Effects of Chitosan on Nitric Oxide Immune Regulating Pathway in Mid-Lactating Dairy Cows

LI Tiyu; SHI Lulu; YAN Sumei; SHI Binlin

doi:10.3969/j.issn.1006-267x.2020.02.028

Chinese Journal of Animal Nutrition >

2020 , Vol. 32 >Issue 2: 726 - 735

DOI: https://doi.org/10.3969/j.issn.1006-267x.2020.02.028

RUMINANT AND HERBIVORE NUTRITION AND FEED

Effects of Chitosan on Nitric Oxide Immune Regulating Pathway in Mid-Lactating Dairy Cows

LI Tiyu ,
SHI Lulu ,
YAN Sumei ,
SHI Binlin

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College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China

Received date: 2019-08-05

Online published: 2020-02-21

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Abstract

In vivo and in vitro methods were performed in this experiment to study the effects of chitosan (CHI) on nitric oxide (NO) immune regulating pathway in mid-lactating dairy cows. Method in vivo:a total of forty heathy mid-lactating Holstein cows with similar milk yield, days in lactation, parturition date and body weight were randomly divided into five treatments, eight replicates per treatment. The five experimental diets contained, respectively, 0, 500, 1 000, 1 500 or 2 000 mg/kg CHI. Feeding trial lasted for 60 days, and was divided equally into two phases, namely 1 to 30 days (earlier period) and 31 to 60 days (later period). Cows had free access to diets and water during the trial. Method in vitro:experiment consisted of two sub-experiments and adopted 2×2×2 factor design, with 2 lipopolysaccharide (LPS) treatments (0 and 10 μg/mL), 2 chitosan oligosaccharide (COS) treatments (0 and 160 μg/mL) and 2 inhibitors (adding inhibitor and without inhibitor). There were 8 treatments in total, with 6 replicates per treatment. Two sub-experiments used respectively inducible nitric oxide synthase (iNOS) inhibitor 1400W (1 mmol/L) and nuclear transcription factor-κB (NF-κB) inhibitor PDTC (10 mmol/L). The in vivo method results showed as follows:with the increasing of the CHI addition level, NO content showed significant linear (P<0.01) or quadratic (P<0.01) increasing effects, and iNOS activity tended to be linear (0.05 ≤ P<0.10) or significant quadratic (P<0.05) increasing effects on day 60; the gene expressions of iNOS and NF-κB showed linear increasing trends (0.05 ≤ P<0.10) or significant quadratic increasing effects (P<0.05), with 1500 mg/kg being the optimum dosage in vivo. The in vitro method results showed as follows:1) when 1400W as an inhibitor, COS could significantly increase iNOS gene expression in PBMCs (P<0.05); 1400W could significantly decrease NO content and iNOS activity (P<0.01). 2) When PDTC as an inhibitor, PDTC could significantly decrease NO content (P<0.05) and gene expressions of iNOS and NF-κB (P<0.01); there were also interactions between COS and 1400W, PDTC in terms of above indicators. 3) Moreover, in LPS-stimulated conditions, LPS enhanced NO content, iNOS activity and iNOS gene expression significantly (P<0.01), and had no significant effect on NF-κB gene expression (P>0.05). Besides, adding COS could decrease NO content, iNOS activity and iNOS gene expression in comparison with in the absence of LPS stimulation. The results suggested that CHI can dose-dependently affect immune function of dairy cows by increasing gene expression level of iNOS and NF-κB, improving the activity of iNOS, stimulating the releasing of NO from immunological cells. Furthermore, COS-induced improvement of NO contents in PBMCs is a result of up-regulation of iNOS and NF-κB gene expression, and improvement of iNOS activity. COS can dual-directionally regulate key factors of NO pathway in PBMCs. In conclusion, CHI can exert immune regulation function via NO molecular pathway. The addition level of 1 500 mg/kg is considered as the optimum level of CHI.

Key words： chitosan; dairy cow; nitric oxide; immune regulation

Cite this article

LI Tiyu , SHI Lulu , YAN Sumei , SHI Binlin . Effects of Chitosan on Nitric Oxide Immune Regulating Pathway in Mid-Lactating Dairy Cows[J]. Chinese Journal of Animal Nutrition, 2020 , 32(2) : 726 -735 . DOI: 10.3969/j.issn.1006-267x.2020.02.028

References

[1] LI J,SHI B,YAN S,et al.Effects of chitosan on nitric oxide production and inducible nitric oxide synthase activity and mRNA expression in weaned piglets[J].Czech Journal of Animal Science,2015,60(8):359-366.
[2] LOWENSTEIN C J,PADALKO E.iNOS (NOS2) at a glance[J].Journal of Cell Science,2004,117(14):2865-2867.

