小檗碱联合姜黄素调控M1巨噬细胞极化对动脉粥样硬化的作用机制

doi:10.3969/j.issn.1006-5725.2024.14.003

Abstract

Abstract:

Objective To explore the effect and mechanism of berberine combined with curcumin on atherosclerosis （AS） by mediating M1 macrophages polarization. Methods M1-type macrophages were obtained from mouse mononuclear macrophages （RAW264.7） induced by lipopolysaccharide （LPS， 100 ng/mL） and interferon （IFN）-γ （20 ng/mL）. A cell model was established. The cells were divided into a control group， model group， berberine group， curcumin group and berberine plus curcumin group. Concentrations of berberine and curcumin were detected by CCK-8 assay. The expression levels of M1-type macrophage markers iNOS， TNF-α， CXCL9 and p-STAT6/STAT6 in macrophage supernatant were detected by ELISA. Levels of iNOS， TNF-α and CXCL9 mRNA were detected by RT-PCR. Levels of iNOS， STAT6 and p-STAT6 proteins in each group were detected by Western blot. After down-regulation of STAT6 level by siRNA technology， expression of p-STAT6 protein was detected by Western blot. Expression levels of iNOS， TNF-α， CXCL9 and p-STAT6 were detected by ELISA. Results In the polarization of M1 macrophages induced by LPS and IFN-γ， berberine （25 μmol/L） and curcumin （20 μmol/L） were the best concentrations as compared with other drug concentration groups， and neither alone nor combined use could significantly inhibit the viability of RAW264.7 cells （P < 0.05）. As compared with the normal group， iNOS， TNF-α and CXCL9 mRNA and protein levels were increased in the model group， while P-STAT6/STAT6 levels were decreased， with statistical differences （P < 0.05）. As compared with the model group， iNOS， TNF-α and CXCL9 mRNA and protein levels in the berberine group， curcumin group， and berberine plus curcumin group were decreased， while P-STAT6 /STAT6 levels were increased， and the changes were more obvious in berberine plus curcumin group， with statistical difference （P < 0.05）. After transfection of STAT6 siRNA in M1 macrophages in the berberine plus curcumin group， P-STAT6 levels were down-regulated， while expressions of iNOS， TNF-α and CXCL9 were up-regulated， with statistical differences （P < 0.05）. Conclusions Both berberine and curcumin can inhibit the activity of M1-type macrophages and reduce inflammatory response. The action of berberine combined with curcumin is more advantageous than that of either drug alone， which may be the main mechanism of action through activation of STAT6.

Key words: atherosclerosis, M1 macrophage, berberine, curcumin, STAT6

CLC Number:

R543.5

Yushan CHEN,Tingting WANG,Xinyi HAN,Chengjun HUA,Boyuan JIN,Shasha SHANG,Yonghua ZONG,Yazhou. LIANG. Effect of M1 macrophage polarization regulated by berberine combined with curcumin on atherosclerosis[J]. The Journal of Practical Medicine, 2024, 40(14): 1915-1921.

