肉桂精油及肉桂醛的体外抗炎活性及对LPS诱导RAW 264.7细胞炎症损伤的保护作用

doi:10.11843/j.issn.0366-6964.2025.10.026

摘要/Abstract

摘要：

旨在探究肉桂精油(CEO)与肉桂醛(CIN)的体外抗炎活性以及对细胞炎症损伤的保护作用，为CEO和CIN发挥其抗炎活性作用应用于畜禽生产提供理论参考。本研究包括2个试验：1)体外化学抗炎活性试验。通过ELISA法测定CEO和CIN(分别设置4种不同浓度：6.25、12.5、25、50 mg·mL^-1)对炎症相关酶(环氧合酶-2(COX-2)与5-脂氧合酶(5-LOX))的抑制作用，评价2种植物精油的抗炎活性；2)脂多糖(LPS)诱导RAW264.7细胞炎症损伤试验。先进行CEO和CIN对RAW264.7细胞毒性试验，以确定本试验的CEO和CIN试验浓度。再设置对照组、LPS组和植物精油组，其中，LPS组细胞正常培养后加入1 μg·mL^-1 LPS处理24 h，CEO和CIN组细胞正常培养后先加入不同浓度(1、3、5 μg·mL^-1)的CEO或CIN处理12 h，再加入1 μg·mL^-1 LPS处理24 h。体外抗炎试验结果表明：CEO和CIN对COX-2和5-LOX具有抑制活性，且CEO和CIN在浓度6.25~50 mg·mL^-1均能呈剂量依赖性显著降低COX-2(P_L < 0.01)和5-LOX(P_Q < 0.01)的含量，在相同浓度下，CEO对COX-2和5-LOX的抑制作用显著高于CIN(P < 0.01)。细胞试验结果表明：1)细胞毒性试验发现，CEO和CIN在浓度为0.781 25~6.25 μg·mL^-1时显著提高细胞活力(P < 0.05)，因此选择1、3、5 μg·mL^-1进行后续试验；2)对细胞形态观察的结果表明，与LPS组相比，1、3、5 μg·mL^-1的CEO和CIN均能不同程度地降低RAW 264.7细胞形态的分化；3)与LPS组相比，3、5 μg·mL^-1的CEO和CIN预处理后均呈二次曲线显著降低RAW264.7细胞中一氧化氮(NO)含量(P_Q < 0.05)，且CEO的抑制作用显著高于CIN(P < 0.01)；4)与LPS组相比，1、3、5 μg·mL^-1的CEO和CIN预处理均显著降低RAW264.7细胞内白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)的含量(P < 0.05)。此外，3、5 μg·mL^-1的CEO降低IL-1β和TNF-α含量的能力显著高于CIN(P < 0.01)，但对IL-6含量的影响无显著差异(P＞0.05)。综上所述，CEO和CIN均具有良好的抗炎活性，在本试验条件下，CEO对COX-2与5-LOX的抑制作用及对LPS诱导RAW264.7细胞炎症的保护作用均强于CIN，表明CEO具有更强的抗炎活性。

关键词: 肉桂精油, 肉桂醛, COX-2, 5-LOX, RAW264.7细胞

Abstract:

The purpose of this study was to explore the anti-inflammatory activity of cinnamon essential oil (CEO) and cinnamaldehyde (CIN) and the protective effect on cellular inflammatory damage, and to provide theoretical reference for CEO and CIN to exert anti-inflammatory activity in livestock and poultry production. This study included 2 trials: 1) In vitro chemical anti-inflammatory study, the anti-inflammatory activity of CEO and CIN at four concentrations (6.25, 12.5, 25, 50 mg ·mL^-1) by measuring the inhibition of inflammation-related enzymes (cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX)) by ELISA method. 2) In the study of LPS induced inflammatory damage in RAW264.7 cells, the cytotoxicity tests were performed first to determine the CEO and CIN concentrations. Next, the RAW264.7 cells in the control group were cultured in a normal medium without any treatment. The cells in the LPS group were cultured in a normal medium and treated with LPS for 24 h. The cells in the CEO and CIN groups were cultured in a normal medium and treated with CEO or CIN of different concentrations (1, 3 and 5 μg·mL^-1) for 12 h, and subsequently treated with LPS for 24 h. The in vitroresults showed that both CEO and CIN exhibited dual inhibitory activity against COX-2/5-LOX, and showed dose-dependent reduction of COX-2 (P_L < 0.01) and 5-LOX (P_Q < 0.01) at 6.25-50 mg·mL^-1, and the inhibitory effect of CEO was significantly higher than (P < 0.01) CIN at the same concentration. The cell test results were showed as follows: 1) The results of cytotoxicity tests showed that CEO and CIN significantly increased (P < 0.05) cell survival rate within 0.781 25-6.25 μg·mL^-1, and 1, 3, and 5 μg·mL^-1 were selected for subsequent tests. 2) The observation of RAW 264.7 cell morphology showed that 1, 3 and 5 μg·mL^-1 CEO and CIN pretreatment could reduce the differentiation to varying degrees compared with the LPS group. 3) Compared with the LPS group, nitric oxide (NO) content was significantly decreased (P_Q < 0.05) in 3 and 5 μg·mL^-1 CEO and CIN groups, and the inhibitory effect of CEO was significantly higher than CIN (P < 0.01). 4) Compared with LPS group, the content of interleukin-1 β (IL-1 β), interleukin-6 (IL-6), and TNF-α (TNF-α) were significantly reduced (P < 0.05) in 1, 3 and 5 μg·mL^-1 CEO and CIN groups. In addition, the 3, 5 μg·mL^-1 CEO had significantly higher (P < 0.01) ability to reduce IL-1β and TNF-α content than CIN, but there was no significant difference on IL-6 (P>0.05). In conclusion, CEO and CIN had excellent anti-inflammatory activity, and the inhibitory effect of CEO on COX-2 and 5-LOX and the protective effect of LPS-induced inflammation in RAW264.7 cells were significantly stronger than that of CIN under the conditions of the current study.

Key words: cinnamon essential oil, cinnamaldehyde, COX-2, 5-LOX, RAW264.7 cells

中图分类号:

S816.7

张岩, 欧念涛, 刘孟哲, 王凯, 贾慧鑫, 于文静, 李艳玲. 肉桂精油及肉桂醛的体外抗炎活性及对LPS诱导RAW 264.7细胞炎症损伤的保护作用[J]. 畜牧兽医学报, 2025, 56(10): 5060-5071.

ZHANG Yan, OU Niantao, LIU Mengzhe, WANG Kai, JIA Huixin, YU Wenjing, LI Yanling. In vitro Anti-inflammatory Activity of Cinnamon Essential Oil and Cinnamaldehyde, and the Protective Effect against LPS-induced Inflammatory Injury in RAW 264.7 cells[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(10): 5060-5071.

