Protective effect and mechanism of inosine on acute lung injury induced by lipopolysaccharide in rats

doi:10.11958/20230059

Abstract

Abstract:

Objective To investigate the lung protective effect of inosine on acute lung injury (ALI) rats and its associated mechanism. Methods Twenty SD rats were randomly divided into 4 group: the control (NC) group, the model (LPS) group, the inosine low-dose intervention (IN-L) group and the inosine high-dose intervention (IN-H) group. Rats challenged with intratracheal lipopolysaccharide (LPS) were treated after 1, 6, and 12 hours with 100 mg/kg inosine (IN-L group), 200 mg/kg inosine (IN-H group) and normal saline (LPS group). The NC group received equal volume normal saline during the modeling and intervention phases. After 24 hours of the induction of ALI, the arterial blood was used to detect partial pressure of oxygen (PaO₂), the lung tissue sample was collected for the wet/dry weight (W/D) value and pathological examination. Serum and bronchoalveolar lavage fluid (BALF) were obtained to measure polyadp ADP-ribose polymerase (PARP)-1, inducible nitric oxides (iNOS), tumor necrosis factor (TNF)-α and interleukin (IL)-1β. Lung tissue homogenate was prepared to determine the toll-like receptor 4 (TLR4), myeloid differentiation primary response 88 (MYD88) and nuclear factor kappa-B (NF-κB) proteins. Results LPS induced lung tissue injury in rats, resulting in severe hypoxemia, increased levels of PARP-1 and iNOS, promoted the secretion of inflammatory factors TNF-α and IL-1β, and up-regulated expression levels of TLR4, MYD88, NF-κB proteins in lung tissue (P<0.05). Inosine intervention down-regulated expression levels of TLR4, MYD88 and NF-κB in lung tissue, decreased the production of PARP-1, iNOS, TNF-α and IL-1β, alleviated the degree of lung tissue injury and improved hypoxemia (P<0.05). The above changes were more obvious in the IN-H group than those in the IN-L group (all P<0.05). Conclusion Inosine inhibits LPS-induced lung inflammation and reduces inflammatory cytokines, which may be related in part to the inosine down-regulation of TLR4/MYD88/NF-κB signaling pathway, the inhibition of PARP-1 activity and the reduction of NF-κB nuclear translocation.

Key words: acute lung injury, lipopolysaccharide, inosine, poly (ADP-ribose) polymerase-1, NF-kappa B, toll-like receptor 4, myeloid differentiation factor 88, nitric oxide synthase

CLC Number:

R563.9，R349.6

Figures/Tables 6

References 24

[1]	ZHANG C, WANG H, WANG H, et al. A microsatellite DNA-derived oligodeoxynucleotide attenuates lipopolysaccharide-induced acute lung injury in mice by inhibiting the HMGB1-TLR4-NF-κB signaling pathway[J]. Front Microbiol, 2022, 13:964112. doi:10.3389/fmicb.2022.964112.
[2]	HE Y Q, ZHOU C C, YU L Y, et al. Natural product derived phytochemicals in managing acute lung injury by multiple mechanisms[J]. Pharmacol Res, 2021, 163:105224. doi:10.1016/j.phrs.2020.105224.
[3]	CHANG H Y, CHEN Y C, LIN J G, et al. Asatone prevents acute lung injury by reducing expressions of NF-κB,MAPK and inflammatory cytokines[J]. Am J Chin Med, 2018, 46(3):651-671. doi:10.1142/S0192415X18500349.
[4]	王莹颖, 刘文景, 宁瑶, 等. PARP抑制剂的作用机制和研究进展[J]. 中国新药杂志, 2018, 27(3):306-313.
	WANG Y Y, LIU W J, NING Y, et al. Progress in the research of PARP inhibitors and their mechanisms of action[J]. Chinese Journal of New Drugs, 2018, 27(3):306-313. doi:10.3969/j.issn.1003-3734.2018.03.011.
[5]	WASYLUK W, ZWOLAK A. PARP inhibitors:An innovative approach to the treatment of inflammation and metabolic disorders in sepsis[J]. J Inflamm Res, 2021, 14:1827-1844. doi:10.2147/JIR.S300679.
[6]	SZABO C, MARTINS V, LIAUDET L. Poly(ADP-ribose)polymerase inhibition in acute lung injury. A reemerging concept[J]. Am J Respir Cell Mol Biol, 2020, 63(5):571-590. doi:10.1165/rcmb.2020-0188TR.
[7]	弥亮钰, 吴自谦, 潘新亭, 等. PARP-1通过NF-κB信号通路对重症急性胰腺炎致肠黏膜屏障损伤的作用机制[J]. 中华急诊医学杂志, 2020, 29(5):675-681.
	MI L Y, WU Z Q, PAN X T, et al. The mechanism of poly(ADP-ribose)polymerase on the intestinal mucosal barrier injury in rat model with severe acute pancreatitis through NF-κB signaling pathway[J]. Chin J Emerg Med, 2020, 29(5):675-681. doi:10.3760/cma.j.issn.1671-0282.2020.05.013.
[8]	NASCIMENTO F P, MACEDO-JÚNIOR S J, LAPA-COSTA F R, et al. Inosine as a tool to understand and treat central nervous system disorders:A neglected actor?[J]. Front Neurosci, 2021, 15:703783. doi:10.3389/fnins.2021.703783.
[9]	LIMA G F, LOPES R O, MENDES A B A, et al. Inosine,an endogenous purine nucleoside,avoids early stages of atherosclerosis development associated to eNOS activation and p38 MAPK/NF-κB inhibition in rats[J]. Eur J Pharmacol, 2020, 882:173289. doi:10.1016/j.ejphar.2020.173289.
[10]	SRINIVASAN S, TORRES A G, POUPLANA L R. Inosine in biology and disease[J]. Genes(Basel), 2021, 12(4):600. doi:10.3390/genes12040600.
[11]	HASKÓ G, SITKOVSKY M V, SZABÓ C. Immunomodulatory and neuroprotective effects of inosine[J]. Trends Pharmacol Sci, 2004, 25(3):152-157. doi:10.1016/j.tips.2004.01.006.
[12]	LIAUDET L, MABLEY J G, PACHER P, et al. Inosine exerts a broad range of antiinflammatory effects in a murine model of acute lung injury[J]. Ann Surg, 2002, 235(4):568-578. doi:10.1097/00000658-200204000-00016.
[13]	GUO W, XIANG Q, MAO B, et al. Protective effects of microbiome-derived inosine on lipopolysaccharide-induced acute liver damage and inflammation in mice via mediating the TLR4/NF-κB pathway[J]. J Agric Food Chem, 2021, 69(27):7619-7628. doi:10.1021/acs.jafc.1c01781.
[14]	MIKAWA K, NISHINA K, TAKAO Y, et al. ONO-1714,a nitric oxide synthase inhibitor,attenuates endotoxin-induced acute lung injury in rabbits[J]. Anesth Analg, 2003, 97(6):1751-1755. doi:10.1213/01.ANE.0000086896.90343.13.
[15]	MOKRA D, MIKOLKA P, KOSUTOVA P, et al. Corticosteroids in acute lung injury:the dilemma continues[J]. Int J Mol Sci, 2019, 20(19):4765. doi:10.3390/ijms20194765.
[16]	D'ALESSIO F R. Mouse models of acute lung injury and ARDS[J]. Methods Mol Biol, 2018, 1809:341-350. doi:10.1007/978-1-4939-8570-8_22.
[17]	鲍璐璐, 崔立红. TLR4/MyD88/NF-κB信号通路的研究进展[J]. 胃肠病学和肝病学杂志, 2019, 28(5):568-572.
	BAO L L, CUI L H. Advances in TLR4/MyD88/NF-κB signaling pathway[J]. Chin J Gastroenterol Hepatol, 2019, 28(5):568-572. doi:10.3969/j.issn.1006-5709.2019.05.019.
[18]	TAKEDA K, AKIRA S. Toll-like receptors[J]. Curr Protoc Immunol, 2015, 109:14.12.1-14.12.10. doi:10.1002/0471142735.im1412s109.
[19]	WANG Y, CHE M, XIN J, et al. The role of IL-1β and TNF-α in intervertebral disc degeneration[J]. Biomed Pharmacother, 2020, 131:110660. doi:10.1016/j.biopha.2020.110660.
[20]	LIN X, KONG J, WU Q, et al. Effect of TLR4/MyD88 signaling pathway on expression of IL-1β and TNF-α in synovial fibroblasts from temporomandibular joint exposed to lipopolysaccharide[J]. Mediators Inflamm, 2015, 2015:329405. doi:10.1155/2015/329405.
[21]	郭龙, 张春媚, 高勇, 等. ARDS发病机制的相关信号通路研究进展[J]. 中国实验诊断学, 2017, 21(9):1647-1650.
	GUO L, ZHANG C M, GAO Y, et al. Progress in the signaling pathways related to the pathogenesis of ARDS[J]. Chin J Lab Diagn, 2017, 21(9):1647-1650. doi:10.3969/j.issn.1007-4287.2017.09.062.
[22]	LIU L, KE Y, JIANG X, et al. Lipopolysaccharide activates ERK-PARP-1-RelA pathway and promotes nuclear factor-κB transcription in murine macrophages[J]. Hum Immunol, 2012, 73(5):439-447. doi:10.1016/j.humimm.2012.02.002.
[23]	冯晓异, 何朋伦, 赵微, 等. 三七总皂苷对非酒精性脂肪肝大鼠的改善作用及对NO/iNOS/NF-κB通路的影响[J]. 中成药, 2021, 43(1):50-55.
	FENG X Y, HE P L, ZHAO W, et al. Effects of Panax notoginseng total saponins on improving nonalcoholic fatty liver disease rats and NO/iNOS/NF-κB signaling pathway[J]. Chinese Traditional Patent Medicine, 2021, 43(1):50-55. doi:10.3969/j.issn.1001-1528.2021.01.010.
[24]	VIRÁG L, SZABÓ C. Purines inhibit poly(ADP-ribose)polymerase activation and modulate oxidant-induced cell death[J]. FASEB J, 2001, 15(1):99-107. doi:10.1096/fj.00-0299com.

