Regulatory effect of baicalein on intestinal ribosome biogenesis in mice with asthma

doi:10.19852/j.cnki.jtcm.2026.05.003

Abstract

Abstract:

OBJECTIVE: To investigate the therapeutic effects and mechanisms of baicalein (BAI) on asthma based on the gut-lung axis theory.

METHODS: This study utilized BALB/C mice, four-dimensional independent data acquisition proteomics, molecular docking, molecular dynamics simulation, and immunohistochemistry to explore how BAI affects ribosome biogenesis in the gut of asthmatic mice.

RESULTS: We investigated the regulatory effects of BAI on intestinal proteins, and we identified 93 differentially expressed proteins (DEPs) that overlapped among the control group, model group, and BAI (H) group. These proteins were predominantly involved in translation, cytosolic ribosomes, and structural constituents of ribosomes. BAI treatment significantly regulated the ribosome pathway. Eight ribosome biogenesis-related proteins [small ribosomal subunit protein uS9m (MRPS9), ribosomal protein L28 (RPL28), ribosomal protein L32 (RPL32), ribosomal protein S15A (RPS15A), ribosomal protein S23 (RPS23), RNA polymerase I and III subunit C (POLR1C), ribosomal protein L23A (RPL23A), and ribosomal protein S25 (RPS25)] were significantly correlated with pulmonary inflammatory factors. Molecular docking confirmed that RPS25, RPL23A, and POLR1C were primary targets of BAI. Molecular dynamics simulations revealed that the RPL23A-BAI complex exhibited greater structural stability than the RPS25-BAI and POLR1C-BAI complexes. Immunohistochemistry further verified that BAI suppressed the protein expression of RPS25, RPL23A, and POLR1C in the colon.

CONCLUSIONS: These results underscore that the mechanism by which BAI alleviates asthma may involve the regulation of ribosome biogenesis in colonic tissue.

Key words: baicalein, allergic asthma, colon, ribosome biogenesis

LU Yao, WEI Lini, WANG Ruiqi, YANG Jingwen, ZHANG Kaiqi, ZHAO Ning, LI Li. Regulatory effect of baicalein on intestinal ribosome biogenesis in mice with asthma[J]. Journal of Traditional Chinese Medicine, 2026, 46(5): 1082-1095.

Figures/Tables 10

Figure 1 Effect of BAI on airway inflammation, as determined by HE staining, mucus secretion, as determined by PAS, and inflammatory cytokine levels A: HE staining is used to detect pulmonary inflammatory infiltration (× 100); B: PAS staining is used to identify mucus secretion in lung tissue (× 100); C: HE staining is used to examine inflammatory infiltration in colon tissue (× 100); D: PAS staining is used to check the secretion of colonic mucus (× 100). A1, B1, C1, D1: Control group; A2, B2, C2, D2: Model group; A3, B3, C3, D3: BAI (L) group; A4, B4, C4, D4: BAI (M) group; A5, B5, C5, D5: BAI (H) group; A6, B6, C6, D6: Montelukast group. Control: treated only with physiological saline solution; model: treated with OVA and physiological saline; BAI (L): treated with OVA and 10 mg/kg BAI; BAI (M): treated with OVA and 20 mg/kg BAI. BAI (H): treated with OVA and 40 mg/kg BAI. BAI (L): Baicalein (low-dose); BAI (M): Baicalein (Medium-dose); BAI (H): Baicalein (high-dose). The data were analyzed with one-way analysis of variance, and the statistically significant effects were compared with Tukey’s multiple range test. HE: hematoxylin and eosin; PAS: periodic acid-schiff; OVA: ovalbumin. All the data are presented as the mean ± standard deviations ($\bar{x}$ ± s) (n = 3).

