Effects of ziyuglycoside I on inflammatory injury in lung tissue of juvenile mice with mycoplasma pneumoniae via regulating the RhoA/ROCK signaling pathway

doi:10.11958/20260535

Abstract

Abstract:

Objective To investigate the effects of ziyuglycoside Ⅰ on inflammatory injury in lung tissue of juvenile mice with mycoplasma pneumoniae (MP) through regulation of the Ras homolog gene family member A (RhoA)/Rho-associated coiled-coil forming protein kinase (ROCK) pathway. Methods Seventy-two mice were randomly divided into the control group, the MP group, the low-dose ziyuglycoside Ⅰ group, the high-dose ziyuglycoside Ⅰgroup, the azithromycin group and the high-dose ziyuglycoside Ⅰ+ RhoA activator (lysophosphatidic acid) group, with 12 mice in each group. Except for the control group, all other groups were modeled by daily intranasal instillation of MP suspension for three consecutive days. After successful modeling, drug treatments were administered once daily for 7 days. Cough latency, cough frequency, partial pressure of carbon dioxide, arterial partial pressure of oxygen and lung index were detected. Pathology of lung tissue was assessed by HE staining. Levels of C-X-C motif chemokine ligand 1 (CXCL1), interleukin (IL)-1β, tumor necrosis factor-α (TNF-α) and IL-10 in lung tissue were measured by ELISA. Apoptosis in lung tissue was detected by TUNEL staining. Protein expression levels of cleaved caspase-3, P53, RhoA, ROCK1 and ROCK2 were determined by Western blot assay. Results Compared with the control group, the alveolar septa in lung tissue of the MP group were significantly widened, a large number of inflammatory cells were diffuse infiltration, the cough latency was shortened, cough frequency was increased, partial pressure of carbon dioxide was increased, and lung index, CXCL1, IL-1β, TNF-α, apoptosis rate, and protein expression of cleaved caspase-3, P53, RhoA, ROCK1 and ROCK2 were increased, while arterial partial pressure of oxygen and IL-10 levels in lung tissue were decreased (P<0.05). Compared with the MP group, the changing trends of the above indicators were opposite in the low-dose ziyuglycoside Ⅰ group, the high-dose ziyuglycoside Ⅰgroup and the azithromycin group. Moreover, the changes of each indicator were more significant in the high-dose ziyuglycoside Ⅰ group than those in the low-dose ziyuglycoside Ⅰgroup (P <0.05). There was no significant difference between the high-dose ziyuglycoside Ⅰ group and the azithromycin group (P > 0.05). Lysophosphatidic acid reversed the ameliorative effect of high-dose ziyuglycoside Ⅰon pulmonary inflammatory injury in MP-infected juvenile mice. Conclusion Ziyuglycoside I can effectively ameliorate inflammatory lung injury in MP of juvenile mice, and its mechanism may be related to the inhibition of RhoA/ROCK pathway activation.

Key words: pneumonia, mycoplasma, inflammation, apoptosis, rho-associated kinases, Ziyuglycoside Ⅰ, RhoA/ROCK pathway

CLC Number:

R563.1

Figures/Tables 7

References 24

[1]	He J, Tian W, Feng P, et al. Nasal delivery of Lacticaseibacillus rhamnosus GG modulates respiratory immune responses and attenuates Mycoplasma pneumoniae pneumonia in a murine model[J]. Microbiol Spectr, 2026, 14(1):e00771-e00725. doi:10.1128/spectrum.00771-25.
[2]	Zhou Y, Su X, Wang C, et al. Toosendanin attenuates Mycoplasma pneumonia induced pneumonia (MPP) in mice via inhibiting NF-κb-mediated inflammatory response:in vivo and silico studies[J]. Dose Response, 2025, 23(3):15593258251367619. doi:10.1177/15593258251367619.
[3]	Kim K, Jung S, Kim M, et al. Global trends in the proportion of macrolide-resistant Mycoplasma pneumoniae infections:a systematic review and meta-analysis[J]. JAMA Netw Open, 2022, 5(7):e2220949. doi:10.1001/jamanetworkopen.2022.20949.
[4]	Song Z, Han C, Luo G, et al. Yinqin Qingfei granules alleviate Mycoplasma pneumoniae pneumonia via inhibiting NLRP3 inflammasome-mediated macrophage pyroptosis[J]. Front Pharmacol, 2024, 15:1437475. doi:10.3389/fphar.2024.1437475.
[5]	Hossen F, Geng X, Sun G Y, et al. Oligomeric amyloid-β and tau alter cell adhesion properties and induce inflammatory responses in cerebral endothelial cells through the RhoA/ROCK pathway[J]. Mol Neurobiol, 2024, 61(11):8759-8776. doi:10.1007/s12035-024-04138-z.
[6]	严娅芬, 陆光兵, 田良东, 等. 橄榄苦苷调节RhoA/ROCK信号通路对肺炎链球菌感染导致的肺泡上皮细胞损伤的影响[J]. 中国细胞生物学学报, 2024, 46(9):1663-1671.
	Yan Y F, Lu G B, Tian L D, et al. Effect of oleuropein on alveolar epithelial cell injury induced by streptococcus pneumoniae infection by regulating the RhoA/ROCK signaling pathway[J]. Chinese Journal of Cell Biology, 2024, 46(9):1663-1671. doi:10.11844/cjcb.2024.09.0006.
[7]	包代琴, 刘奕言, 张紫森, 等. 地榆皂苷Ⅰ对脓毒症大鼠急性肺损伤的保护作用及机制研究[J]. 中国药房, 2023, 34(5):537-543.
	Bao D Q, Liu Y Y, Zhang Z S, et al. Study on protective effects and mechanism of ziyuglycoside Ⅰ on acute lung injury in sepsis rats[J]. China Pharmacy, 2023, 34(5):537-543. doi:10.6039/j.issn.1001-0408.2023.05.05.
[8]	林杉, 王云, 刘哲, 等. 圣草酚调节IL-6/STAT3信号通路对支原体肺炎幼鼠肺组织的保护作用[J]. 中国细胞生物学学报, 2025, 47(8):1992-2000.
	Lin S, Wang Y, Liu Z, et al. The protective effect of eriodictyol on lung tissue of young mice with mycoplasma pneumonia by regulating IL-6/STAT3 signaling pathway[J]. Chinese Journal of Cell Biology, 2025, 47(8):1992-2000. doi:10.11844/cjcb.2025.08.0022.
[9]	马鸿菲, 唐静, 尹俪桥, 等. 发酵桔梗调控EGFR/MAPK/ERK信号通路修复支原体肺炎肺损伤[J]. 中药药理与临床, 2026, 42(2):90-97.
	Ma H F, Tang J, Yin L Q, et al. Fermented jiegeng regulates EGFR/MAPK/ERK signaling pathway to repair lung injury causedby mycoplasmal pneumonia[J]. Pharmacology and Clinics of Chinese Materia Medica, 2026, 42(2):90-97. doi:10.13412/j.cnki.zyyl.20250522.002.
[10]	张涛, 毛治尉, 豆倩云, 等. 基于RhoA/ROCK通路探讨赤芍总苷对心肌梗死大鼠心肌凋亡的影响[J]. 中国老年学杂志, 2023, 43(5):1138-1142.
	Zhang T, Mao Z W, Dou Q Y, et al. Effect of total glucosides of red peony on myocardial apoptosis in rats with myocardial infarction based on the RhoA/ROCK pathway[J]. Chinese Journal of Gerontology, 2023, 43(5):1138-1142. doi:10.3969/j.issn.1005-9202.2023.05.033.
[11]	Yuan Y, Zhou Y. NNT-AS1,a long non-coding RNA with therapeutic promise in mycoplasma pneumoniae pneumonia via the miR-410-3p/TMEM14A/Wnt/β-catenin signalling pathway[J]. Iran J Biotechnol, 2025, 23(1):e4034. doi:10.30498/ijb.2025.492048.4034.