[3] GUTIÉRREZ-VENEGAS G,VENTURA-ARROYO J A,ARREGUÍN-CANO J A,et al.Flavonoids inhibit iNOS production via mitogen activated proteins in lipoteichoic acid stimulated cardiomyoblasts[J].International Immunopharmacology,2014,21(2):320-327.

[4] GANSTER R W,TAYLOR B S,SHAO L F,et al.Complex regulation of human inducible nitric oxide synthase gene transcription by Stat 1 and NF-κB[J].Proceedings of the National Academy of Sciences of the United States of America,2001,98(15):8638-8643.

[5] 任宏,周志强,杜宪文.不同水平壳聚糖对马驹生长性能、血清激素水平和肠道菌群的影响[J].饲料研究,2019,42(4):57-59.
[6] WEI L J,LI Q,TAN W Q,et al.Synthesis,characterization,and the antioxidant activity of double quaternized chitosan derivatives[J].Molecules,2017,22(3):E501.
[7] LI J L,SHI B L,YAN S M,et al.Effects of dietary supplementation of chitosan on humoral and cellular immune function in weaned piglets[J].Animal Feed Science and Technology,2013,186(3/4):204-208.
[8] LI T,NA R,YU P,et al.Effects of dietary supplementation of chitosan on immune and antioxidative function in beef cattle[J].Czech Journal of Animal Science,2015,60(1):38-44.
[9] XU Y Q,XING Y Y,WANG Z Q,et al.Pre-protective effects of dietary chitosan supplementation against oxidative stress induced by diquat in weaned piglets[J].Cell Stress and Chaperones,2018,23(4):703-710.

[10] LI H Y,YAN S M,SHI B L,et al.Effect of chitosan on nitric oxide content and inducible nitric oxide synthase activity in serum and expression of inducible nitric oxide synthase mRNA in small intestine of broiler chickens[J].Asian-Australasian Journal of Animal Sciences,2009,22(7):1048-1053.

[11] WU G J,TSAI G J.Chitooligosaccharides in combination with interferon-γ increase nitric oxide production via nuclear factor-κB activation in murine RAW264.7 macrophages[J].Food and Chemical Toxicology,2007,45(2):250-258.

[12] WEI P,MA P,XU Q S,et al.Chitosan oligosaccharides suppress production of nitric oxide in lipopolysaccharide-induced N9 murine microglial cells in vitro[J].Glycoconjugate Journal,2012,29(5/6):285-295.
[13] PORPORATTO C,BIANCO I D,RIERA C M,et al.Chitosan induces different L-arginine metabolic pathways in resting and inflammatory macrophages[J].Biochemical and Biophysical Research Communications,2003,304(2):266-272.

[14] SCHAIRER D O,CHOUAKE J S,NOSANCHUK J D,et al.The potential of nitric oxide releasing therapies as antimicrobial agents[J].Virulence,2012,3(3):271-279.

[15] SPETH M T,REPNIK U,GRIFFITHS G.Layer-by-layer nanocoating of live Bacille-Calmette-Guérin mycobacteria with poly(I:C) and chitosan enhances pro-inflammatory activation and bactericidal capacity in murine macrophages[J].Biomaterials,2016,111:1-12.
[16] FENG J,ZHAO L H,YU Q Q.Receptor-mediated stimulatory effect of oligochitosan in macrophages[J].Biochemical and Biophysical Research Communications,2004,317(2):414-420.

[17] CAVICCHI M,WHITTLE B J R.Regulation of induction of nitric oxide synthase and the inhibitory actions of dexamethasone in the human intestinal epithelial cell line,Caco-2:influence of cell differentiation[J].British Journal of Pharmacology,1999,128(3):705-715.

[18] OZCAN L,OTUNCTEMUR A,POLAT E C,et al.Selective nuclear factor kappa B (NF-κB) inhibitor,pyrrolidium dithiocarbamate prevents,long-term histologic damage in ischemia-reperfusion injuries after delayed testicular torsion[J].Urology Journal,2016,13(3):2702-2706.
[19] JEONG H J,KOO H N,OH E Y,et al.Nitric oxide production by high molecular weight water-soluble chitosan via nuclear factor-κB activation[J].International Journal of Immunopharmacology,2000,22(11):923-933.

[20] YU Z J,ZHAO L H,KE H P.Potential role of nuclear factor-kappa B in the induction of nitric oxide and tumor necrosis factor-alpha by oligochitosan in macrophages[J].International Immunopharmacology,2004,4(2):193-200.

[21] KIM J H,KIM Y S,HWANG J W,et al.Sulfated chitosan oligosaccharides suppress LPS-induced NO production via JNK and NF-κB inactivation[J].Molecules,2014,19(11):18232-18247.

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