Dark red tongue color formation caused by hyperglycemia is attributed to decreased blood flow of tongue tissue partially due to nuclear factor-kappa B pathway activation

doi:10.19852/j.cnki.jtcm.20231018.003

Abstract

Abstract:

OBJECTIVE: To investigate the potential mechanisms underlying the dark red tongue color formation induced by hyperglycemia.
METHODS: A high-fat diet and intraperitoneal injection of streptozotocin were used to establish a diabetes model. The color and blood flow of tongues were analyzed by the Tongue Diagnosis Analysis System and laser Doppler flowmetry, respectively. Inflammatory factors and adhesion factors were measured in the circulation and tongue tissue by an enzyme-linked immunosorbent assay. Western blotting was employed to evaluate nuclear factor-kappa B (NF-κB) p50 and inhibitor of kappa B kinase protein expression levels in the tongue. Then, the NF-κB inhibitor, pyrrolidine dithiocarbamic acid ammonium salt was utilized to repress NF-κB pathway activation to validate that the NF-κB pathway plays a key role in blood flow and dark red tongue color formation.
RESULTS: The diabetic rats displayed a dark red tongue color that was accompanied by NF-κB pathway activation and decreased blood flow in the tongue. These effects could be reversed by the NF-κB inhibitor.
CONCLUSIONS: Our investigation demonstrated that hyperglycemia led to dark red tongue color formation by decreasing blood flow in the tongue, which was partly due to NF-κB pathway activation.

Key words: tongue diagnosis, type 2 diabetes mellitus, hyperglycemia, NF-kappa B, blood circulation

JIN Shenyi, LIU Yahua, HAN Xu, CAI Mengjie, XU Jiatuo, LU Hao, CHEN Qingguang. Dark red tongue color formation caused by hyperglycemia is attributed to decreased blood flow of tongue tissue partially due to nuclear factor-kappa B pathway activation[J]. Journal of Traditional Chinese Medicine, 2023, 43(6): 1118-1125.

Figures/Tables 4

Figure 1 Metabolic parameters in three groups A: FPG; B: TC; C: TG; D: LDL; E: HDL. CON: control group fed a normal diet for 8 weeks; MOD: model group fed a high-fat diet for 8 weeks; INH: NF-κB inhibitor treatment group fed a high-fat diet and administered with 200 mg/kg PDTC once daily for 8 weeks. FPG: fasting plasma glucose; TC: total cholesterol; TG: triglycerides; LDL: low-density lipoprotein; HDL: high-density lipoprotein; NF-κB: nuclear factor-kappa B; PDTC: pyrrolidine dithiocarbamic acid ammonium salt. The significant differences were examined by using a one-way analysis of variance. Data were expressed as mean ± standard deviation (n = 5-10 per group). Significant differences compared with CON were designated as aP < 0.05 and with MOD as bP < 0.05.

Figure 2 Expression of inflammatory factors in both serum and tongue of diabetic rats A-D: TNF-α, IL-1β, ICAM-1, and MCP-1 levels in tongue tissues. E-H: TNF-α, IL-1β, ICAM-1, and MCP-1 levels in serum. CON: control group fed a normal diet for 8 weeks; MOD: model group fed a high-fat diet for 8 weeks; INH: NF-κB inhibitor treatment group fed a high-fat diet and administered with 200 mg/kg PDTC once daily for 8 weeks. TNF-α: tumor necrosis factor factor-α; IL-1β: interleukin-1β; ICAM-1: intercellular adhesion molecule-1; MCP-1: monocyte chemotactic protein 1; NF-κB: nuclear factor-kappa B; PDTC: pyrrolidine dithiocarbamic acid ammonium salt. The significant differences were examined by using a one-way analysis of variance. Values are expressed as mean ± standard deviation (n = 6-10). Significant differences compared with CON were designated as aP < 0.05 and with MOD as bP < 0.05.

Table 1 Comparison of luminosity (L), a, and b color space values of tongue color in the three groups

Item	CON (n = 5-10)	MOD (n = 5-10)	INH (n = 5-10)
L	101.8±5.8	89.2±3.9^a	92.1±4.8
a	19.9±2.2	8.7±1.8^a	24.7±1.4^b
b	1.2±3.0	1.2±2.5	0.5±2.2

Figure 3 Protein levels of NF-κB p50 and IKKβ expression A: NF-κB p50 and IKKβ protein expression; B: NF-κB p50 protein level in three groups; C: IKKβ protein level in three groups. CON: control group fed a normal diet for 8 weeks; MOD: model group fed a high-fat diet for 8 weeks; INH: NF-κB inhibitor treatment group fed a high-fat diet and administered with 200 mg/kg PDTC once daily for 8 weeks. NF-κB: nuclear factor-kappa B; IKKβ: inhibitor of kappa B kinase; PDTC: pyrrolidine dithiocarbamic acid ammonium salt. The significant differences were examined by using a one-way analysis of variance. Values are expressed as mean ± standard deviation (n = 2). Significant differences compared with CON were designated as aP < 0.05 and with MOD as bP < 0.05.