[12]	Long H, He G, He J, et al. The protective effect and immunomodulatory ability of orally administrated Lacticaseibacillus rhamnosus GG against Mycoplasma pneumoniae infection in BALB/c mice[J]. PLoS One, 2024, 19(10):e0312318. doi:10.1371/journal.pone.0312318.
[13]	Sang M, Wang J, Jiang X, et al. A study on the influencing factors of chronic cough caused by Mycoplasma pneumoniae[J]. Pediatr Pulmonol, 2025, 60(10):e71284. doi:10.1002/ppul.71284.
[14]	Sun X, Xiang H, Liu Z, et al. Jingfang Granules alleviates bleomycin-induced acute lung injury through regulating PI3K/Akt/mTOR signaling pathway[J]. J Ethnopharmacol, 2024, 318(Pt A):116946. doi:10.1016/j.jep.2023.116946.
[15]	Jin S, Huang B, Kong Y, et al. Effect of neuromuscular electrical stimulation combined with respiratory rehabilitation training on pulmonary rehabilitation in patients with chronic obstructive pulmonary disease[J]. J Cardiothorac Surg, 2025, 20(1):79. doi:10.1186/s13019-024-03329-y.
[16]	Zhu M, Lu Y, Wei Y, et al. The NLRP3 inflammasome activation boosts lung injury,inflammation,and macrolide resistance in Mycoplasma pneumoniae pneumonia[J]. Cytokine, 2025, 195:157014. doi:10.1016/j.cyto.2025.157014.
[17]	Song D, Wei W, Zhang J, et al. The mechanism of baicalin in improving pulmonary inflammatory response and injury and regulating intestinal flora in Mycoplasma pneumoniae pneumonia mice[J]. Cell Signal, 2025, 126:111530. doi:10.1016/j.cellsig.2024.111530.
[18]	Chen C, Hu J, Guo M, et al. LncNFYC-AS1 ameliorates Mycoplasma pneumoniae pneumonia via regulating miR-1323[J]. BMC Pulm Med, 2025, 25(1):457. doi:10.1186/s12890-025-03926-0.
[19]	Zeng Q, Sun P, Li W, et al. Protective immunity induced by a novel P1 adhesin C-terminal anchored mRNA vaccine against Mycoplasma pneumoniae infection in BALB/c mice[J]. Microbiol Spectr, 2025, 13(3):e02140-e02124. doi:10.1128/spectrum.02140-24.
[20]	Wang L, Liu L, Cheng S, et al. In vitro and in vivo study of andrographolide nanoparticles for the treatment of Mycoplasma pneumoniae pneumonia[J]. Biochem Biophys Res Commun, 2024, 698:149540. doi:10.1016/j.bbrc.2024.149540.
[21]	Yan H, Li H, Yin D H, et al. The PIWI-interacting RNA CRAPIR alleviates myocardial ischemia-reperfusion injury by reducing p53-mediated apoptosis via binding to SRSF1[J]. Acta Pharmacol Sin, 2025, 46(9):2393-2406. doi:10.1038/s41401-025-01534-6.
[22]	Sun H, Wang M, Su T, et al. Ziyuglycoside I attenuates collagen-induced arthritis through inhibiting plasma cell expansion[J]. J Ethnopharmacol, 2022, 294:115348. doi:10.1016/j.jep.2022.115348.
[23]	Wang H, Chen L, Li R, et al. Polydopamine-coated mesoporous silica nanoparticles co-loaded with Ziyuglycoside Ⅰand Oseltamivir for synergistic treatment of viral pneumonia[J]. Int J Pharm, 2023, 645:123412. doi:10.1016/j.ijpharm.2023.123412.
[24]	Lv Y, Chen Y, Gao Z, et al. Diesel exhaust particles induced blood-brain barrier dysfunction through inflammation,oxidative stress,and activation of the RhoA/ROCK signaling pathway[J]. Neurotoxicology, 2025, 110:122-131. doi:10.1016/j.neuro.2025.08.002.

