酒精戒断对脓毒症小鼠疼痛及炎症反应影响及其机制的探讨

doi:10.3877/cma.j.issn.1674-6880.2026.02.002

目的

探讨酒精戒断（AW）对脓毒症小鼠的疼痛及炎症反应的影响以及机制。

方法

30只小鼠采用平均随机法分为对照组、脓毒症组[脂多糖（LPS）组]、AW +脓毒症组（AW + LPS组），每组10只。对照组正常喂养14 d，第15天尾静脉注射等渗NaCl溶液0.1 mL；LPS组正常喂养14 d，第15天尾静脉注射LPS溶液0.1 mL；AW + LPS组采用梯度酒精喂养14 d，第15天尾静脉注射LPS溶液0.1 mL。每日观察小鼠体质量、饮水量、摄食量、行为等变化，进行疼痛评估；处死小鼠：收集肺组织，检测肺湿干重比（W/D）、诱导型一氧化氮合酶（iNOS）蛋白和信使RNA（mRNA）表达情况、观察苏木素-伊红（HE）染色病理切片；收集支气管肺泡灌洗液（BALF），检测肿瘤坏死因子α（TNF-α）及白细胞介素6（IL-6）的表达水平。

结果

与对照组相比，LPS组和AW + LPS组小鼠体质量下降、饮水和摄食量显著降低，评估为中度疼痛。HE染色显示LPS组和AW + LPS组小鼠肺部明显水肿、充血，炎症细胞渗出增加。3组小鼠肺组织W/D、iNOS的mRNA和蛋白表达以及BALF中TNF-α、IL-6水平比较，差异均有统计学意义（F = 215.400、20.270、26.900、2 318.000、985.500，P均< 0.001）；与对照组相比，LPS组和AW + LPS组小鼠上述指标均显著升高，且AW + LPS组更高（P均< 0.05）。

结论

AW加重脓毒症诱发的疼痛程度，肺损伤及全身炎症指标更加严重，且可能是通过iNOS蛋白的调节来实现的。

关键词: 脓毒症, 酒精戒断, 诱导型一氧化氮合酶蛋白, 疼痛, 炎症因子

Objective

To explore the effect and mechanism of alcohol withdrawal (AW) on the pain and inflammatory response of septic mice.

Methods

A total of 30 mice were randomly divided into a control group, a sepsis group [lipopolysaccharide (LPS) group], and an AW + sepsis group (AW + LPS group), with 10 mice in each group. Mice in the control group were fed normally for 14 days and injected with 0.1 mL isotonic NaCl solution via the tail vein on the 15th day. Mice in the LPS group received routine feeding for 14 days, followed by tail vein injection of 0.1 mL LPS solution on the 15th day. Mice in the AW + LPS group were given gradient alcohol feeding for 14 days, and injected with 0.1 mL LPS solution via the tail vein on the 15th day. Daily observations were made on the changes of mice' weight, water intake, food intake and behavior, and the pain assessments were conducted. Mice were euthanized to collect lung tissue for detecting the wet-to-dry weight ratio (W/D), measuring the protein and messenger RNA (mRNA) expression of inducible nitric oxide synthase (iNOS), and observing pathological slices via hematoxylin-eosin (HE) staining. Bronchoalveolar lavage fluid (BALF) was collected to detect the expression levels of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6).

Results

Compared with the control group, mice in the LPS group and AW + LPS group presented decreased body weight and markedly reduced water and food intake, with moderate pain. HE staining revealed obvious pulmonary edema, congestion, and increased infiltration of inflammatory cells in the lungs of mice in both the LPS group and the AW + LPS group. There were statistically significant differences in the lung W/D ratio, iNOS mRNA and protein expression, and BALF TNF-α and IL-6 levels among the three groups (F = 215.400, 20.270, 26.900, 2 318.000, 985.500; all P < 0.001). Compared with the control group, all the above indexes were significantly increased in the LPS group and AW + LPS group, with highest levels in the AW + LPS group (all P < 0.05).

Conclusion

AW aggravates sepsis-induced pain, pulmonary injury and systemic inflammatory response, which may be mediated by the regulation of iNOS protein.