图/表 6

表 1

表 2

图 1

图 2

图 3

图 4

参考文献 49

1	NIUX,DINGY,CHENS,et al.Effect of immune stress on growth performance and immune functions of livestock: mechanisms and prevention[J].Animals (Basel),2022,12(7):909.
2	MEDZHITOVR.Inflammation 2010: new adventures of an old flame[J].Cell,2010,140(6):771-776. doi: 10.1016/j.cell.2010.03.006
3	PJ J,MANJUS L,ETHIRAJK R,et al.Safer anti-inflammatory therapy through dual COX-2/5-LOX inhibitors: A structure-based approach[J].Eur J Pharm Sci,2018,121,356-381. doi: 10.1016/j.ejps.2018.06.003
4	HORTELANOS,ZEINIM,BOSCAL,et al.Nitric oxide and cell viability in inflammatory cells: A role for NO in macrophage function and fate[J].Toxicol,2005,208(2):249-258. doi: 10.1016/j.tox.2004.11.035
5	SINGHB,SINGHJ P,KAURA,et al.Insights into the chemical composition and bioactivities of citrus peel essential oils[J].Food Res. Int,2021,143,110231. doi: 10.1016/j.foodres.2021.110231
6	PLASTINAP,APRIANTINIA,MEIJERINKJ,et al.In vitroanti-inflammatory and radical scavenging properties of Chinotto (Citrus myrtifolia Raf.) essential oils[J].Nutrients,2018,10(6):783. doi: 10.3390/nu10060783
7	SHENO Y.Anti-inflammatory effect of essential oil from Citrus aurantium L. var.amara Engl[J].J Agric Food Chem,2017,65(39):8586-8594. doi: 10.1021/acs.jafc.7b02586
8	STEVANOVÍCZ,BOŠNJAK-NEUMVLLERJ,PAJÍC-LIJAKOVÍCI,et al.Essential oils as feed additives—future perspectives[J].Molecules,2018,23(7):1717. doi: 10.3390/molecules23071717
9	NEHMER,ANDRESS,PEREIRAR B,et al.Essential oils in livestock: From health to food quality[J].Antioxidants (Basel),2021,10(2):330. doi: 10.3390/antiox10020330
10	张嘉琦,张会艳,赵青余,等.植物精油对畜禽肠道健康、免疫调节和肉品质的研究进展[J].动物营养学报,2021,33(5):2439-2451.
	ZHANGJ Q,ZHANGH Y,ZHAOQ Y,et al.Research progress of plant essential oil on intestinal health, immune regulation and meat quality of livestock and poultry[J].Journal of Animal Nutrition,2021,33(5):2439-2451.
11	SHUC,GEL,LIZ,et al.Antibacterial activity of cinnamon essential oil and its main component of cinnamaldehyde and the underlying mechanism[J].Front Pharmacol,2024,15,1378434. doi: 10.3389/fphar.2024.1378434
12	LIUS,ZHAOC,CAOY,et al.Comparison of chemical compositions and antioxidant activity of essential oils from litsea cubeba, cinnamon, Anise, and Eucalyptus[J].Molecules,2023,28(13):5051. doi: 10.3390/molecules28135051
13	ZHAOC,CAOY,ZHANGZ,et al.Cinnamon and eucalyptus oils suppress the inflammation induced by lipopolysaccharide in vivo[J].Molecules,2021,26(23):7410. doi: 10.3390/molecules26237410
14	CHOWDHURYS,MANDALG P,PATRAA K,et al.Different essential oils in diets of broiler chickens: 2. Gut microbes and morphology, immune response, and some blood profile and antioxidant enzymes[J].Anim Feed Sci Technol,2017,236,39-47.
15	PALHARESC J,GESTEIRAC S,BELLIA L,et al.Effects of a blend of essential oils in milk replacer on performance, rumen fermentation, blood parameters, and health scores of dairy heifers[J].PLoS One,2021,16(3):e231068.
16	OOIL S,LIY,KAMS L,et al.Antimicrobial activities of cinnamon oil and cinnamaldehyde from the Chinese medicinal herb cinnamomum cassia blume[J].Am J Chin Med,2006(3):34, 511-522.
17	CHANGK S,TAKJ H,KIMS I,et al.Repellency of Cinnamomum cassiabark compounds and cream containing cassia oil to Aedes aegypti(Diptera: Culicidae) under laboratory and indoor conditions[J].Pest Manag Sci,2006,62(11):1032-1038. doi: 10.1002/ps.1268
18	LUL,SHUC,CHENL,et al.Insecticidal activity and mechanism of cinnamaldehyde inC. elegans[J].Fitoterapia,2020,146,104687. doi: 10.1016/j.fitote.2020.104687
19	JIMENEZM J,BERRIOSR,STELZHAMMERS,et al.Ingestion of organic acids and cinnamaldehyde improves tissue homeostasis of piglets exposed to enterotoxic Escherichia coli(ETEC)[J].J Anim Sci,2020,98(2):skaa012. doi: 10.1093/jas/skaa012
20	SAMPATHC,CHUKKAPALLIS S,RAJUA V,et al.Cinnamaldehyde protects againstP. gingivalis induced intestinal epithelial barrier dysfunction in IEC-6 cells via the PI3K/Akt-Mediated NO/Nrf2 signaling pathway[J].Int J Mol Sci,2024,25(9):4734. doi: 10.3390/ijms25094734
21	NGUYENL,NGUYENN,TRANK N,et al.Anxiolytic-like effect of inhaled cinnamon essential oil and its main component cinnamaldehyde in animal models[J].Molecules,2022,27(22):7997. doi: 10.3390/molecules27227997
22	SIMJ,KHAZANDIM,PIH,et al.Antimicrobial effects of cinnamon essential oil and cinnamaldehyde combined with EDTA against canine otitis externa pathogens[J].J Appl Microbiol,2019,127(1):99-108. doi: 10.1111/jam.14298
23	STEVENSN,ALLREDK.Antidiabetic potential of volatile cinnamon oil: A review and exploration of mechanisms using in silico molecular docking simulations[J].Molecules,2022,27(3):853. doi: 10.3390/molecules27030853
24	VENDRAMINI-COSTAD B,CARVALHOJ E.Molecular link mechanisms between inflammation and cancer[J].Curr Pharm Des,2012,18(26):3831-3852. doi: 10.2174/138161212802083707
25	YARLAN S,BISHAYEEA,SETHIG,et al.Targeting arachidonic acid pathway by natural products for cancer prevention and therapy[J].Semin Cancer Biol,2016,40-41,48-81. doi: 10.1016/j.semcancer.2016.02.001
