大蒜素改善博莱霉素诱导小鼠肺纤维化的作用分析

doi:10.3877/cma.j.issn.1674-6902.2024.01.002

目的

建立博莱霉素制备肺纤维化小鼠模型，分析大蒜素对肺纤维化的改善作用。

方法

选择C57BL/6小鼠24只，分成4组，每组6只，分别为正常对照组、正常给药组、模型组和模型给药组。模型组和模型给药组小鼠气道内单次滴注100 μl博莱霉素(浓度5 mg·kg)诱导小鼠肺纤维化。7 d后正常给药组和模型给药组每日给予100 μl大蒜素(浓度5 mg·kg)治疗，正常对照组和模型组每日给予等量生理盐水，3周后小鼠行安乐死，取肺组织备用。取部分新鲜肺组织包埋切片，HE、Masson染色，比较各组小鼠肺组织病理变化，检测肺组织羟脯氨酸(hydroxyproline, HYP)、总超氧化物歧化酶(total superoxide dismutase, T-SOD)及丙二醛(malondialdehyde, MDA)。

结果

小鼠1 d、7 d、14 d、28 d体质量：正常对照组(20.49±0.55)、(22.09±0.51)、(23.76±0.57)、(25.08±0.64)g；正常给药组(20.55±0.65)、(21.79±0.70)、(23.12±0.57)、(24.64±0.49)g；模型组(20.56±0.74)、(19.78±0.77)、(18.70±0.86)、(17.67±1.11)g；模型给药组(20.78±0.77)、(21.49±0.81)、(22.45±0.86)、(23.81±0.72)g。大蒜素治疗后小鼠肺泡炎减轻，肺纤维化程度改善。肺组织中HYP含量：正常对照组(0.87±0.09)μg/ml，正常给药组(0.86±0.03)μg/ml，模型组(1.54±0.24)μg/ml，模型给药组(0.90±0.09)μg/ml；T-SOD活性水平：正常对照组(24.20±0.11)U/mg，正常给药组(24.29±0.14)U/mg，模型组(14.75±0.64)U/mg，模型给药组(19.25±0.69)U/mg；MDA含量：正常对照组(1.21±0.08)nmol/mg，正常给药组(1.18±0.13)nmol/mg，模型组(3.23±0.07)nmol/mg，模型给药组(2.42±0.05)nmol/mg。

结论

大蒜素通过降低肺组织中HYP、MDA含量，升高T-SOD活性，对小鼠肺纤维化具有改善作用。

关键词: 肺纤维化, 大蒜素, 博莱霉素, 羟脯氨酸, 总超氧化物歧化酶, 丙二醛

Objective

To establish a bleomycin-induced pulmonary fibrosis mouse model and evaluate the therapeutic effects of allicin on pulmonary fibrosis.

Method

Twenty-four C57BL/6 mice were divided into four groups, six mice per group, as follows: normal control, normal treated, model, and model treated groups. The model and model treated mice were induced with pulmonary fibrosis by intratracheal administration of 100 μl bleomycin (5 mg·kg concentration). After seven days, both normal treated and model treated groups received daily 100 μl of allicin (5 mg·kg concentration) for treatment, while equivalent volumes of saline were administered daily to the normal control and model groups. After three weeks, the mice were euthanized, and lung tissues were harvested for analysis. Fresh lung tissue samples were embedded, sectioned, and stained with HE and Masson′s trichrome to compare the pathological changes among the different groups. Additionally, lung tissue levels of hydroxyproline (HYP), total superoxide dismutase (T-SOD) activity, and malondialdehyde (MDA) content were measured.