References 47.

1.	International Diabetes Federation. IDF Diabetes Atlas, 10th ed. Brussels, Belgium: International Diabetes Federation 2021; 172: 108630.
2.	Ogurtsova K, Guariguata L, Barengo NC, et al. IDF diabetes Atlas: Global estimates of undiagnosed diabetes in adults for 2021. Diabetes Res Clin Pract 2022; 183: 109118. DOI URL
3.	Li J, Yuan P, Hu X, et al. A tongue features fusion approach to predicting prediabetes and diabetes with machine learning. J Biomed Inform 2021; 115: 103693. DOI URL
4.	Hu M, Lan K, Fang W, et al. Automated tongue diagnosis on the smartphone and its applications. Comput Methods Programs Biomed 2019; 174: 51-64. DOI URL
5.	Jiao W, Hu X, Tu LP, et al. Tongue color clustering and visual application based on 2D information. Int J Comput Assist Radiol Surg 2020; 15: 203-12. DOI PMID
6.	Kanawong R, Obafemi-Ajayi T, Liu D, Zhang M, Xu D, Duan Y. Tongue image analysis and its mobile app development for health diagnosis. Adv Exp Med Biol 2017; 1005: 99-121. DOI PMID
7.	Li J, Yuan P, Hu X, et al. A tongue features fusion approach to predicting prediabetes and diabetes with machine learning. J Biomed Inform 2021; 115: 103693. DOI URL
8.	Tania MH, Lwin K, Hossain MA. Advances in automated tongue diagnosis techniques. Integr Med Res 2019; 8: 42-56. DOI PMID
9.	Chen QG, Gao Y, Cai MJ, et al. A study on the objective characteristics of tongue images in the elderly with different glucose metabolism. Geriatr Health Care 2020; 26: 383-6.
10.	Hsu PC, Wu HK, Huang YC, et al. The tongue features associated with type 2 diabetes mellitus. Medicine (Baltimore) 2019; 98: e15567. DOI URL
11.	Hsu PC, Huang YC, Chiang JY, Chang HH, Liao PY, Lo LC. The association between arterial stiffness and tongue manifestations of blood stasis in patients with type 2diabetes. BMC Complement Altern Med 2016; 16: 324. DOI URL
12.	Park YJ, Yang DH, Lee JM, Park YB. Development of a valid and reliable blood stasis questionnaire and its relationship to heart rate variability. Complement Ther Med 2013; 21: 633-40. DOI URL
13.	Zheng XY. Guiding principle of clinical research on new drugs of Chinese medicine. Beijing: Chinese Medical Science and Technology Press, 2002: 29-31.
14.	Xu M, Chen H, Shi ZX, et al. Pathological observation of blood stasis syndrome in non-diabetic peripheral neuropathies: a retrospective analysis based on nerve biopsy. Chin J Integr Med 2020; 26: 776-82. DOI
15.	Liu WL, Zhang HY, Che NC, Tang ZQ, Gao LY. Changes of color and blood flow of the tongue in the mini-swine of immune hepatic injury. J Tradit Chin Med 2010; 30: 265-71. DOI URL
16.	Wang ZC, Zhang SP, Yuen PC, et al. Intra-rater and inter-rater reliability of tongue coating diagnosis in Traditional Chinese Medicine using smartphones: quasi-Delphi study. JMIR Mhealth Uhealth 2020; 8: e16018. DOI URL
17.	López-Díez R, Shen X, Daffu G, et al. Ager deletion enhances ischemic muscle inflammation, angiogenesis, and blood flow recovery in diabetic mice. Arterioscler Thromb Vasc Biol 2017; 37: 1536-47.
18.	Liu M, Li F, Liang M, Rao X. Effects of aldose reductase inhibitors on renal blood flow parameters in patients with early diabetic nephropathy. J Diabetes Complications 2020; 34: 107620. DOI URL
19.	Nagaoka T, Sato E, Takahashi A, Yokota H, Sogawa K, Yoshida A. Impaired retinal circulation in patients with type 2 diabetes mellitus: retinal laser Doppler velocimetry study. Invest Ophthalmol Vis Sci 2010; 51: 6729-34. DOI URL
20.	Betsy BD. The pathophysiology of cardiovascular disease and diabetes: beyond blood pressure and lipids. Diabetes Spectr 2008; 21: 160-5. DOI URL
21.	Jansen JF, van Bussel FC, van de Haar HJ, et al. Cerebral blood flow, blood supply, and cognition in type 2 diabetes mellitus. Sci Rep 2016; 6: 10. DOI PMID
22.	Shin YK, Hsieh YS, Kwon S, Lee HS, Seol GH. Linalyl acetate restores endothelial dysfunction and hemodynamic alterations in diabetic rats exposed to chronic immobilization stress. J Appl Physiol (1985) 2018; 124: 1274-83. DOI URL