26	SIMONL S.Role and regulation of cyclooxygenase-2 during inflammation[J].Am J Med,1999,106(5):37S-42S. doi: 10.1016/S0002-9343(99)00115-1
27	FURIAE,NAPOLIA,TAGARELLIA,et al.Speciation of 2-Hydroxybenzoic acid with calcium(Ⅱ), magnesium(Ⅱ), and nickel(Ⅱ) cations in self-medium[J].J Chem Eng Data,2013,58(5):1349-1353. doi: 10.1021/je400105c
28	CERELLAC,SOBOLEWSKIC,DICATOM,et al.Targeting COX-2 expression by natural compounds: a promising alternative strategy to synthetic COX-2 inhibitors for cancer chemoprevention and therapy[J].Biochem Pharmacol,2010,80(12):1801-1815. doi: 10.1016/j.bcp.2010.06.050
29	MASHIMAR,OKUYAMAT.The role of lipoxygenases in pathophysiology; new insights and future perspectives[J].Redox Biol,2015,6,297-310. doi: 10.1016/j.redox.2015.08.006
30	ZHENGM,ZHANGZ,ZHUW,et al.Essential structural profile of a dual functional inhibitor against cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX): molecular docking and 3D-QSAR analyses on DHDMBF analogues[J].Bioorg Med Chem,2006,14(10):3428-3437. doi: 10.1016/j.bmc.2005.12.062
31	BILTEKINS N,DEMIRCIF.In vitroanti-inflammatory and anticancer evaluation of Mentha spicata L. essential oils[J].Adv Mater,2023,8(19):17143-17150.
32	KHANA,PERVAIZA,ANSARIB,et al.Phytochemical profiling, anti-inflammatory, anti-oxidant and in-silico approach of Cornus macrophyllabioss (bark)[J].Molecules,2022,27(13):4081. doi: 10.3390/molecules27134081
33	王雪,刘燕,史玉柱,等.瘤果黑种草子总皂苷的抗炎和免疫调节作用[J].免疫学杂志,2023,39(2):120-127.
	WANGX,LIUY,SHIY Z,et al.The anti-inflammatory and immunomodulatory effects of total saponins from seeds of nigella glandulifera[J].Immunological Journal,2023,39(2):120-127.
34	BOGDANC.Nitric oxide and the immune response[J].Nat Immunol,2001,2(10):907-916. doi: 10.1038/ni1001-907
35	MONCADAS,HIGGSA.The L-arginine-nitric oxide pathway[J].N Engl J Med,1993,329(27):2002-2012. doi: 10.1056/NEJM199312303292706
36	TUNGY T,CHUAM T,WANGS Y,et al.Anti-inflammation activities of essential oil and its constituents from indigenous cinnamon (Cinnamomum osmophloeum) twigs[J].Bioresour Technol,2008,99(9):3908-3913. doi: 10.1016/j.biortech.2007.07.050
37	LIUC,CHUD,KALANTAR-ZADEHK,et al.Cytokines: from clinical significance to quantification[J].Adv Sci (Weinh),2021,8(15):e2004433. doi: 10.1002/advs.202004433
38	LEES C,WANGS Y,LIC C,et al.Anti-inflammatory effect of cinnamaldehyde and linalool from the leaf essential oil of Cinnamomum osmophloeum Kanehira in endotoxin-induced mice[J].J Food Drug Anal,2018,26(1):211-220. doi: 10.1016/j.jfda.2017.03.006
39	ZHANGK,HUANGQ,DENGS,et al.Mechanisms of TLR4-mediated autophagy and nitroxidative stress[J].Front Cell Infect Microbiol,2021,11,766590. doi: 10.3389/fcimb.2021.766590
40	RADIR.Oxygen radicals, nitric oxide, and peroxynitrite: redox pathways in molecular medicine[J].Proc Natl Acad Sci U S A,2018,115(23):5839-5848. doi: 10.1073/pnas.1804932115
41	LIS T,DAIQ,ZHANGS X,et al.Ulinastatin attenuates LPS-induced inflammation in mouse macrophage RAW264.7 cells by inhibiting the JNK/NF-kappaB signaling pathway and activating the PI3K/Akt/Nrf2 pathway[J].Acta Pharm Sin B,2018,39(8):1294-1304. doi: 10.1038/aps.2017.143
42	崔乔. 肉桂醛对育肥绵羊生长性能、消化代谢及血液指标的影响[D]. 晋中: 山西农业大学, 2021.
	CUI Q. Effects of diet supplemented with cinnamaldehyde on growth performance, digestive metabolism and blood indicatorsin fattening lambs[D]. Jinzhong: Shanxi Agricultural University, 2021. (in Chinese)
43	WANGL,HOUY,YID,et al.Beneficial roles of dietary oleum cinnamomi in alleviating intestinal injury[J].Front Biosci (Landmark Ed),2015,20(5):814-828. doi: 10.2741/4339
44	LEALK W,LEALM,BREANCINIM,et al.Essential oils and capsaicin in the diet of Jersey cows at early lactation and their positive impact on anti-inflammatory, antioxidant and immunological responses[J].Trop Anim Health Prod,2024,56(7):247. doi: 10.1007/s11250-024-04077-w
45	MOHANTYD,PADHEES,SAHOOC,et al.Integrating network pharmacology and experimental verification to decipher the multitarget pharmacological mechanism of Cinnamomum zeylanicum essential oil in treating inflammation[J].Heliyon,2024,10(2):e24120. doi: 10.1016/j.heliyon.2024.e24120
46	SCHINKA,NAUMOSKAK,KITANOVSKIZ,et al.Anti-inflammatory effects of cinnamon extract and identification of active compounds influencing the TLR2 and TLR4 signaling pathways[J].Food Funct,2018,9(11):5950-5964. doi: 10.1039/C8FO01286E
47	HOC L,LIL H,WENGY C,et al.Eucalyptus essential oils inhibit the lipopolysaccharide-induced inflammatory response in RAW264.7 macrophages through reducing MAPK and NF-kappaB pathways[J].BMC Complement Med Ther,2020,20(1):200. doi: 10.1186/s12906-020-02999-0
48	BENH A,GARGOURIM,DHIFIW,et al.Potential anti-inflammatory and antioxidant effects of Citrus aurantium essential oil against carbon tetrachloride-mediated hepatotoxicity: A biochemical, molecular and histopathological changes in adult rats[J].Environ Toxicol,2019,34(4):388-400. doi: 10.1002/tox.22693
49	LINW T,HEY H,LOY H,et al.Essential oil from Glossogyne tenuifolia inhibits lipopolysaccharide-induced inflammation-associated genes in macro-phage cells via suppression of NF-kappaB signaling pathway[J].Plants (Basel),2023,12(6):1241.

