| Purpose and background:Subarachnoid hemorrhage (SAH) is a common clinical disease with high mortality and morbidity, accounting for about5‐7%of all stroke cases. The research usually focused on the cerebral vasospasm and treatment after SAH in the past. However, studies show that the treatment depend only on the control of vasospasm does not particularly effective in improving the prognosis of SAH. The research of early brain injury after SAH increased gradually recently, whether having benefits to reduce the incidence and improve the prognosis through control the early brain injury after SAH is become an important research direction. Butylphthalide extracts from the seed of celery firstly, which is now a kind of synthetic racemic, mainly used for the treatment of acute ischemic stroke. Present studies show that it have the effect of improving cerebral blood flow, mitigating cerebral edema, protecting never cell, blocking the injury caused by ischemic, improving the metabolism of the brain, inhibiting thrombosis and reducing neuronal apoptosis. The pathological process after SAH is similar to ischemic stroke after SAH occurs, and there is rare report applying this medicine after SAH occurs, Whether have protecting effect during pathological process of SAH, improving the prognosis is not yet clear. Thus, this experiment will apply Butylphthalide on SAH rat model, hope to find out whether there is a protective effect to the early brain injury in a rat SAH model through detection MDA, TNF‐α, IL‐1β, ICAM‐1and cell death of the brain organization.Materials and Methods:1. The experimental materialAdult SPF SD rats(N=60),30female and30male, weighing about250~330g, provided by the Experimental Animal Center of Guangdong Province, fed in the SPF animals room of Guangzhou Medical University Experimental Animal Center with normal fodder. About1~2weeks later establish the model of subarachnoid hemorrhage with the method which called magna cisterna blood injection.2. The experimental methodDivide the animal into6groups, the sham group, the control group and the experimental group. Each of the group has two subset, a24h group and a12h group, thus each group contains10animals.5animals in each group were taken for tissue, and the remaining5for slice. After building the SAH model, Butylphthalide is injected to the animals of experimental group (2.5mg/kg), the same volume of normal saline replaced the Butylphthalide for the control group. After get the homogenate of rat brain, measuring the content of MDA with the method of thiobarbituric acid (TBA), and detected the content of necrosis factor α (TNF‐α), interleukin rats1β (IL‐1β), rat cell adhesion molecule1(ICAM‐1) levels with the method of double‐antibody sandwich assay (ELISA). The total protein content also have to be measured with the method of bicinchoninic acid (BCA) as a standard for other target. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) is used to detect the cell death, after getting the images of each slices, use the Image‐Pro Plus6.0program to count the death cells and all other normal cells. The result is analysis with t‐test and χ2test. When P <0.05was considered there is statistically significant.Result1. MortalityThere is no death in Sham group, and between the12h control group and experimental group,the24h control group and experimental group there is no different(p>0.05).2. MDAThere is no significant between the12h sham group and24h sham group (P>0.05). Comparing to the sham group, the content of MDA of all other groups is elevated (P<0.05). The12h experimental group is a little lower the24h experimental group, the same condition is also between the12h control group and24h control group, but there is no significant difference (P>0.05). The MDA content of12h experimental group is lower than the12h control group (p<0.05), also the24h groups.3. TNF‐α,IL‐β and ICAM‐1There is no significant between the12h sham group and24h sham group (P>0.05). Comparing to the sham group, the content of TNF‐α of all other groups is elevated (P<0.05). The12h experimental group is a little lower the24h experimental group, the same condition is also between the12h control group and24h control group, but there is no significant difference (P>0.05). The TNF‐α content of12h experimental group is lower than the12h control group (p<0.05), also the24h groups. The contents of IL‐β and ICAM‐1‐is very similar to the tnf‐α, the experimental group have a lower content of those cytokines than the control group. The sham group has extreme low content of those substances.4. Cell deathThere is rare death cells in the12h and24h sham group, the death cell of the12h control group is higher than the12h experimental group (p<0.05), the death cell of the24h control group is also higher than the24h experimental group (p<0.05). The24h control group have more death cell then the12h control group without significant difference (p>0.05), the24h experimental group also have a higher content of death cells than the12experimental group (p<0.05).ConclusionEarly after SAH, the content of MDA, TNF‐α, IL‐β, and ICAM‐1in the brain tissue were increased, which may be related to the decrease of blood flow, the release of hemoglobin and other substances into the subarachnoid space after SAH. Butylphthalide can reduce the expression of inflammatory molecules, the occurrence of inflammatory response and cell death after SAH. It have a protective effect to the nerve injury after SAH. |
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