Figures/Tables 6

Tab.1

Fig.1

Fig.2

Fig.3

Fig.4

Fig.5

References 26

1	BLAGOV A V， MARKIN A M， BOGATYREVA A I， et al. The role of macrophages in the pathogenesis of atherosclerosis［J］. Cells， 2023， 12（4）： 522. doi:10.3390/cells12040522 doi: 10.3390/cells12040522
2	HOU P， FANG J， LIU Z， et al. Macrophage polarization and metabolism in atherosclerosis［J］. Cell Death Dis， 2023， 14（10）： 691. doi:10.1038/s41419-023-06206-z doi: 10.1038/s41419-023-06206-z
3	HE Y， LIU T. Oxidized low-density lipoprotein regulates macrophage polarization in atherosclerosis［J］. Int Immunopharmacol， 2023， 120： 110338. doi:10.1016/j.intimp.2023.110338 doi: 10.1016/j.intimp.2023.110338
4	HE P， WANG H， CHENG S， et al. Geniposide ameliorates atherosclerosis by regulating macrophage polarization via perivascular adipocyte-derived CXCL14［J］. J Ethnopharmacol， 2023， 314： 116532. doi:10.1016/j.jep.2023.116532 doi: 10.1016/j.jep.2023.116532
5	ZHOU L， ZHAO H， ZHAO H， et al. GBP5 exacerbates rosacea‐like skin inflammation by skewing macrophage polarization towards M1 phenotype through the NF‐κB signalling pathway［J］. J Eur Acad Dermatol Venereol， 2023， 37（4）： 796-809. doi:10.1111/jdv.18725 doi: 10.1111/jdv.18725
6	HSU C H， PAN Y J， ZHENG Y T， et al. Ultrasound reduces inflammation by modulating M1/M2 polarization of microglia through STAT1/STAT6/PPARγ signaling pathways［J］. CNS Neurosci Ther， 2023， 29（12）： 4113-4123. doi:10.1111/cns.14333 doi: 10.1111/cns.14333
7	GUO H H， SHEN H R， WANG L L， et al. Berberine is a potential alternative for metformin with good regulatory effect on lipids in treating metabolic diseases［J］. Biomed Pharmacother， 2023， 163： 114754. doi:10.1016/j.biopha.2023.114754 doi: 10.1016/j.biopha.2023.114754
8	LAURINDO L F， DE CARBALHO G M， DE OLIVERIRA Zanuso B， et al. Curcumin-Based Nanomedicines in the Treatment of Inflammatory and Immunomodulated Diseases： An Evidence-Based Comprehensive Review［J］. Pharmaceutics， 2023， 15（1）： 229. doi:10.3390/pharmaceutics15010229 doi: 10.3390/pharmaceutics15010229
9	XIONG K， DENG J， YUE T， et al. Berberine promotes M2 macrophage polarisation through the IL-4-STAT6 signalling pathway in ulcerative colitis treatment［J］. Heliyon， 2023，9（3）：e14176. doi:10.1016/j.heliyon.2023.e14176 doi: 10.1016/j.heliyon.2023.e14176
10	ABDOLLAHI E， JOHNSTON TP， GHANEIFAR Z， et al. Immunomodulatory Therapeutic Effects of Curcumin on M1/M2 Macrophage Polarization in Inflammatory Diseases［J］. Curr Mol Pharmacol， 2023，16（1）：2-14. doi:10.2174/1874467215666220324114624 doi: 10.2174/1874467215666220324114624
11	黄勉，李琳，李芬，等. 小檗碱对LPS诱导神经元损伤的作用机制研究［J］. 现代免疫学， 2023，43（6）：465-472.
12	CAO L， YUAN X， ZHANG F E， et al. Effects of curcumin on proliferation and apoptosis and migration of human pterygium fibroblasts［J］. Int Eye Science， 2023： 731-737.
13	HUANG Z， SHEN S， HAN X， et al. Macrophage DCLK1 promotes atherosclerosis via binding to IKKβ and inducing inflammatory responses［J］. EMBO Mol Med， 2023， 15（5）： e17198. doi:10.15252/emmm.202217198 doi: 10.15252/emmm.202217198
14	HUANG H， SUN Z， XU J， et al. Yang-Xin-Shu-Mai granule alleviates atherosclerosis by regulating macrophage polarization via the TLR9/MyD88/NF-κB signaling pathway［J］. J Ethnopharmacol， 2024， 318： 116868. doi:10.1016/j.jep.2023.116868 doi: 10.1016/j.jep.2023.116868