Key words: Sepsis, Alcohol withdrawal, Inducible nitric oxide synthase protein, Pain, Inflammatory factor

表1 各组小鼠疼痛评估

组别	体质量	饮水、摄食量	外观	临床症状	对刺激的反应	评估
对照组	无变化	对照	无拱背、毛发竖起	无异常	敏锐	不疼痛
LPS组	较原体质量减少10%	为对照量的35%	间接性拱背、皮毛粗糙	间接性呼吸异常、震颤、痉挛	不敏锐	中度疼痛
AW + LPS组	较原体质量减少15%	为对照量的30%	间接性拱背、皮毛明显粗糙	间接性呼吸异常、震颤、痉挛	不敏锐甚至迟钝	中度疼痛

表1 各组小鼠疼痛评估

组别	体质量	饮水、摄食量	外观	临床症状	对刺激的反应	评估
对照组	无变化	对照	无拱背、毛发竖起	无异常	敏锐	不疼痛
LPS组	较原体质量减少10%	为对照量的35%	间接性拱背、皮毛粗糙	间接性呼吸异常、震颤、痉挛	不敏锐	中度疼痛
AW + LPS组	较原体质量减少15%	为对照量的30%	间接性拱背、皮毛明显粗糙	间接性呼吸异常、震颤、痉挛	不敏锐甚至迟钝	中度疼痛

表2 各组小鼠肺组织iNOS的mRNA及蛋白表达比较（

± s）

组别	只数	mRNA	蛋白
对照组	10	1.13 ± 0.19	1.00 ± 0.18
LPS组	10	1.45 ± 0.10^a	1.62 ± 0.39^a
AW + LPS组	10	1.70 ± 0.17^ab	2.15 ± 0.19^ab
F值		20.270	26.900
P值		< 0.001	< 0.001

表2 各组小鼠肺组织iNOS的mRNA及蛋白表达比较（

± s）

组别	只数	mRNA	蛋白
对照组	10	1.13 ± 0.19	1.00 ± 0.18
LPS组	10	1.45 ± 0.10^a	1.62 ± 0.39^a
AW + LPS组	10	1.70 ± 0.17^ab	2.15 ± 0.19^ab
F值		20.270	26.900
P值		< 0.001	< 0.001

图1 光镜下观察各组小鼠组织病理学变化注：LPS.脂多糖；AW.酒精戒断；HE.苏木素-伊红；a、b、c图分别为对照组、LPS组和AW + LPS组；a图显示小鼠肺组织结构清晰完整，壁薄均一、无间隔增宽，无出血、水肿、充血，无炎症细胞聚集，无液体渗出，未见明显变化；b、c图显示小鼠肺组织可见肺泡间隔明显增厚，肺泡壁变薄且断裂，肺泡腔内弥漫性充血、水肿，肺泡腔内炎症细胞大量渗出聚集；对比LPS组，AW + LPS组肺组织充血、水肿更严重（HE染色　× 40）

表3 各组小鼠BALF中TNF-α、IL-6表达水平的变化（ng/L，

± s）

组别	只数	TNF-α	IL-6
对照组	10	103.9 ± 2.1	32.9 ± 2.3
LPS组	10	301.1 ± 8.8^a	153.0 ± 8.2^a
AW + LPS组	10	505.7 ± 15.2^ab	181.2 ± 6.4^ab
F值		2 318.000	985.500
P值		< 0.001	< 0.001

表3 各组小鼠BALF中TNF-α、IL-6表达水平的变化（ng/L，

± s）

组别	只数	TNF-α	IL-6
对照组	10	103.9 ± 2.1	32.9 ± 2.3
LPS组	10	301.1 ± 8.8^a	153.0 ± 8.2^a
AW + LPS组	10	505.7 ± 15.2^ab	181.2 ± 6.4^ab
F值		2 318.000	985.500
P值		< 0.001	< 0.001

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Effect and mechanism of alcohol withdrawal on pain and inflammatory response in sepsis mice

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Effect and mechanism of alcohol withdrawal on pain and inflammatory response in sepsis mice