目的:探究格隆溴铵对大鼠肺缺血再灌注损伤(LIRI)的影响,并深入分析HMGB1/TLR4/NF-κB信号通路在此过程中所发挥的作用机制,为临床应用提供理论依据。方法:选取32只6周龄的雄性SD大鼠,随机分成4个不同的组别,每组8只大鼠。4个组别分别为:假手术组(Sham)、缺血再灌注损伤模型组(I/R)、低剂量组(L)和高剂量组(H)。通过HE染色法观察并评估肺组织的病理学变化。测量湿重与干重(W/D)的比值确定肺组织的含水量。检测肺组织中与氧化应激相关的指标水平。同时,利用蛋白质印迹技术分析肺组织中HMGB1、TLR4和p-NF-κB蛋白的表达情况,采用RT-qPCR检测HMGB1、TLR4、NF-κB mRNA表达水平。结果:病理表现:Sham组肺组织结构清楚,未见损伤。I/R组肺毛细血管充血、间质水肿、肺泡壁增厚,肺组织破坏严重,有大量炎症细胞浸润。L组和H组上述情况较I/R组减轻,H组更轻。相较于Sham组,I/R组、L组的W/D比值及HMGB1、TLR4和NF-κB mRNA表达水平均有提升。然而,与I/R组相比,L组和H组在上述指标上的表现均有所下降,H组的数据表现更低,差异均具有统计学意义(P＜0.05)。进一步分析发现,相对于Sham组而言,I/R组、L组和H组的丙二醛(MDA)浓度、HMGB1、TLR4及p-NF-κB蛋白表达量均有所增加,而超氧化物歧化酶(SOD)活性则呈现下降趋势。对比I/R组,L组和H组的上述指标的表达均有所减少,同时SOD活性则有所回升,H组表现更明显,差异均具有统计学意义(P＜0.05)。结论:格隆溴铵能够通过抑制HMGB1/TLR4/NF-κB信号通路激活,减轻LIRI期间的炎症反应和氧化应激,从而改善大鼠LIRI。
Objective: To explore the effect of glycopyrronium bromide on lung ischemia-reperfusion injury (LIRI) in rats, and to analyze the mechanism of HMGB1/TLR4/NF-κB signaling pathway in this process, so as to provide theoretical basis for clinical application. Methods: Thirty-two 6-week-old male SD rats were randomly divided into 4 different groups, with 8 rats in each group. Four groups were: sham operation group (Sham), ischemia reperfusion injury model group (I/R), low dose group (L) and high dose group (H). The pathological changes of lung tissue were observed and evaluated by HE staining. The ratio of wet weight to dry weight (W/D) was measured to determine the water content of lung tissue. The levels of indicators related to oxidative stress in lung tissue were detected. At the same time, the expression of HMGB1, TLR4 and p-NF-κB protein in lung tissue was analyzed by Western blotting, and the expression levels of HMGB1, TLR4 and NF-κB mRNA were detected by RT-qPCR. Results: Pathological findings showed that the lung tissue structure of the Sham group was clear and no damage was observed; while in the I/R group, pulmonary capillary congestion, interstitial edema, alveolar wall thickening, severe lung tissue damage, and a large number of inflammatory cell infiltration were observed. The above conditions in group L and group H were less than those in the I/R group, and H group was lighter. Compared with Sham group, the W/D ratio and the expression levels of HMGB1, TLR4 and NF-κB mRNA in the I/R group and L group were increased. However, compared with the I/R group, the performance of the above indicators in the L group and H group decreased, and the data in the H group were lower, the differences were statistically significant (P＜0.05). Further analysis showed that compared with Sham group, the concentration of malondialdehyde (MDA), the expression of HMGB1, TLR4 and p-NF-κB protein in the I/R group, L group and H group increased, while the activity of superoxide dismutase (SOD) decreased. Compared with the I/R group, the expression of the above indicators in the L group and H group decreased, while the SOD activity increased, and the performance of the H group was more obvious, the differences were statistically significant (P＜0.05). Conclusion: Glycopyrronium bromide can reduce the inflammatory response and oxidative stress during LIRI by inhibiting the activation of HMGB1/TLR4/NF-κB signaling pathway, thereby improving LIRI in rats.
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