15	JINNOUCHI H， GUO L， SAKAMOTO A， et al. Diversity of macrophage phenotypes and responses in atherosclerosis ［J］. Cell Mol Life Sci， 2020， 77（10）： 1919-1932. doi:10.1007/s00018-019-03371-3 doi: 10.1007/s00018-019-03371-3
16	BARRETT TJ. Macrophages in Atherosclerosis Regression ［J］. Arterioscler Thromb Vasc Biol， 2020， 40（1）： 20-33. doi:10.1161/atvbaha.119.312802 doi: 10.1161/atvbaha.119.312802
17	WEI J， SHEN S， TIAN Y， et al. Correlation analysis of macrophage distribution and pathological features of carotid atherosclerotic plaque［J］. Ann Vasc Surg， 2024，98：355-364. doi:10.1016/j.avsg.2023.08.030 doi: 10.1016/j.avsg.2023.08.030
18	JIAN X， LIU Y， ZHAO Z， et al. The role of traditional Chinese medicine in the treatment of atherosclerosis through the regulation of macrophage activity［J］. Biomed Pharmacother， 2019， 118： 109375. doi:10.1016/j.biopha.2019.109375 doi: 10.1016/j.biopha.2019.109375
19	ADEL-MEHRABAN M S， AGHABEIGLOOEI Z， ATLASI R， et al. Berberine as a Natural Modifier of Gut Microbiota to Promote Metabolic Status in Animal Studies and Clinical Trials： A Systematic Review［J］. Trad Integrat Med， 2023， 2023：202-216. doi:10.18502/tim.v8i2.13086 doi: 10.18502/tim.v8i2.13086
20	DASH R， YADAV M， BISWAL J， et al. Modeling of chitosan modified PLGA atorvastatin-curcumin conjugate （AT-CU） nanoparticles， overcoming the barriers associated with PLGA： An approach for better management of atherosclerosis［J］. Int J Pharm， 2023， 640： 123009. doi:10.1016/j.ijpharm.2023.123009 doi: 10.1016/j.ijpharm.2023.123009
21	LI Z H， CHEN J F， ZHANG J， et al. Mesenchymal stem cells promote polarization of M2 macrophages in mice with acute-on-chronic liver failure via Mertk/JAK1/STAT6 signaling［J］. Stem Cells， 2023， 41（12）： 1171-1184. doi:10.1093/stmcls/sxad069 doi: 10.1093/stmcls/sxad069
22	GERASIMOVA E V， POPKOVA T V， GERASIMOVA D A， et al. Macrophage dysfunction in autoimmune rheumatic diseases and atherosclerosis［J］. Int J of Mol Sci， 2022， 23（9）： 4513. doi:10.3390/ijms23094513 doi: 10.3390/ijms23094513
23	YANG T， WANG R， LIU H， et al. Berberine regulates macrophage polarization through IL-4-STAT6 signaling pathway in Helicobacter pylori-induced chronic atrophic gastritis ［J］. Life Sci， 2021， 266： 118903. doi:10.1016/j.lfs.2020.118903 doi: 10.1016/j.lfs.2020.118903
24	JIANG M， QI Y， HUANG W， et al. Curcumin reprograms TAMs from a protumor phenotype towards an antitumor phenotype via inhibiting MAO-A/STAT6 pathway［J］. Cells， 2022， 11（21）： 3473. doi:10.3390/cells11213473 doi: 10.3390/cells11213473
25	XIONG K， DENG J， YUE T， et al. Berberine promotes M2 macrophage polarisation through the IL-4-STAT6 signalling pathway in ulcerative colitis treatment［J］. Heliyon， 2023，9（3）：e14176. doi:10.1016/j.heliyon.2023.e14176 doi: 10.1016/j.heliyon.2023.e14176
26	AMIRI Z， JALILI S， TARAHOMI M， et al. Curcumin′s spice-infused therapeutic promise： disease severity alleviation in a mouse model of multiple sclerosis via modulation of immune responses［J］. Mol Biol Rep， 2023， 50（11）： 8843-8853. doi:10.1007/s11033-023-08781-y doi: 10.1007/s11033-023-08781-y

