目的 驗(yàn)證炎調(diào)方對膿毒癥急性肺損傷（ALI）大鼠的保護(hù)效應(yīng)，并觀察炎調(diào)方對核因子κB（NF-κB）信號通路主要指標(biāo)活性調(diào)控的時效關(guān)系。方法 清潔級健康雄性SD大鼠隨機(jī)分為假手術(shù)組、模型組、炎調(diào)方（生藥量為9.9 g/kg）組、地塞米松（0.45 mg/kg）組，均每天ig給藥1次，連續(xù)給藥3 d。假手術(shù)組、模型組予等體積生理鹽水，末次給藥2 h后進(jìn)行手術(shù)。采用盲腸結(jié)扎穿孔術(shù)（CLP）制備膿毒癥ALI模型，各組分別于造模后4、6、8、10、12、18、24 h進(jìn)行HE染色后肺組織損傷程度評分、檢測肺組織NF-κB/p65 mRNA表達(dá)量和血清NF-κB、腫瘤壞死因子（TNF-α）、白細(xì)胞介素（IL）-1β、IL-6、IL-8水平。結(jié)果 與模型組比較，炎調(diào)方組造模后12、18、24 h肺組織損傷程度評分均顯著降低（P<0.01）。炎調(diào)方組和地塞米松組肺組織NF-κB/p65 mRNA相對表達(dá)量、血清NF-κB水平均顯著低于模型組（P<0.01）；高于假手術(shù)組（P<0.01）；炎調(diào)方組大鼠肺組織NF-κB/p65 mRNA相對表達(dá)量、血清NF-κB均隨CLP后時間的延長呈現(xiàn)上升趨勢，12 h后變化趨緩。不同時間點(diǎn)炎調(diào)方組和地塞米松組血清TNF-α、IL-1β、IL-6、IL-8均顯著低于模型組（P<0.01），顯著高于假手術(shù)組（P<0.01）；炎調(diào)方組血清TNF-α、IL-1β、IL-6、IL-8隨CLP后時間的延長呈現(xiàn)上升趨勢。膿毒癥ALI大鼠肺組織NF-κB/p65mRNA相對表達(dá)量與血清NF-κB和促炎因子TNF-α、IL-1β、IL-6、IL-8水平呈強(qiáng)正相關(guān)。結(jié)論 炎調(diào)方對膿毒癥ALI大鼠肺組織保護(hù)效應(yīng)的機(jī)制與下調(diào)NF-κB基因表達(dá)、進(jìn)而下調(diào)炎性信號通路下游的促炎細(xì)胞因子水平相關(guān)。

Objective To verify the effect of Yantiao-Prescription protectingts on rats from sepsis-induced acute lung injury, and to investigate the time-effect relationship on the main observation indexes activity in NF-κB signaling pathway. Methods Healthy male SD rats of clean grade were randomly divided into sham operation group, model group, Yantiao-Prescription (crude drug dosage:9.9 g/kg) group and dexamethasone (0.45 mg/kg) group. All rats were ig once a day for three consecutive days. The sham operation group and model group were given equal volume of normal saline, and the operation was performed 2 h after the last administration. Acute lung injury model induced by sepsis was established in rats by CLP. At 4, 6, 8, 10, 12, 18 and 24 h after CLP, 10 rats of each group were sacrificed and lung tissue and serum samples were collected. The pathological changes of lung tissue were observed by HE staining and by grading injury degree. The expression of NF-κB/p65 mRNA in lung tissue was detected by real-time PCR. The levels of NF-κB, TNF-α, IL-1β, IL-6 and IL-8 in serum were detected by ELISA. Results Compared with the model group, the scores of lung tissue injury in the Yantiao-prescription group were significantly lower than those in the model group (P<0.01). The relative expression of NF-κ B/p65 mRNA in lung tissue and serum NF-κB level in both groups were significantly lower than those in model group (P<0.01), and higher than that in sham operation group (P<0.01). The relative expression of NF-κB/p65 mRNA in lung tissue and serum NF-κB in Yantiao-Prescription group increased with the prolongation of CLP, and the change was slowed down after 12 h. At different time points, serum TNF-α, IL-1β, IL-6, IL-8 in Yantiao-Prescription group and dexamethasone group were significantly lower than those in model group (P<0.01), and significantly higher than those in sham operation group (P<0.01); serum TNF-α, IL-1β, IL-6, IL-8 in Yantiao-Prescription group increased with the time after CLP. There was a strong positive correlation between the relative expression of NF-κB/p65 mRNA and serum levels of NF-κB and proinflammatory factors TNF-α, IL-1β, IL-6 and IL-8. Conclusions Yantiao-Prescription protects the rats from sepsis-induced acute lung injury by depressing the expression of NF-κB gene, and then by down regulating the proinflammatory cytokines activity in the downstream of inflammatory signaling pathway.

R285.5

國家自然科學(xué)基金資助項(xiàng)目（81303105）