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目的探讨细胞外信号调节蛋白激酶1/2(ERK1/2)特异性抑制药U0126和地塞米松干预对支气管哮喘小鼠肺组织中ERK1/2磷酸化(p-ERK1/2)、白细胞介素-32(IL-32)及黏蛋白5ac(Muc5ac)蛋白表达的影响。方法将40只支气管哮喘模型小鼠随机分为模型组、实验A组、实验B组、实验C组,每组10只;另取10只正常小鼠作为对照组。模型组于第1和13天腹腔注射卵清白蛋白(OVA)混悬液0.2 mL致敏,第19天开始雾化吸入5%OVA,每次30 min,连续5 d;实验A组、实验B组及实验C组在雾化激发前30 min分别腹腔注射2mg·kg~(-1)地塞米松、30 mg·kg~(-1)U0126及二甲亚砜0.2 m L,其余操作同模型组;对照组予以0.9%NaCl,其余操作同模型组。用酶联免疫吸附试验检测肺泡灌洗液中IL-32的表达,用免疫组织化学法检测肺组织中Muc5ac、p-ERK1/2及IL-32蛋白的表达。结果模型组、实验A组、实验B组的IL-32光密度(OD)值分别为(0.81±0.25),(0.34±0.06)和(0.37±0.12),Muc5ac的积分光密度(IOD)值分别为(54462.97±6907.17),(7904.62±1639.96)和(7447.74±1233.68),p-ERK的IOD值分别为(95621.04±11112.78),(10225.53±1456.72)和(10718.69±2554.13),IL-32的IOD值分别为(44870.78±7330.11),(15075.80±3490.51)和(16847.16±3485.83)。模型组的上述指标与实验A,B组比较,差异均有统计学意义(均P<0.01)。结论 p-ERK1/2及IL-32均参与支气管哮喘小鼠气道Muc5ac的表达及分泌,且二者呈协同刺激作用。
Objective To investigate the effects of ERK1 / 2 inhibitor U0126 and dexamethasone on the expression of ERK1 / 2 phosphorylation (p-ERK1 / 2), interleukin -32 (IL-32) and mucin 5ac (Muc5ac) protein expression. Methods Forty mice with bronchial asthma were randomly divided into model group, experimental group A, experimental group B and experimental group C, with 10 mice in each group. Ten normal mice were used as control group. The model group was sensitized by intraperitoneal injection of 0.2 mL ovalbumin (OVA) suspension on day 1 and 13, and 5% OVA was inhaled on the 19th day for 30 min each for 5 consecutive days. In group A and group B Groups and experimental group C were injected intraperitoneally with dexamethasone 2 mg · kg -1, U0126 30 mg · kg -1 and dimethyl sulfoxide 0.2 ml respectively 30 min prior to atomization. Group; the control group was 0.9% NaCl, the rest of the operation with the model group. The expression of IL-32 in bronchoalveolar lavage fluid was detected by enzyme-linked immunosorbent assay, and the expression of Muc5ac, p-ERK1 / 2 and IL-32 protein in lung tissue were detected by immunohistochemistry. Results The OD values of IL-32 in model group, experimental group A and experimental group B were (0.81 ± 0.25), (0.34 ± 0.06) and (0.37 ± 0.12), respectively. The integral optical density (IOD) value of Muc5ac The IOD values of p-ERK were (95621.04 ± 11112.78), (10225.53 ± 1456.72) and (10718.69 ± 2554.13), respectively. The IOD values of p-ERK were (54462.97 ± 6907.17), (7904.62 ± 1639.96) and (7447.74 ± 1233.68) The IOD values were (44870.78 ± 7330.11), (15075.80 ± 3490.51) and (16847.16 ± 3485.83), respectively. The above indexes in model group were significantly different from those in experimental group A and group B (all P <0.01). Conclusions Both p-ERK1 / 2 and IL-32 are involved in the expression and secretion of Muc5ac in the airway of bronchial asthmatic mice, and the two are synergistic.