精油 EOs	浓度/(mg·mL^-1) Concentration				SEM	P值P-value
精油 EOs	6.25	12.5	25	50	SEM	P_E	P_C	P_E*P_C	P_L	P_Q
肉桂精油CEO	57.35^a	63.87^a	74.01^a	97.95^a	0.047 1	< 0.01	< 0.01	< 0.01	< 0.01	0.60
肉桂醛CIN	23.33^b	34.16^b	46.17^b	75.75^b	0.059 2				< 0.01	0.79

精油 EOs	浓度/(mg·mL^-1) Concentration				SEM	P值P-value
精油 EOs	6.25	12.5	25	50	SEM	P_E	P_C	P_E*P_C	P_L	P_Q
肉桂精油CEO	49.20^a	77.43^a	79.73^a	84.98^a	0.0421	< 0.01	< 0.01	< 0.01	< 0.01	< 0.01
肉桂醛CIN	34.45^b	48.39^b	64.55^b	72.80^b	0.045				< 0.01	< 0.01

[1]	刘玲, 史万玉, 李秀梅, 王明华, 翟向和, 周炜炜. 陈皮精油对脂多糖诱导的RAW264.7细胞炎症的干预作用[J]. 畜牧兽医学报, 2024, 55(9): 4153-4160.
[2]	刘旭, 潘寅川, 杨亚军, 刘希望, 马宁, 李剑勇. 阿司匹林丁香酚酯对体外脂多糖诱导小鼠巨噬细胞炎症反应的抑制效应[J]. 畜牧兽医学报, 2022, 53(11): 4048-4057.
[3]	王万林, 高月, 廖加美, 彭俊超, 蔡亚玲, 易琼, 王鲁. 艾纳香油对NF-κB及Nrf2/HO-1信号通路的作用研究[J]. 畜牧兽医学报, 2021, 52(4): 976-986.
[4]	陈楷文, 华承薇, 袁宸, 潘飞, 蔺辉星, 范红结, 马喆. 马链球菌兽疫亚种烯醇化酶对小鼠肺泡巨噬细胞吞噬功能的影响[J]. 畜牧兽医学报, 2020, 51(10): 2576-2583.
[5]	贺双江, 宋瑞龙, 曹莹, 刘庆羊, 张闯, 刘宗平. 低水平镉暴露对破骨细胞分化的影响[J]. 畜牧兽医学报, 2019, 50(3): 663-669.