组别	咳嗽潜伏期/s	咳嗽次数/次
对照组	88.67±7.04	14.23±0.95
MP组	34.46±2.09^a	53.68±3.18^a
地榆皂苷Ⅰ小剂量组	45.73±4.06^b	44.76±4.09^b
地榆皂苷Ⅰ大剂量组	73.65±5.21^bc	25.66±2.28^bc
阿奇霉素组	74.22±6.08^bc	27.15±3.07^bc
地榆皂苷Ⅰ大剂量+溶血磷脂酸组	61.22±4.05^d	35.26±4.03^d
F	192.200^＊＊	248.200^＊＊

组别	咳嗽潜伏期/s	咳嗽次数/次
对照组	88.67±7.04	14.23±0.95
MP组	34.46±2.09^a	53.68±3.18^a
地榆皂苷Ⅰ小剂量组	45.73±4.06^b	44.76±4.09^b
地榆皂苷Ⅰ大剂量组	73.65±5.21^bc	25.66±2.28^bc
阿奇霉素组	74.22±6.08^bc	27.15±3.07^bc
地榆皂苷Ⅰ大剂量+溶血磷脂酸组	61.22±4.05^d	35.26±4.03^d
F	192.200^＊＊	248.200^＊＊

组别	二氧化碳分压/ mmHg	动脉血氧分压/ mmHg	肺指数/ %
对照组	40.67±3.01	105.23±6.93	0.43±0.05
MP组	98.85±6.12^a	51.67±3.28^a	1.05±0.12^a
地榆皂苷Ⅰ小剂量组	81.73±5.03^b	63.78±5.02^b	0.89±0.09^b
地榆皂苷Ⅰ大剂量组	54.67±3.41^bc	97.64±6.26^bc	0.61±0.07^bc
阿奇霉素组	55.21±3.52^bc	98.22±6.08^bc	0.63±0.08^bc
地榆皂苷Ⅰ大剂量+ 溶血磷脂酸组	68.27±4.51^d	81.61±5.02^d	0.73±0.09^d
F	274.700^＊＊	178.200^＊＊	78.530^＊＊

组别	二氧化碳分压/ mmHg	动脉血氧分压/ mmHg	肺指数/ %
对照组	40.67±3.01	105.23±6.93	0.43±0.05
MP组	98.85±6.12^a	51.67±3.28^a	1.05±0.12^a
地榆皂苷Ⅰ小剂量组	81.73±5.03^b	63.78±5.02^b	0.89±0.09^b
地榆皂苷Ⅰ大剂量组	54.67±3.41^bc	97.64±6.26^bc	0.61±0.07^bc
阿奇霉素组	55.21±3.52^bc	98.22±6.08^bc	0.63±0.08^bc
地榆皂苷Ⅰ大剂量+ 溶血磷脂酸组	68.27±4.51^d	81.61±5.02^d	0.73±0.09^d
F	274.700^＊＊	178.200^＊＊	78.530^＊＊

组别	CXCL1	IL-1β
对照组	20.68±1.57	17.23±1.09
MP组	196.88±14.13^a	145.67±9.02^a
地榆皂苷Ⅰ小剂量组	161.72±10.06^b	118.73±7.15^b
地榆皂苷Ⅰ大剂量组	76.62±5.47^bc	42.69±3.27^bc
阿奇霉素组	71.21±5.09^bc	40.28±3.11^bc
地榆皂苷Ⅰ大剂量+溶血磷脂酸组	131.26±10.55^d	79.67±5.08^d
F	325.500^＊＊	500.800^＊＊
组别	TNF-α	IL-10
对照组	40.47±3.05	97.45±5.17
MP组	234.06±15.12^a	21.22±1.45^a
地榆皂苷Ⅰ小剂量组	200.84±11.07^b	36.19±2.71^b
地榆皂苷Ⅰ大剂量组	98.65±5.04^bc	80.64±5.11^bc
阿奇霉素组	95.77±5.08^bc	81.23±5.09^bc
地榆皂苷Ⅰ大剂量+溶血磷脂酸组	143.74±12.04^d	55.75±3.01^d
F	336.500^＊＊	321.500^＊＊