益气活血法通过抑制细胞焦亡减轻缺血性脑中风急性期大鼠脑组织炎症

Yiqi Huoxue Therapy Alleviates Cerebral Inflammation in Rats with Acute Ischemic Stroke by Inhibiting Pyrophosis

【目的】探讨益气活血法对缺血性脑中风急性期大鼠脑组织炎症的作用及机制。【方法】将47只SD大鼠留取9只作为假手术组,其余大鼠建造缺血性脑中风急性期模型。将造模成功的30只大鼠随机分为中药低、高剂量组及模型组,每组10只。中药低、高剂量组灌胃益气活血方汤药4.4、8.8 g/kg,模型组、假手术组灌胃等体积生理盐水,每天1次,连续给药15 d。给药第3、5、7天,测定大鼠神经功能变化;给药结束后,以2,3,5-氯化三苯基四氮唑(TTC)染色观察大鼠脑梗死情况,酶联免疫吸附分析(ELISA)检测大鼠脑组织匀浆中白细胞介素(IL)-1β、肿瘤坏死因子α(TNF-α)、IL-18含量,苏木素-伊红(HE)染色观察大鼠脑组织病理学表现,蛋白免疫印迹(Western Blot)法检测大鼠脑缺血半暗带组织嘌呤能受体P2X7(P2RX7)、半胱氨酸天冬氨酸蛋白酶(Caspase)-1、Caspase-11、消皮素D(GSDMD)表达水平。【结果】与假手术组比较,模型组大鼠神经功能评分、脑梗死率,脑组织IL-1β、TNF-α、IL-18水平,脑缺血半暗带组织P2RX7、Caspase-1、Caspase-11、GSDMD蛋白相对表达量升高(P<0.05);与模型组比较,中药低、高剂量组大鼠神经功能评分、脑梗死率,脑组织IL-1β、TNF-α、IL-18水平,脑缺血半暗带组织P2RX7、Caspase-1、Caspase-11、GSDMD蛋白相对表达量降低(P<0.05)。模型组大鼠脑组织大面积炎症渗出,水肿明显,细胞崩解,神经元空泡变性或呈点状坏死,而中药低、高剂量组脑组织炎症渗出、水肿明显减轻,细胞形态、神经元空泡变性改善。【结论】益气活血法可通过抑制细胞焦亡,减轻缺血性脑中风急性期大鼠脑组织炎症。

Objective To explore the effect of Yiqi Huoxue therapy on cerebral inflammation in rats with acute ischemic stroke and mechanism. Methods Nine out of 47 SD rats were selected as sham-operation group,and the others were constructed as acute ischemic stroke model. Thirty successful modeled rats were randomly divided into the low-dose,high-dose Chinese herbal medicine groups and model group,with 10 rats in each group. The lowdose,high-dose Chinese herbal medicine groups were intragastrically given Yiqi Huoxue Decoction at 4.4,8.8 g/kg,respectively,and the model group and sham-operation group were intragastrically given equal volume of normal saline, once a day, both administrations lasting for consecutive 15 days. On day 3, 5, 7 of treatment, the changes of nerve function in rats were measured. After treatment, 2,3,5-triphenyltetrazole chloride(TTC)staining was used to observe the cerebral infarction in rats,the contents of interleukin(IL)-1β,tumor necrosis factor α(TNF-α) and IL-18 in brain tissue homogenates of rats were determined by enzyma-linked immunosorbent assay(ELISA), hematoxylin-eosin(HE)staining was used to observe the histopathologyical features of rat brain tissue, the expression levels of purinergic receptor P2 X7(P2 RX7), cysteine aspartatespecific protease(Caspase)-1, Caspase-11 and pyloderma D(GSDMD)in ischemic penumbra of rats were detected by Western Blot. Results Compared with the sham-operation group, the neurological function score,cerebral infarction rate,levels of IL-1β,TNF-α and IL-18 in brain tissue,and the relative protein expression levels of P2 RX7,Caspase-1,Caspase-11 and GSDMD in cerebral ischemic penumbra tissue were increased in the model group(P<0.05);compared with the model group,the neurological function score,cerebral infarction rate,levels of IL-1β,TNF-α and IL-18 in brain tissue,and the relative protein expression levels of P2 RX7,Caspase-1,Caspase-11 and GSDMD in cerebral ischemic penumbra tissue were decreased in low-dose and highdose Chinese herbal medicine groups(P<0.05). In the model group,large area of inflammation exudate,marked edema,cell disintegration,vacuolar degeneration or spotty necrosis of neurons were seen,but the inflammatory exudation and edema of brain tissue were significantly reduced in the low-dose and high-dose Chinese herbal medicine groups, and the cell morphological manifestation and neuron vacuolar degeneration were improved.Conclusion Yiqi Huoxue therapy is effective for reducing the cerebral inflammation in rats with acute ischemic stroke by inhibiting pyrophosis.