Table 1 Comparison of pathological scores of lung tissue in different groups of mice ($\bar{x}$ ± s)

Group	n	Dose (mg·kg^-1·d^-1)	HE staining score	PAS staining score
Control	3	-	0.83±0.41	2.14±0.22
Model	3	-	10.83±1.83^a	4.36±0.42^a
BAI (L)	3	10	8.83±1.94	3.21±0.13^b
BAI (M)	3	20	6.67±1.37	3.10±0.20^b
BAI (H)	3	40	3.83±0.75^b	2.76±0.32^b
Montelukast	3	10	3.83±0.75^b	2.79±0.43^b

Table 2 Special airway resistance of mice in different groups under different concentrations of Mch ($\bar{x}$± s)

Group	n	sRaw (cm H₂O.s)
Group	n	Mch (0 mg/mL)	Mch (3.125 mg/mL)	Mch (6.25 mg/mL)	Mch (12.5 mg/mL)
Control	4	2.98±0.90	3.04±0.86	3.63±0.63	4.46±0.76
Model	4	6.16±1.56^a	6.09±0.73^a	7.44±1.52^a	9.24±0.12^a
BAI (L)	4	5.24±0.55	5.45±0.29	6.13±0.17	7.49±0.84^b
BAI (M)	4	4.21±0.39^b	4.94±0.48	5.52±0.47^b	6.33±0.54^b
BAI (H)	4	3.68±0.72^b	4.68±0.75	5.25±0.68^b	5.48±0.46^b
Montelukast	4	3.56±0.68^b	4.07±1.40^b	4.56±0.93^b	5.21±1.04^b

Table 3 Effect of baicalein on the levels of IL-4, IL-5, and IL-13 in mouse lung tissue ((pg/mL, $\bar{x}$ ± s)

Group	n	IL-4	IL-5	IL-13	TSLP	IL-25	IL-33
Control	6	71.5±20.5	83.2±2.9	39.6±3.1	82.0±2.6	42.9±4.2	49.4±3.6
Model	6	629.3±66.6^a	596.7±76.4^a	141.8±14.8^a	390.4±28.2^a	232.1±15.2^a	318.3±6.0^a
BAI (L)	6	220.1±67.1^b	428.6±12.3^b	185.3±7.9	285.0±4.9^b	159.1±7.4^b	220.7±8.5^b
BAI (M)	6	174.9±46.2^b	339.5±11.4^b	101.3±8.2^b	184.9±3.8^b	132.3±5.9^b	145.2±1.7^b
BAI (H)	6	70.4±17.0^b	203.1±49.5^b	87.6±11.8^b	109.9±8.8^b	76.6±3.7^b	108.9±11.7^b
Montelukast	6	124.7±72.7^b	246.8±32.7^b	126.1±2.4^b	162.7±10.0^b	98.7±2.5^b	110.1±8.6^b

Table 4 Comparison of pathological scores of lung tissue in different groups of mice ($\bar{x}$ ± s)

Group	n	Dose (mg·kg^-1·d^-1)	HE staining score	PAS staining score
Control	3	-	0.0±0.0	7.9±1.6
Model	3	-	10.6±2.1^a	17.2±0.8^a
BAI (L)	3	10	10.6±1.1	16.2±1.9
BAI (M)	3	20	5.0±0.7	13.1±1.3
BAI (H)	3	40	1.2±0.8^b	11.2±1.2^b
Montelukast	3	10	1.4±0.6^b	10.5±0.7^b