组别	病理学评分/分	动脉血PaO₂/mmHg	肺W/D
NC组	0.96±0.22	95.00±7.48	2.31±0.30
LPS组	11.92±0.46^a	45.00±6.78^a	4.94±0.18^a
IN-L组	8.20±0.49^ab	60.75±3.86^ab	4.01±0.12^ab
IN-H组	4.08±0.59^abc	73.25±3.40^abc	3.21±0.27^abc
F	563.530^＊＊	55.497^＊＊	98.249^＊＊

组别	病理学评分/分	动脉血PaO₂/mmHg	肺W/D
NC组	0.96±0.22	95.00±7.48	2.31±0.30
LPS组	11.92±0.46^a	45.00±6.78^a	4.94±0.18^a
IN-L组	8.20±0.49^ab	60.75±3.86^ab	4.01±0.12^ab
IN-H组	4.08±0.59^abc	73.25±3.40^abc	3.21±0.27^abc
F	563.530^＊＊	55.497^＊＊	98.249^＊＊

组别	血清				BALF
组别	PARP-1/ （μg/L）	iNOS/（μg/L）	TNF-α/（ng/L）	IL-1β/（ng/L）	PARP-1/ （μg/L）	iNOS/（μg/L）	TNF-α/（ng/L）	IL-1β/（ng/L）
NC组	0.28±0.07	5.51±1.12	39.39±9.16	52.38±8.27	0.18±0.06	3.27±0.65	30.93±4.99	35.36±7.44
LPS组	2.33±0.27^a	46.29±4.37^a	237.75±17.78^a	211.62±19.80^a	1.37±0.17^a	30.11±2.44^a	179.47±14.42^a	183.70±15.47^a
IN-L组	1.34±0.09^ab	26.04±2.04^ab	170.41±10.93^ab	135.60±11.42^ab	0.71±0.08^ab	16.93±1.52^ab	116.20±8.65^ab	128.53±11.28^ab
IN-H组	0.80±0.09^abc	14.18±2.33^abc	105.25±14.60^abc	86.81±11.73^abc	0.43±0.05^abc	10.95±1.43^abc	78.56±13.96^abc	55.86±6.58^abc
F	98.017^＊＊	125.035^＊＊	119.008^＊＊	78.532^＊＊	74.699^＊＊	74.699^＊＊	93.934^＊＊	119.892^＊＊

组别	血清				BALF
组别	PARP-1/ （μg/L）	iNOS/（μg/L）	TNF-α/（ng/L）	IL-1β/（ng/L）	PARP-1/ （μg/L）	iNOS/（μg/L）	TNF-α/（ng/L）	IL-1β/（ng/L）
NC组	0.28±0.07	5.51±1.12	39.39±9.16	52.38±8.27	0.18±0.06	3.27±0.65	30.93±4.99	35.36±7.44
LPS组	2.33±0.27^a	46.29±4.37^a	237.75±17.78^a	211.62±19.80^a	1.37±0.17^a	30.11±2.44^a	179.47±14.42^a	183.70±15.47^a
IN-L组	1.34±0.09^ab	26.04±2.04^ab	170.41±10.93^ab	135.60±11.42^ab	0.71±0.08^ab	16.93±1.52^ab	116.20±8.65^ab	128.53±11.28^ab
IN-H组	0.80±0.09^abc	14.18±2.33^abc	105.25±14.60^abc	86.81±11.73^abc	0.43±0.05^abc	10.95±1.43^abc	78.56±13.96^abc	55.86±6.58^abc
F	98.017^＊＊	125.035^＊＊	119.008^＊＊	78.532^＊＊	74.699^＊＊	74.699^＊＊	93.934^＊＊	119.892^＊＊

组别	TLR4	MYD88	NF-κB p65	p-NF-κB p65
NC组	0.11±0.04	0.13±0.02	0.78±0.05	0.07±0.03
LPS组	0.94±0.07^a	0.80±0.06^a	0.78±0.05	0.48±0.07^a
IN-L组	0.61±0.03^ab	0.55±0.04^ab	0.79±0.05	0.27±0.10^ab
IN-H组	0.28±0.04^abc	0.31±0.03^abc	0.78±0.04	0.15±0.09^abc
F	184.245^＊＊	149.225^＊＊	0.073	17.408^＊＊