摘要
目的探讨羟基红花黄素A(HSYA)对脑卒中相关性肺炎(SAP)大鼠肺组织的保护作用及可能的机制。方法随机选取SD大鼠10只作为假手术组,其余大鼠采用四动脉阻断法制备SAP模型。将造模成功的SAP模型大鼠随机分为4组,分别用生理盐水(假手术组、SAP组)、Janus激酶2特异性抑制剂N-苄基-3,4-二羟基亚苄基氰基乙酰胺(AG490,5 mg/kg,AG490组)、低剂量(8 mg/kg,L-HSYA组)及高剂量HSYA(16 mg/kg,H-HSYA组)对大鼠进行治疗,连续给药7 d。用HE染色法观察各组肺组织病理变化;测量肺组织湿重量(W)、干重量(D),计算W/D值;用比色法检测肺组织中髓过氧化物酶(MPO)活性;用酶联免疫吸附法检测支气管肺泡灌洗液(BALF)中炎性细胞因子水平及肺组织核提取物中核因子κB(NF-κB)结合活性; Western blotting法检测肺组织核提取物中磷酸化Janus激酶2(p-JAK2)、磷酸化信号传导子及转录激活子3(p-STAT3)、核因子κB p65(NF-κB p65)蛋白表达。结果与假手术组比较,SAP组肺损伤评分、肺组织W/D值、MPO活性高,BALF中肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、IL-18水平及NF-κB DNA结合活性高,NF-κB p65、p-JAK2、p-STAT3蛋白表达高(P均<0. 05);与SAP组比较,AG490组、L-HSYA组及H-HSYA组肺损伤评分、肺组织W/D值、MPO活性低,BALF中TNF-α、IL-1β、IL-18水平及NF-κB DNA结合活性低,NF-κB p65、p-JAK2、p-STAT3蛋白表达低(P均<0. 05);且AG490组、HHSYA组以上指标变化最明显(P均<0. 05)。结论 HSYA能够减轻SAP大鼠肺组织的损伤及炎症反应,该作用可能与抑制肺组织中JAK2/STAT3信号通路激活有关。
Objective To investigate the protective effect of hydroxysafflor yellow A( HSYA) on lung tissues of stroke-associated pneumonia( SAP) rats and to explore its possible mechanism. Methods Ten SD rats were randomly selected as the sham operation group,and the other rats were prepared for the SAP models by 4-artery occlusion method. After modeling,the rats were treated with normal saline( sham group,SAP group),Janus kinase 2( JAK2) specific inhibitor N-benzyl-3,4-two hydroxybenzyl cyanoacetamide AG490( 5 mg/kg,AG490 group),low-dose HSYA( 8 mg/kg L-HSYA group) and high-dose HSYA( 16 mg/kg,H-HSYA group) for 7 consecutive days. Histopathological analysis was used to observe the lung injury. We measured the wet weight( W) and dry weight( D) of the lung tissue and calculated the W/D value. The activity of myeloperoxidase( MPO) in the lung tissues was detected by colorimetry. The level of inflammatory cytokines in bronchoalveolar lavage fluid( BALF) was detected by enzyme-linked immunosorbent assay. Enzyme linked immunosorbent assay was used to detect nuclear factor κB( NF-κB) binding activity in the lung tissues. Western blotting was used to analyze the expression of phosphorylated Janus kinase 2( p-JAK2),phosphorylated signal transducer and activator of transcription 3( p-STAT3),nuclear factor κB p65( NF-κB p65) in the nuclear extracts of the lung tissues. Results Compared with the sham group,the lung injury score,the W/D,MPO of the lung tissues,the levels of tumor necrosis factor-α( TNF-α),interleukin-1β( IL-1β) and IL-18 in BALF,the activity of NF-κB in p65 subunit,and the expression of p-JAK2 and p-STAT3 protein in the SAP group increased significantly( all P < 0. 05). Compared with the SAP group,the lung injury score,the W/D of the lung tissues,the levels of TNF-α,IL-1β,and IL-18 in BALF,the activity of NF-κB in p65 subunit,and the expression of p-JAK2 and p-STAT3 protein decreased significantly in the AG490 group,L-HSYA group and H-HSYA group( all P < 0. 05). Conclusion HSYA can attenuate the lung tissue damage and inflammatory response in SAP rats,which may be related to the inhibition of JAK2/STAT3 signaling pathway activation in the lung tissues.
引文
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