Figure 2 Data quality monitoring and bioinformatics analysis of DEPs by category A: the ten most significantly enriched terms of the GO hierarchy and the number of involved proteins in a term; B: enriched KEGG pathways of the DEPs. The number of proteins involved and corresponding P values are shown on the right side of the column; C: PPI analysis and key node protein screening were performed with STRING and Cytoscape; D: expression heatmap of proteins involved in ribosomal biological processes; E: correlation analysis of 8 ribosome-related proteins and inflammatory factors; F: IPA to predict biological effects based on differential expression profiles in the data. The red boxes in Figures A and B, as well as the blue boxes in Figure F, refer to the content related to "ribosome biogenesis". DEPs: differentially expressed proteins; GO: gene ontology; KEGG: kyoto encyclopedia of genes and genomes; PPI: protein-protein interaction; STRING: Search Tool for the Retrieval of Interacting Genes/Proteins; IPA: ingenuity pathway analysis; BP: biological process; CC: cellular component; MF: molecular function; MRPS9: small ribosomal subunit protein uS9m; RPL28: ribosomal protein L28; RPL32: ribosomal protein L32; RPS15A: ribosomal protein S15A; RPS23: ribosomal protein S23; POLR1C: RNA polymerase I and III subunit C; RPL23A: ribosomal protein L23A; RPS25: ribosomal protein S25; IL: interleukin; TSLP: thymic stromal lymphopoietin.

Table 5 Molecular docking of 8 proteins related to the biological functions of ribosomes to baicalein

Drug	Targets	PDB ID	Binding affinity (kcal/mol)
Baicalein	RPL28	6IP8	－7.9
Baicalein	POLR1C	7OB9	－8.6
Baicalein	RPS15A	5K0Y	－6.8
Baicalein	RPS23	3MGU	－8.8
Baicalein	RPL23A	1G62	－10.5
Baicalein	RPS25	6ZOL	－9
Baicalein	RPL32	1T0K	－7.2
Baicalein	MRPS9	5AJ3	－7.5

Figure 3 MD simulation analysis of the RPS25-Baicalein, RPL23A-Baicalein, and POLR1C-Baicalein complexes for more than 100 ns. A: RMSD image; B: Rg image; C: SASA image; D: image showing the number of Hbonds; E: RMSF image (RPL23A-Baicalein); F: RMSF image (RPS25-Baicalein); G: RMSF image (POLR1C-Baicalein). E1: RPL23A-Baicalein-Chain-A; E2: RPL23A-Baicalein-Chain-B; F1: RPS25-Baicalein-Chain-A; F2: RPS25-Baicalein-Chain-B; F3: RPS25-Baicalein-Chain-C; F4: RPS25-Baicalein-Chain-D. RMSD: the root mean square deviation; Rg: the radius of gyration; SASA: solvent accessible surface area; RMSF: the root mean square fluctuation; POLR1C: RNA polymerase I and III subunit C; RPL23A: ribosomal protein L23A; RPS25: ribosomal protein S25.

Figure 4 Baicalein decreased ribosome biogenesis in the colon tissues of mice with allergic asthma A: identification of POLR1C expression in colon tissue by immunohistochemical staining (× 200), scale bar: 100 μm; B: identification of RPS25 expression in colon tissue by immunohistochemical staining (× 200), scale bar: 100 μm; C: identification of RPL23A expression in colon tissue by immunohistochemical staining (× 200), scale bar: 100 μm. A1, B1, C1: Control group; A2, B2, C2: Model group; A3, B3, C3: BAI (H) group; A4, B4, C4: Montelukast group. Control: treated only with physiological saline solution; model: treated with OVA and physiological saline; BAI (H): treated with OVA and 40 mg/kg BAI. BAI (H): Baicalein (high-dose); OVA: ovalbumin; POLR1C: RNA polymerase I and III subunit C; RPL23A: ribosomal protein L23A; RPS25: ribosomal protein S25.

Table 6 Staining intensity of POLR1C, RPS25, and RPL23A proteins in colon tissue of mice in each group (scores, $\bar{x}$ ± s)

Group	n	POLR1C/AOD	RPS25/AOD	RPL23A/AOD
Control	3	0.006±0.000	0.018±0.000	0.020±0.000
Model	3	0.024±0.000^a	0.061±0.020^a	0.073±0.020^a
BAI (H)	3	0.008±0.000^b	0.020±0.000^b	0.031±0.010^b
Montelukast	3	0.008±0.000^b	0.032±0.010	0.039±0.010

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