Liuwei Buqi Formula delays progression of chronic obstructive pulmonary disease in rats by regulating the NLRP3/caspase-1/GSDMD pyroptosis pathway

doi:10.12122/j.issn.1673-4254.2024.11.12

Abstract

Abstract:

Objective To explore the therapeutic mechanism of Liuwei Buqi (LWBQ) Formula for chronic obstructive pulmonary disease (COPD) in rat models. Methods SD rat models of COPD established by cigarette smoking combined with intratracheal lipopolysaccharide (LPS) instillation and hormone injection were treated with LWBQ Formula by gavage with or without intraperitoneal injection of MCC950 for 3 weeks, starting at the 5th week of modeling. After the treatments, the rats were examined for lung pathologies, lung function, total cell count and white blood cell count in bronchoalveolar lavage fluid (BALF), and serum levels of IL-6, TNF-α, IL-18 and NO. The mRNA expressions of NLRP3, ASC, caspase-1, GSDMD-N, IL-1β, and IL-18 in the lung tissue were detected with qRT-PCR. Results Compared with the normal control rats, the COPD rat models had severe lung pathologies and showed significantly decreased lung function, increased total cell and leukocyte subset counts in BALF, and increased serum levels of IL-6, TNF-α, IL-18 and NO and mRNA expressions of pyroptosis-related proteins in the lung tissue. Treatment of the rat models with LWBQ Formula significantly improved lung pathology and lung function, reduced total cell and leukocyte counts in BALF, and decreased serum levels of the inflammatory factors and expressions of pyroptosis-related proteins in the lung tissue. The combined treatment with MCC950 further improved lung pathology and function in spite of a significant difference, but BALF cell counts, serum inflammatory factor levels and pulmonary expressions of pyroptosis-related proteins were all significantly reduced following the treatment. Conclusion LWBQ Formula can delay the progression of COPD in rats possibly by inhibiting lung tissue pyroptosis via regulating the NLRP3/caspase-1/GSDMD pathway to reduce inflammatory response and lung damage.

Key words: Liuwei Buqi Formula, chronic obstructive pulmonary disease, NLRP3/caspase-1/GSDMD, pyroptosis, MCC950

Li MEI, Lu ZHANG, Di WU, Huanzhang DING, Xinru WANG, Xian ZHANG, Yuhang WEI, Zegeng LI, Jiabing TONG. Liuwei Buqi Formula delays progression of chronic obstructive pulmonary disease in rats by regulating the NLRP3/caspase-1/GSDMD pyroptosis pathway[J]. Journal of Southern Medical University, 2024, 44(11): 2156-2162.

Figures/Tables 7

Tab.1 Primer sequence

Gene	Primer sequences (5'-3')	Amplicon size(bp)
β-actin	CCCATCTATGAGGGTTACGC TTTAATGTCACGCACGATTTC	150
NLRP3	ATGCCGTATCTGGTTGTGTT GACTTAGGAAGAGCTGAGCA	142
ASC	GGCACAGCCAGAACAGAACATT TGTTTTGGTTGGGGGTCTCT	154
Caspase-1	AGCTTCAGTCAGGTCCATCAGC GGCAAAACTTGAGGGAACCAC	144
IL-1β	AGGCAGTGTCACTCATTGTGG TAGCAGGTCGTCATCATCCC	155
IL-18	TCAGACCACTTTGGCAGACT GAACACAGGCGGGTTTCTTT	96
GSDMD-N	TGAAGATCGTGGATCATGCC GGTAGAATTCCGAAGGCAGT	114

Fig.1 Comparison of lung function indexes among the groups. **P<0.01 vs control group, #P<0.05 vs model group.

Fig.2 Lung histopathological findings in each group (HE staining of rat lung tissue).

Fig.3 Giemsa staining of alveolar lavage fluid (BALF) (A) and leukocyte subset counts (B) in each group. **P<0.01 vs control group; ##P<0.01 vs model group; ΔP<0.05, ΔΔP<0.01 vs LWBQ group.

Fig.4 Comparison of serum inflammatory factors among the groups. **P<0.01 vs control group; ##P<0.01 vs model group; ΔP<0.05 vs LWBQ group.

Fig.5 Comparison of serum NO level among the groups. **P<0.01 vs control group; ##P<0.01 vs model group; ΔP<0.05 vs LWBQ group.

Fig.6 Comparison of mRNA expressions of pyroptosis-related proteins in the lung tissue among the groups. **P<0.01 vs Control group, ##P<0.01 vs model group; Δ P<0.05,ΔΔP<0.01 vs LWBQ group.

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