复方丹参滴丸通过JAK-STAT通路对癫痫模型大鼠的保护机制研究

Protective Mechanism of Compound Danshen Dripping Pills on Epilepsy Model Rats Through JAK-STAT Pathway

目的:探讨复方丹参滴丸通过JAK-STAT通路对癫痫模型大鼠的保护机制。方法:建立癫痫大鼠模型,随机分为模型组、复方丹参滴丸组、托伐替尼(JAK抑制剂)组和复方丹参滴丸+托伐替尼组,每组12只;另取12只SD大鼠设为对照组。分组处理后,观察各组大鼠癫痫发作次数、持续时间;HE染色检测各组大鼠海马神经元病理变化;Morris水迷宫实验检测各组大鼠认知功能;酶联免疫吸附法检测各组大鼠血清肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)水平;免疫印记法检测各组大鼠脑组织JAK/STAT3通路相关蛋白表达。结果:与对照组相比,模型组大鼠海马神经元呈现排列紊乱、疏松且失去正常结构,胞体变为三角形或不规则图形,胞质浓缩、深染,胞核固缩,核膜、核仁不明显,胞浆呈空泡样改变,细胞凋亡,数量显著减少等病理损伤;癫痫发作次数明显增多,血清TNF-α、IL-6水平及脑组织p-JAK/JAK和p-STAT3/STAT3表达明显升高,持续时间、逃避潜伏期明显延长,穿越平台原位置次数明显减少,差异均有统计学意义(P<0.05)。与模型组相比,复方丹参滴丸组、托伐替尼组和复方丹参滴丸+托伐替尼组大鼠海马神经元病理损伤减轻,癫痫发作次数明显减少,血清TNF-α、IL-6水平及脑组织p-JAK/JAK和p-STAT3/STAT3表达明显降低,持续时间、逃避潜伏期明显缩短,穿越平台原位置次数明显增多,差异均有统计学意义(P<0.05);而复方丹参滴丸组大鼠逃避潜伏期与模型组的差异无统计学意义(P>0.05)。与复方丹参滴丸组、托伐替尼组相比,复方丹参滴丸+托伐替尼组大鼠海马神经元病理损伤进一步减轻,癫痫发作次数明显减少,血清TNF-α、IL-6水平及脑组织p-JAK/JAK和p-STAT3/STAT3表达明显降低,持续时间明显缩短,穿越平台原位置次数明显增多,差异均有统计学意义(P<0.05)。结论:复方丹参滴丸可改善癫痫模型大鼠症状,保护神经功能,可能通过抑制JAK/STAT3信号激活实现。

OBJECTIVE:To probe into the protective mechanism of compound Danshen dripping pills on epilepsy model rats through JAK-STAT pathway.METHODS:The rat model of epilepsy was established,and all rats were randomly divided into the model group,the compound Danshen dropping pill group,tofacitinib(JAK inhibitor)group and the compound Danshen dropping pill+tofacitinib group,with 12 rats in each group.Another 12 SD rats were selected as the control group.After grouping,the seizure frequency and duration were observed.HE staining was used to detect the pathological changes of hippocampal neurons in each group.Morris water maze test was used to detect the cognitive function of rats.The levels of serum tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)were measured by enzyme-linked immunosorbent assay(ELISA),and the expression of JAK/STAT3 pathway-related protein in brain tissue of rats in each group was detected by immunoblotting.RESULTS:Compared with the control group,the hippocampal neurons in the model group showed pathological injuries such as disordered arrangement,loosening and loss of normal structure,the cell body became triangular or irregular pattern,the cytoplasm was concentrated and deeply stained,the nucleus pyknosis,the nuclear membrane and nucleolus were not obvious,the cytoplasm showed vacuole-like changes,apoptosis and the number of cells decreased significantly.The seizure frequency,levels of serum TNF-αand IL-6,brain p-JAK/JAK and p-STAT3/STAT3 increased significantly,the duration and escape latency prolonged significantly,and the times of traversing the original platform position decreased significantly,with statistically significant difference(P<0.05).Compared with the model group,the pathological damage of hippocampal neurons in the compound Danshen dripping pill group,the tofacitinib group,the compound Danshen dripping pill+tofacitinib group reduced significantly,the seizure frequency,levels of serum TNF-αand IL-6,brain p-JAK/JAK and p-STAT3/STAT3 decreased significantly,the duration,escape latency shortened significantly,and the times of traversing the original platform position increased significantly,with statistically significant difference(P<0.05),while there was no significant difference in escape latency between compound Danshen dripping pill group and the model group(P>0.05).Compared with the compound Danshen dropping pill group and the tofacitinib group,the pathological damage of hippocampal neurons in the compound Danshen dropping pill+tofacitinib group was further alleviated,the seizure frequency,levels of serum TNF-αand IL-6,brain p-JAK/JAK and p-STAT3/STAT3 decreased significantly,the duration shortened significantly,and the times of traversing the original platform position increased significantly,with statistically significant difference(P<0.05).CONCLUSIONS:Compound Danshen dropping pill can improve the symptoms and protect the nerve function of epileptic model rats,which may be achieved by inhibiting the activation of JAK/STAT3 signal.