摘要
目的:探讨活性氧簇(ROS)-核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)炎症小体信号通路在丹参提取物改善急性胰腺炎肺损伤大鼠的作用机制。方法:60只SD大鼠随机分组5组:假手术组、模型组、丹参提取物(1.5 g·kg^-1)组、ROS抑制剂组[N-乙酰-L-半胱氨酸(NAC),20 mg·kg^-1]、丹参提取物+NAC组(1.5 g·kg-1+20 mg·kg^-1),每组12只。以逆行胆胰管注射5%牛磺胆酸钠(1 ml·kg^-1)法建立急性胰腺炎肺损伤大鼠模型,各组灌胃给药,模型组和假手术组给予等量生理盐水,qd,持续5 d。处死大鼠,测量腹水量、右肺干质量、右肺湿质量,计算湿质量/干质量(W/D)比值,采用苏木精-伊红(HE)染色法对大鼠肺组织病理症状进行Holfbauer评分,以全自动血气分析检测仪检测动脉血中氧分压(PaO2)、二氧化碳分压(PaCO2)、氧合指数(OI),用酶联免疫吸附(ELISA)法检测血清肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)水平,ROS检测试剂盒检测肺组织中ROS水平,蛋白免疫印迹(Western blot)法检测NLRP3、半胱氨酸蛋白酶-1(caspase-1)、凋亡相关的斑点样蛋白(ASC)表达。结果:与假手术组比较,模型组大鼠腹水量、W/D比值、Holfbauer评分、PaCO2、TNF-α、IL-6、ROS水平、NLRP3、caspase-1、ASC蛋白表达显著升高(P<0.05),PaO2、OI显著降低(P<0.05);与模型组比较,各药物处理组大鼠腹水量、W/D、Holfbauer评分、PaCO2、TNF-α、IL-6、ROS水平、NLRP3、caspase-1、ASC蛋白表达显著降低(P<0.05),PaO2、OI显著升高(P<0.05);与丹参提取物组及NAC组分别比较,丹参提取物+NAC组大鼠腹水量、W/D、Holfbauer评分、PaCO2、TNF-α、IL-6、ROS水平、NLRP3、caspase-1、ASC蛋白表达均显著降低(P<0.05),PaO2、OI显著升高(P<0.05)。结论:丹参提取物可以保护急性胰腺炎肺损伤大鼠的肺组织,可能通过下调ROS-NLRP3炎症小体信号实现。
Objective: To explore the mechanism in the improvement of reactive oxygen species( ROS)-nucleotide binding and oligomerization domain-like receptor family pyrin domain-containing 3( NLRP3) inflammatory body signaling pathway by Salvia miltiorrhiza extract in lung injury rats with acute pancreatitis. Methods: Totally 60 SD rats were randomly divided into 5 groups: the sham operation group,the model group,Salvia miltiorrhiza extract( 1. 5 g·kg^-1) group,ROS inhibitor [N-acetyl-L-cysteine( NAC),20 mg·kg^-1)]group and Salvia miltiorrhiza extract + NAC group( 1. 5 g·kg^-1+ 20 mg·kg^-1) with 12 ones in each group. The rat model of acute pancreatitis with lung injury was established by retrograde injection of 5% sodium taurocholate( 1 ml·kg^-1). Each group was given intragastric administration and the model group and the sham operation group were given the same amount of saline,once a day for 5 days. After the rats were executed,the ascites volume,dry weight of right lung and wet weight of right lung were measured,and the wet/dry weight( W/D) ratio was calculated. The Holfbauer score of pathological symptoms of lung tissue in rats was determined after hematoxylin-eosin( HE) staining. The partial pressure of oxygen( Pa O2),partial pressure of carbon dioxide( PaCO2) and oxygenation index( OI) in the arterial blood were measured by an automatic blood gas analyzer. The levels of serum tumor necrosis factor-α( TNF-α) and interleukin-6( IL-6) were detected by enzyme-linked immunosorbent assay( ELISA). The ROS assay kit was used to detect ROS level in lung tissue. Western blot was used to detect the expressions of NLRP3,caspase-1 and apoptosis-associated speck-like protein containing CARD( ASC). Results: Compared with those in the sham operation group,the ascites volume,W/D,Holfbauer score,PaCO2,TNF-α,IL-6,ROS,NLRP3,caspase-1 and ASC proteins expressions in the model group were significantly decreased,and Pa O2 and OI were significantly increased( P < 0. 05). Compared with those in the model group,the ascites volume,W/
作者
杨勇
殷红珍
胡云飞
Yang Yong;Yin Hongzhen;Hu Yunfei(Department of Emergency Internal Medicine, Chuzhou First People's Hospital, Anhui Chuzhou 239000, China;Department of ICU, Yijishan Hospital of Southern Anhui Medical College;Southern Anhui Medical College)
出处
《中国药师》
CAS
2019年第10期1795-1800,共6页
China Pharmacist
