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目的探讨氯喹对脂多糖(LPS)所致急性肺损伤(ALI)的保护性作用及机制。方法将日龄40～50 d SD雄性大鼠96只随机分为对照组、模型组、氯喹干预组,每组各分为注射后2、4、8、12 h 4个时间点亚组。静脉注射LPS(6 mg/kg)建立ALI动物模型,氯喹干预组是在注射LPS后立即予以氯喹(30 mg/kg)腹腔注射,对照组予以静脉注射9 g/L盐水,3组注射的液体总量相等。在各观察时间点处死。测定肺湿/干质量比值(W/D),石蜡包埋切片,HE染色行肺组织病理评分,末端脱氧核苷酰基转移酶介导性dUTP切口末端标记(TUNEL)法检测肺泡巨噬细胞(AM)凋亡率,免疫组织化学法检测核因子-κBp65(NF-κBp65)表达,酶联免疫吸附法(ELISA)检测血浆IL-6水平,中性红法检测AM吞噬功能,半定量反转录-PCR(RT-PCR)法检测AM的分泌型磷脂酶A2ⅡA型(sPLA2ⅡA)、Bcl-2、Bax基因的表达。采用SPSS12.0统计软件进行分析。结果与对照组比较,模型组肺W/D、肺组织病理评分、AM的NF-κBp65表达及凋亡、AM的sPLA2ⅡA基因、Bax基因表达均显著增加(Pa<0.01),AM吞噬功能、Bcl-2基因的表达均显著下降(Pa<0.01)。与对照组比较,模型组血浆IL-6水平在注射LPS后明显升高(P<0.01)。与模型组比较,氯喹干预组上述改变得以逆转,肺损伤程度明显减轻(Pa<0.01)。结论ALI早期使用氯喹可下调AM表达sPLA2ⅡA和Bax基因,上调AM表达Bcl-2基因,减少AM凋亡,抑制NF-κB活化,增强AM吞噬功能,对LPS所致ALI有一定的保护作用。
Objective To investigate the protective effect and mechanism of chloroquine on acute lung injury (ALI) induced by lipopolysaccharide (LPS). Methods Ninety-six male Sprague-Dawley rats aged 40-50 d were randomly divided into control group, model group and chloroquine intervention group. Each group was divided into four subgroups at 2,4,8,12 h after injection. LPS (6 mg / kg) was given intravenously to establish animal model of ALI. Chloroquine group was given intraperitoneal injection of chloroquine (30 mg / kg) immediately after LPS injection, and the control group received intraperitoneal injection of 9 g / The total amount is equal. Death at each observation time point. The lung wet / dry weight ratio (W / D), paraffin-embedded sections were taken and the lung histopathological score was determined by HE staining. The expression of alveolar macrophages (TUNEL) was detected by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling AM). The expression of nuclear factor-κBp65 (NF-κBp65) was detected by immunohistochemical method. The levels of plasma IL-6 were detected by enzyme-linked immunosorbent assay (ELISA) The expression of sPLA2ⅡA, Bcl-2 and Bax in AM was detected by RT-PCR. Using SPSS12.0 statistical software for analysis. Results Compared with the control group, the lung W / D, lung histopathological score, the expression of NF-κBp65 in AM and the expression of sPLA2ⅡA and Bax in AM increased (Pa <0.01), AM phagocytosis, Bcl -2 gene expression were significantly decreased (Pa <0.01). Compared with the control group, the level of plasma IL-6 in model group was significantly increased after LPS injection (P <0.01). Compared with the model group, the above changes were reversed in the chloroquine intervention group, and the degree of lung injury was significantly reduced (Pa <0.01). Conclusions Early use of chloroquine in ALI can down-regulate the expression of sPLA2ⅡA and Bax in AM, up-regulate the expression of Bcl-2 in AM, decrease the apoptosis of AM, inhibit the activation of NF-κB, enhance the phagocytosis of AM and protect the ALI induced by LPS.