neurological deficit situation of model rats. Scoringcriteria: no symptoms of nerve damage score0; inextensible contralateralanterior claw score1; the contralateral rotation score2; dumping to theopposite side score3; no independent activity associated with disturbance ofconsciousness score4; death score5. The higher score, the more seriousneurological impairment.7. TNF-α: Quantitative analysis of TNF-α in rats’ blood by Enzyme-linkedimmunosorbent assay.8. Statistical analysis: SPSS13.0statistical software for statistical analysis ofthe experimental data. Experimental data (), multi-group comparedusing one-way ANOVA, between the two groups using two-sample t test, P<0.05for the difference was statistically significant. Results1. After TTC staining, sham group has no obvious white infarcted area, theother groups showed visible obvious white infarcted areas. Statisticalanalysis, F=7.29, P <0.01, significant difference.In model group, thepercentage of infarct volume is higher than the pretreatment and treatmentgroups, t=13.70,11.26, P <0.01.2. Brain water content was significantly different. F=8.19，P＜0.01. Furtherpairwise comparisons show brain water content in model group was higherthan in pretreatment and treatment groups, t=3.13,2.97, P<0.05, significantdifference.3.12hours after surgery, each group neurological score comparison, F=4.72,P <0.05; after24hours, F=9.72, P <0.01. Showing that at different timepoints, the neurological score differences were significant. Further pairwisecomparisons show the score in model group higher than in the pre-treatmentgroup and treatment group, t=2.74,2.82,3.34,2.86, P <0.05, significantdifference.4. TNF-α content in each group was significantly different, F=3.96, P＜0.05.Further pairwise comparisons show model group is higher than that inpretreatment and treatment groups (t=2.78、2.12, P＜0.05）.ConclusionTanshinone Ⅱ A sulfonate can reduce ischemia-reperfusion rat model ofcerebral infarct volume and to reduce brain water content, and improveneurological function score. Inhibition of inflammatory cytokine release may beone of the mechanisms. |
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