目的基因	上游引物序列（5′?3′）	下游引物序列（5′?3′）	扩增片段（bp）
iNOS	CACCACCCTCCTCGTTC	CAATCCACAACTCGCTCC	132
TNF-α	ACGGCATGGATCTCAAAGAC	AGATAGCAAATCGGCTGACG	138
CXCL9	CCGAGGCACGATCCACTACA	CCGGATCTAGGCAGGTTTGA	122
GAPDH	CCTGGAGAAACCTGCCAAGTA	TCATACCAGGAAATGAGCTTGAC	201

[1]	Jia CUI,Mingming ZHANG,Na WANG,Wei GAO,Fei LI. Correlation analysis of glucose and lipid metabolism indicators combined with antiphospholipid antibodies on prognosis of patients with atherosclerosis [J]. The Journal of Practical Medicine, 2024, 40(3): 326-329.
[2]	Tingting WANG,Lili YU,Xiangli SHEN,Junmeng ZHENG,Yushan CHEN,Shasha SHANG,Jianru. WANG. Application and research progress of nanotechnology in atherosclerosis [J]. The Journal of Practical Medicine, 2024, 40(1): 53-58.
[3]	. Amygdalin reduces coronary endothelial pyroptosis and ameliorates atherosclerotic plaque formation in ApoE-/- mice [J]. The Journal of Practical Medicine, 2023, 39(14): 1746-1755.
[4]	QIN Hewei, NIU Yuqing, SONG Xuemei, WANG Mengnan, SUN Mengyan.. Molecular mechanism of Xueguan Ruanhua Pill in preventing atherosclerosis through inhibiting vascular inflammation [J]. The Journal of Practical Medicine, 2022, 38(15): 1901-1907.
[5]	XU Ziyuan, ZHANG Jiu′na, MENG Cancan, CHENG Yan, GUO Yongze, LI Xiaotian. . Effects of Lactobacillus rhamnosus and curcumin on MAFLD by regulating TLR4 signaling pathway [J]. The Journal of Practical Medicine, 2022, 38(13): 1624-1629.
[6]	TAO Xiao⁃ fang, WANG Yajie, WANG Wenbin, FEI Nianhua. . Regulatory role of Map4k4 protein expression in atherosclerotic vulnerable plaque formation [J]. The Journal of Practical Medicine, 2022, 38(11): 1346-1352.
[7]	WEI Hui, LI Hui, YU Guohua, DONG Sujuan, ZHU Weifang.. Effects of IL⁃38 on atherosclerosis in ApoE-/- mice and their mechanisms [J]. The Journal of Practical Medicine, 2021, 37(7): 858-862.
[8]	LIU Yang, SUN Yue, YANG Anning, LIU Zige, LIU Taiyang, HAO Wei, LIU Yaoyang, WANG Qiushi, LIU Zhihong. Involvement of ferroptosis in atherosclerosis induced by high⁃fat diet in ApoE-/- mouse and formation of ox ⁃LDL ⁃induced foam cell [J]. The Journal of Practical Medicine, 2021, 37(5): 585-590.
[9]	CHU Ye, CHENG Li, JIN Jing, LIU Jianyong.. Correlations between serum γ⁃glutamyltransferase and carotid atherosclerosis in patients with obstructive sleep apnea hypopnea syndrome [J]. The Journal of Practical Medicine, 2021, 37(20): 2651-2655.
[10]	CAO Zhiyong, LU Zhenhui, GUO Xiaoming, ZHU Xiangyang.. Effect of Ezetimibe combined with Atorvastatin on the short⁃term prognosis and recurrence of mild stroke and intracranial atherosclerosis [J]. The Journal of Practical Medicine, 2021, 37(19): 2518-2523.
[11]	ZHANG Siting, CAI Yan, LI Bi, SUN Zhidan.. Detection of carotid artery elastic function with ultrafast ultrasound imaging technology in patients with gouty arthritis [J]. The Journal of Practical Medicine, 2021, 37(19): 2535-2539.
[12]	ZHANG Yue, HUA Jia, LIU Xiaobin, ZHANG Xiran, XUE Jing, WANG Liang. Wuxi. Association of cardiac valve calcification and carotid atherosclerosis on prognosis in maintenance peritoneal dialysis patients [J]. The Journal of Practical Medicine, 2021, 37(18): 2360-2365.
[13]	FAN Xiaolin, YANG Xiaohua, ZHOU Xiao, LI Xiaofei, SU Ruting, CHEN Zhigang, LIU Hua. . Clinical efficacy of berberine in treatment of acute gouty arthritis and its effect on serum levels of inflam⁃ mation factors [J]. The Journal of Practical Medicine, 2021, 37(18): 2413-2422.
[14]	FANG Yanling, WEI Yutao, XIAO Xinhuai, XU Miqing, LAN Dan, XU Jieyi, JI⁃ANG Xingzuan, YAO Jianling.. Analysis on the factors influencing quality of life of elderly patients with type 2 diabetes mellitus and coro⁃nary arteries atherosclerosis [J]. The Journal of Practical Medicine, 2020, 36(23): 3250-3255.

Effect of M1 macrophage polarization regulated by berberine combined with curcumin on atherosclerosis

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 6

References 26

Related Articles 14

Recommended Articles

Metrics

Comments