化痰通络汤改善大鼠脑缺血再灌注后继发肺炎性损伤作用的研究

Protective effect of HuatanTongluo Decoction on lung inflammatory injury secondary to cerebral ischemia-reperfusion in rats

目的 探讨化痰通络汤(HuatanTongluo decoction, HTD)对大鼠脑缺血再灌注后继发肺组织炎性损伤的改善作用。方法 48只SPF级Sprague–Dawley大鼠随机分为假手术组、模型组、HTD低、中、高剂量组和阳性对照药物(依达拉奉+丁酸梭菌)组。采用阻塞右侧大脑中动脉建立脑缺血再灌注(Middle cerebral artery occlusion reperfusion, MACO/R)模型,模型制备前2h给药,HTD和丁酸梭菌灌胃给药,依达拉奉腹腔注射给药。肺组织炎性损伤用HE染色法评估;采用qRT-PCR法和ELISA法检测肺组织炎症因子的转录水平和含量;WB法检测Sirt1、p38MAPK、p-p38MAPK的蛋白水平。结果 MACO/R导致肺组织炎性损伤,肺损伤评分显著增加,且促炎因子IL-1β、IL-6和TNF-α含量显著升高,而抑炎因子IL-10含量显著降低。中、高剂量HTD和阳性药物可显著降低肺损伤评分以及IL-1β、IL-6和TNF-α含量,增加IL-10含量,且逆转脑缺血再灌注后诱导的肺组织p38MAPK磷酸化水平的升高和Sirt1蛋白水平的下调。结论 HTD可能通过上调Sirt1表达和下调p38MAPK的磷酸化,抑制IL-1β、IL-6、TNF-α和促进IL-10在肺组织的表达,进而发挥对MACO/R后继发肺炎性损伤的改善作用。

Objective To explore the protective effect of HuatanTongluo Decoction(HTD) on lung inflammatory injury secondary to middle cerebral artery ischemia-reperfusion(MACO/R) in rats. Methods 48 SPF rats(male) were randomly divided into sham operation group, cerebral ischemia-reperfusion model group, low/medium/high dose group of HTD, and positive group(edaravone+Clostridium butyricum). The model was established by unilateral occlusion of the middle cerebral artery. HTD and Clostridium butyricum were administered by ig 2h before model preparation, and edaravone was injected intraperitoneally. Inflammatory injury of rat lung was evaluated by hematoxylin eosin staining(HE). Quantitative real-time polymerase chain reaction(qRT-PCR) and enzyme linked immunosorbent assay(ELISA) were used to assess the mRNA level and secretion of inflammatory cytokines(IL-1β, IL-6, IL-10, and TNF-α) in rat lung. The protein level of p38MAPK, p-p38MAPK, and Sirt1 was detected by Western blotting(WB).Results MACO/R caused inflammation in rat lung, an increase of score of lung injury, the elevated levels of pro-inflammatory cytokines(IL-1β, IL-6, and TNF-α) and the reduced level of anti-inflammatory cytokine(IL-10). Compared with model group, the scores of lung injury in medium/high dose group and positive group were significantly decreased. The results of qRT-PCR and ELISA showed that the medium/high dose of HTD significantly inhibited the increase in the transcription and release of IL-1β, IL-6, and TNF-α as well as the decrease in IL-10 transcription and release induced by MACO/R in rat lung. In addition, the medium/high dose of HTD reversed the upregulation of p38MAPK phosphorylation, and the downregulation of Sirt1 protein, secondary to MACO/R middle cerebral artery ischemia-reperfusion. Conclusion HTD may reduce the levels of IL-1β, IL-6, and TNF-α and raise the level of IL-10 by upregulating Sirt1 expression and downregulating p38MAPK phosphorylation, to protective against the rat lung injury secondary to MACO/R.