丁苯酞通过抑制JNK/p38 MAPK信号通路减少脑梗死大鼠神经细胞的凋亡

To investigate the effects of butylphthalide on reducing neuronal apoptosis in rats with cerebral infarction by inhibiting the JNK/p38 MAPK signaling pathway

目的:探讨丁苯酞通过抑制JNK/p38 MAPK信号通路减少脑梗死大鼠神经细胞的凋亡。方法:将48只SD雄性大鼠分为DZ组(对照组)、CI组(模型组)与NBP组(丁苯酞组),CI组和NBP组大鼠建立脑梗死模型,NBP组采用NBP溶液80 mg·kg^-1·d^-1灌胃,剩余两组采用同体积的花生油灌胃,连续14 d处理后,采用Zea Longa评分对DZ组、CI组及NBP组的神经功能进行评分,TTC染色法观察DZ组、CI组和NBP组脑梗死体积,HE染色法观察DZ组、CI组和NBP组脑组织病理形态,TUNEL法检测DZ组、CI组和NBP组脑组织神经细胞凋亡情况,Western blot检测DZ组、CI组和NBP组脑组织p-JNK和p-p38MAPK的蛋白表达。结果:CI组神经功能高于DZ组,差异具有统计学意义(P<0.05);NBP组神经功能评分减轻与CI组相比,差异具有统计学意义(P<0.05);CI组脑梗死体积高于DZ组,差异具有统计学意义(P<0.05);NBP组出现轻微梗死体积少于CI组,差异具有统计学意义(P<0.05);DZ组神经细胞排序整齐,数量较多,CI组血管与脑间质之间空隙增大,细胞固缩严重,神经元结构不完整;NBP组与CI组相比神经元固缩减轻,数量增多;凋亡的神经细胞为褐色,CI组神经细胞的凋亡率高于DZ组,差异具有统计学意义(P<0.05);NBP组神经细胞凋亡率与CI组相比减少,差异具有统计学意义(P<0.05);Western blot结果显示:CI组p-JNK和p-p38MAPK蛋白表达高于DZ组,差异具有统计学意义(P<0.05),NBP组p-JNK和p-p38MAPK蛋白水平低于CI组,差异具有统计学意义(P<0.05)。结论:丁苯酞能够改善脑梗死大鼠神经功能损伤,减少凋亡神经细胞,降低梗死体积,与抑制JNK/p38 MAPK通路表达相关。

Objective To investigate the effects of butylphthalide on reducing neuronal apoptosis in rats with cerebral infarction by inhibiting the JNK/p38 MAPK signaling pathway.Methods Totally 48 SD male rats were divided into DZ group(control group),CI group(model group)and NBP group(butylphthalide group).Rats in CI group and NBP group were used to establish cerebral infarction models.NBP group used the solution of NBP(80 mg·kg^-1·d^-1)that was administered orally,and the remaining two groups were administered with the same volume of peanut oil.After 14 consecutive days of treatment,the Zea Longa scoring was used to evaluate the neurological function.TTC staining was used to observe the cerebral infarction volume of rats,and HE staining was used to observe the pathological morphology of brain tissue.TUNEL assay was used to detect the neuronal apoptosis,and Western blot was used to detect the expression of p-JNK and p-p38MAPK in brain tissues in all groups.Results The neurological function of the rats in the CI group was higher than that in the DZ group,and the difference was statistically significant(P<0.05).The neurological function score of the rats in the NBP group was reduced compared with the CI group,and the difference was statistically significant(P<0.05).The cerebral infarction volume in the group was 35.56%higher than that in the DZ group,and the difference was statistically significant(P<0.05).The minor infarct volume in the NBP group was 21.59%,which was less than that in the CI group,and the difference was statistically significant(P<0.05).Nerve cells were neatly displayed in a large number.The gap between blood vessels and interstitial tissue in the CI group was enlarged,the cells were severely contracted,and the neuron structure was incomplete.Compared with the CI group,the NBP group has reduced neuron contraction and increased number;the dead nerve cells were brown.The apoptosis rate of nerve cells in the CI group was 79.65%higher than that in the DZ group was 5.82%with statistically significant difference(P<0.05).The nerve cell apoptosis rate in the NBP group was 30.23%.Compared with CI group,the difference was statistically significant(P<0.05);Western blot results showed that p-JNK and p-p38MAPK protein expression in CI group was higher than that in DZ group,and the difference was statistically significant(P<0.05).The levels of p-JNK and p-p38MAPK proteins in the NBP group were lower than those in the CI group with statistically significant difference(P<0.05).Conclusion Butylphthalide can improve neurological damage,reduce apoptotic nerve cells,and reduce infarct volume in rats with cerebral infarction,which is related to the inhibition of JNK/P38 MAPK pathway expression.