蛋白激酶抑制剂LY333531对大鼠烟雾吸入性肺损伤氧化应激的影响

Effect of protein kinase inhibitor LY333531 on oxidative stress in rats with smoke inhalation lung injury

目的探索蛋白激酶抑制剂(LY333531)对大鼠烟雾吸入后氧化应激的作用。方法 6～8周龄SD雄性SPF级大鼠40只,随机分为空白对照组、抑制剂对照组、烟雾吸入组和烟雾吸入+抑制剂组。抑制剂对照组大鼠和烟雾吸入+抑制剂组大鼠在烟雾吸入实验前1周按1 mg/(kg·d)腹腔注射LY333531。1周后,于自制产烟箱建立大鼠烟雾吸入性肺损伤模型。12 h后解剖,留取动脉血、肺组织和肺泡灌洗液。对各组大鼠进行湿干比(W/D)、肺泡灌洗液(BALF)蛋白浓度、丙二醛(MDA)、一氧化氮合酶(NOS)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和衔接蛋白(P66Shc)的检测,制作HE染色切片,在光学显微镜下观察肺组织病理学特点。结果与空白对照组和抑制剂对照组相比,烟雾吸入组大鼠肺组织W/D、肺泡灌洗液蛋白浓度均明显上升(P <0. 05);肺组织及血清中MDA、NOS浓度明显上升(P <0. 05); SOD、CAT活性明显下降(P <0. 05);衔接蛋白(P66Shc)及活化后的p-P66Shc表达明显增加;病理切片可见明显肺泡壁增厚、肺泡断裂、渗出、炎性细胞浸润等。与烟雾吸入组相比,烟雾吸入+抑制剂组大鼠肺组织W/D、肺泡灌洗液蛋白浓度均有所降低(P <0. 05);血清MDA、NOS有所降低(P <0. 05); SOD以及CAT活性增加(P <0. 05);衔接蛋白(P66Shc)及活化后的p-P66Shc表达减少;肺泡断裂、渗出、炎性细胞浸润等病理改变减轻。结论 LY333531能减轻大鼠烟雾吸入后的氧化应激效应,减轻氧化应激对机体的损伤。

Objective To explore the effect of protein kinase inhibitor(LY333531)on oxidative stress after smoke inhalation in rats.Methods Forty SD male SPF rats,6~8 weeks old,were randomly divided into the blank control group,inhibitor control group,smoke inhalation group and smoke inhalation+inhibitor group.Rats in the inhibitor control group and the smoke inhalation+inhibitor group were intraperitoneally injected with LY333531 at a dose of 1 mg/(kg·d)for one week before smoke inhalation.Then,the smoke inhalation induced acute lung injury(SI-ALI)rat model was established in a smoking generator.After 12 hours,the rats were dissected and the arterial blood,lung tissue,and bronchoalveolar lavage fluid(BALF)were collected.The wet and dry ratio(W/D),protein concentration of BALF,malondialdehyde(MDA),nitric oxide synthase(NOS),superoxide dismutase(SOD),catalase(CAT)and adaptor protein(P66 Shc)were detected.Hematoxylin-Eosin staining was performed and the histopathological features of the lung were observed under a light microscope.Results Compared with the blank control group and the inhibitor control group,the W/D of lung tissue and the protein concentration of BALF was increased significantly(P<0.05);the MDA and NOS concentrations in lung tissue and plasma increased significantly(P<0.05);the SOD and CAT activity decreased significantly(P<0.05);The expression of adaptor protein(P66 Shc)and activated p-P66 Shc was significantly increased;obvious alveolar wall thickening,alveolar rupture,exudation,and inflammatory cell infiltration were observed in pathological sections after smoke inhalation.Compared with the smoke inhalation group,the W/D and BALF protein concentrations were decreased(P<0.05).MDA and NOS in serum were decreased(P<0.05);SOD and CAT activities were increased(P<0.05);the adaptor protein(P66 Shc)and activated p-P66 Shc expression was decreased,and alveolar rupture,exudation,and inflammatory cell infiltration were alleviated with LY333531 pretreatment.Conclusion LY333531 can alleviate the oxidative stress effect after smoke inhalation in rats,and reduce the damage of oxidative stress to the body.