川芎嗪对脂多糖诱导肺损伤大鼠肺组织水通道蛋白1、5表达影响的研究

发布时间：2018-07-04 15:59

  本文选题：川芎嗪 + 急性呼吸窘迫综合征　； 参考：《重庆医科大学学报》2017年01期

【摘要】：目的:初步探讨川芎嗪(tetramethylpyrazine,TMP)对脂多糖(lipopolysaccharide,LPS)诱导肺损伤大鼠肺组织水通道蛋白(aquaporins,AQPs)1、5表达的调节作用。方法:健康雄性SPF级SD大鼠32只随机分为正常对照组(C组)、TMP处理组(T组)、LPS刺激组(L组)、TMP治疗组(LT组),每组8只。L组采用尾静脉注射6 mg/kg LPS,T组给予TMP 100 mg/kg腹腔注射,LT组大鼠注射LPS 1 h后立即给予TMP 100 mg/kg腹腔注射。8 h后麻醉处死大鼠留取肺组织标本。测定肺组织湿/干重比(wet/dry ratio,W/D),苏木素-伊红(HE)染色观察肺组织病理形态,实时荧光定量PCR(quantitative real-time PCR,q RT-PCR)检测肺组织匀浆AQP1和AQP5的m RNA水平,免疫组化染色、Western blot检测肺组织AQP1、AQP5蛋白表达。结果:肺组织切片HE染色光镜下可见C组、T组肺组织结构完整、清晰;L组肺泡结构紊乱,弥漫性肺泡间隔增厚,肺泡腔内有炎性细胞浸润、出血和水肿,而LT组上述病理表现较L组减轻。L组平均肺组织湿/干重比值(5.577±0.243)明显高于其余3组(P0.01);与C、T 2组(3.430±0.251、3.251±0.142)相比,LT组湿/干重比值(4.384±0.236)增加,差异存在统计学意义(P0.01)。q RT-PCR结果显示T组AQP1、AQP5的RNA水平(1.462±0.080、1.267±0.063)高于C组(1.160±0.229、0.940±0.087),同时,L组两者转录水平下调(0.607±0.203、0.139±0.046),而LT组m RNA水平(0.848±0.071、0.314±0.134)高于L组(P0.05),免疫组化(C组:2.096±0.114、2.285±0.266;T组:3.253±0.149、3.790±0.308;L组:1.133±0.202、1.068±0.221;LT组:1.562±0.085、1.647±0.239)、Western blot结果(C组:1.443±0.103、1.278±0.061;T组:1.667±0.087、1.644±0.102;L组:0.974±0.090、0.529±0.100;LT组:1.179±0.051、0.805±0.082)与q RT-PCR一致(P0.05)。结论:川芎嗪可有效减轻脂多糖诱导急性呼吸窘迫综合征的肺水肿,起到积极的保护作用,其机制可能与上调AQP1、AQP5的表达有关。
[Abstract]:Aim: to investigate the regulatory effect of tetramethylpyrazine (TMP) on the expression of aquaporins (AQPs) in lung tissue of rats with lung injury induced by lipopolysaccharide (LPS). Methods: Thirty-two healthy male Sprague-Dawley rats with SPF grade were randomly divided into two groups: normal control group (group C) treated with TMP (group T) and LPS stimulated group (group L) (LT group). Each group (n = 8) received intraperitoneal injection of TMP100 mg/kg with tail vein injection of 6 mg/kg LPST. The rats in LT group were given TMP100 mg/kg intraperitoneally immediately after injection of LPS-1.8h later, the rats were anesthetized to collect the lung tissue. Lung tissue wet / dry weight ratio (wet/dry / W / D), hematoxylin eosin (HE) staining were used to observe the histopathology of lung tissue, the mRNA levels of AQP1 and AQP5 in lung homogenate were detected by quantitative real-time PCR, and the expression of AQP1 AQP5 protein in lung tissue was detected by immunohistochemical staining. Results: under HE staining, the lung tissue structure of group C was intact, the alveolar structure of group L was disorder, diffuse alveolar septum thickened, inflammatory cell infiltration, hemorrhage and edema were found in alveolar cavity. The mean lung wet / dry weight ratio in LT group (5.577 卤0.243) was significantly higher than that in the other three groups (P0.01), and the wet / dry weight ratio (4.384 卤0.236) in LT group was significantly higher than that in Con T2 group (3.430 卤0.251 卤3.251 卤0.142). 宸�寮傚瓨鍦ㄧ粺璁″�︽剰涔，

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