Result

Mouse weights at 1, 7, 14, and 28 days were: normal control group (20.49±0.55), (22.09±0.51), (23.76±0.57), (25.08±0.64)g; normal treated group (20.55±0.65), (21.79±0.70), (23.12±0.57), (24.64±0.49)g; model group (20.56±0.74), (19.78±0.77), (18.70±0.86), (17.67±1.11)g; model treated group (20.78±0.77), (21.49±0.81), (22.45±0.86), (23.81±0.72)g. Post-treatment with allicin, there was a reduction in alveolitis and significant improvement in pulmonary fibrosis. The pulmonary tissue HYP concentration values were: normal control group (0.87±0.09) μg/ml, normal treated group (0.86±0.03) μg/ml, model group (1.54±0.24) μg/ml, and model treated group (0.90±0.09) μg/ml. T-SOD activity levels: normal control group (24.20±0.11) U/mg, normal treated (24.29±0.14) U/mg, model group (14.75±0.64) U/mg, model treated group (19.25±0.69) U/mg. MDA contents: normal control group (1.21±0.08) nmol/mg, normal treated group (1.18±0.13) nmol/mg, model group (3.23±0.07) nmol/mg, model treated group (2.42±0.05) nmol/mg.

Conclusion

Allicin mitigates pulmonary fibrosis in the bleomycin-induced mouse model by reducing HYP and MDA levels and increasing T-SOD activity in lung tissue.

Key words: Pulmonary fibrosis, Allicin, Bleomycin, Hydroxyproline, Total superoxide dismutase, Malondialdehyde

表1 每组小鼠体质量变化比较[(

±s)，g]

组　别	例数	1 d	7 d	14 d	28 d
正常对照组	6	20.49±0.55	22.09±0.51^b	23.76±0.57^b	25.08±0.64^b
正常给药组	6	20.55±0.65	21.79±0.70^ab	23.12±0.57^b	24.64±0.49^b
模型组	6	20.56±0.74	19.78±0.77^a	18.70±0.86^a	17.67±1.11^a
模型给药组	6	20.78±0.77	21.49±0.81^b	22.45±0.86^b	23.81±0.72^ab

表1 每组小鼠体质量变化比较[(

±s)，g]

组　别	例数	1 d	7 d	14 d	28 d
正常对照组	6	20.49±0.55	22.09±0.51^b	23.76±0.57^b	25.08±0.64^b
正常给药组	6	20.55±0.65	21.79±0.70^ab	23.12±0.57^b	24.64±0.49^b
模型组	6	20.56±0.74	19.78±0.77^a	18.70±0.86^a	17.67±1.11^a
模型给药组	6	20.78±0.77	21.49±0.81^b	22.45±0.86^b	23.81±0.72^ab

图1 每组小鼠肺组织病理变化(HE染色，×100)。注：A：正常对照组；B：正常给药组；C：模型组；D：模型给药组

图2 每组小鼠肺组织病理变化(Masson染色，×100)。注：A：正常对照组；B：正常给药组；C：模型组；D：模型给药组

图3 大蒜素对肺纤维化小鼠肺组织HYP水平的影响

表2 大蒜素对肺纤维化小鼠肺组织T-SOD活性、MDA、HYP含量结果[(

±s)，n＝6]

组　别	例数	T-SOD(U/mg)	MDA(nmol/mg)	HYP(μg/ml)
正常对照组	6	24.20±0.11	1.21±0.08	0.87±0.09
正常给药组	6	24.29±0.14^b	1.18±0.13^b	0.86±0.03^b
模型组	6	14.75±0.64^a	3.23±0.07^a	1.54±0.24^a
模型给药组	6	19.25±0.69^ab	2.42±0.05^a	0.90±0.09^b

表2 大蒜素对肺纤维化小鼠肺组织T-SOD活性、MDA、HYP含量结果[(

±s)，n＝6]

组　别	例数	T-SOD(U/mg)	MDA(nmol/mg)	HYP(μg/ml)
正常对照组	6	24.20±0.11	1.21±0.08	0.87±0.09
正常给药组	6	24.29±0.14^b	1.18±0.13^b	0.86±0.03^b
模型组	6	14.75±0.64^a	3.23±0.07^a	1.54±0.24^a
模型给药组	6	19.25±0.69^ab	2.42±0.05^a	0.90±0.09^b

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Analysis on the effect of Allicin on bleomycin induced pulmonary fibrosis in mice

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Analysis on the effect of Allicin on bleomycin induced pulmonary fibrosis in mice