23.	Li H, Shi Y, Wang X, et al. Piceatannol alleviates inflammation and oxidative stress via modulation of the Nrf2/HO-1 and NF-κB pathways in diabetic cardiomyopathy. Chem Biol Interact 2019; 310: 108754.
24.	Fernández J, Clària J, Amorós A, et al. Effects of albumin treatment on systemic and portal hemodynamics and systemic inflammation in patients with decompensated cirrhosis. Gastroenterology 2019; 157: 149-62.
25.	Yao L, Bhatta A, Xu Z, et al. Obesity-induced vascular inflammation involves elevated arginase activity. Am J Physiol Regul Integr Comp Physiol 2017; 313: R560-71. DOI URL
26.	Catrysse L, van Loo G. Inflammation and the metabolic syndrome: the tissue-specific functions of NF-κB. Trends Cell Biol 2017; 27: 417-29. DOI PMID
27.	Patel S, Santani D. Role of NF-κB in the pathogenesis of diabetes and its associated complications. Pharmacol. Rep 2009; 61: 595-603. DOI URL
28.	Hu R, Wang MQ, Ni SH, et al. Salidroside ameliorates endothelial inflammation and oxidative stress by regulating the AMPK/NF-κB/NLRP 3 signaling pathway in AGEs-induced HUVECs. Eur J Pharmacol 2020; 867: 172797.
29.	Liu R, Jiang LJ, Yang Y, et al. Study on syndrome differentiation strategy of phlegm and blood stasis syndromes of coronary heart disease based on expert consultation on medical cases. Ann Palliat Med 2021; 10: 9940-52. DOI PMID
30.	Cvijanovic N, Feinle-Bisset C, Young RL, Little TJ. Oral and intestinal sweet and fat tasting: impact of receptor polymorphisms and dietary modulation for metabolic disease. Nutr Rev 2015; 73: 318-34. DOI PMID
31.	Ponte E, Tabaj D, Maglione M, Melato M. Diabetes mellitus and oral disease. Acta Diabetol 2001; 38: 57-62. PMID
32.	Paneni F, Beckman JA, Creager MA, Cosentino F. Diabetes and vascular disease: pathophysiology, clinical consequences, and medical therapy: part I. Eur Heart J 2013; 34: 2436-43. DOI PMID
33.	Hartge MM, Unger T, Kintscher U. The endothelium and vascular inflammation in diabetes. Diab Vasc Dis Res 2007; 4: 84-8. DOI URL
34.	Jaap AJ, Pym CA, Seamark C, Shore AC, Tooke JE. Microvascular function in type 2 (non-insulin-dependent) diabetes: improved vasodilation after one year of good glycaemic control. Diabet Med 1995; 12: 1086-91. DOI URL
35.	Muller WA. New mechanisms and pathways for monocyte recruitment. J Exp Med 2001; 194: F47-51. DOI PMID
36.	Niu J, Kolattukudy PE. Role of MCP-1 in cardiovascular disease: molecular mechanisms and clinical implications. Clin Sci (Lond) 2009; 117: 95-109. DOI PMID
37.	Yang J, Park Y, Zhang H, et al. Role of MCP-1 in tumor necrosis factor-alpha-induced endothelial dysfunction in type 2 diabetic mice. Am J Physiol Heart Circ Physiol 2009;297: H1208-16.
38.	Alexandraki K, Piperi C, Kalofoutis C, Singh J, Alaveras A, Kalofoutis A. Inflammatory process in type 2 diabetes: the role of cytokines. Ann N Y Acad Sci 2006; 1084: 89-117. DOI URL
39.	Peiró C, Lorenzo Ó, Carraro R, Sánchez-Ferrer CF. IL-1β inhibition in cardiovascular complications associated to diabetes mellitus. Front Pharmacol 2017; 8: 363.
40.	Fischer TA. Pharmacological therapy of age-related macular degeneration based on etiopathogenesis. Orv Hetil 2015; 156: 1847-58. DOI PMID
41.	Bilsborough W, O'Driscoll G, Stanton K, et al. Effect of lowering tumour necrosis factor-alpha on vascular endothelial function in Type Ⅱ diabetes. Clin Sci (Lond) 2002; 103: 163-9. DOI URL
42.	Kracht M, Müller-Ladner U, Schmitz ML. Mutual regulation of metabolic processes and proinflammatory NF-κB signaling. J Allergy Clin Immunol 2020; 146: 694-705.
43.	Catrysse L, van Loo G. Inflammation and the metabolic syndrome: the tissue-specific functions of NF-κB. Trends Cell Biol 2017; 27: 417-29. DOI PMID
44.	Sun SC. The non-canonical NF-κB pathway in immunity and inflammation. Nat Rev Immunol 2017; 17: 545-58. DOI URL
45.	Zhang Q, Lenardo MJ, Baltimore D. 30 years of NF-κB: a blossoming of relevance to human pathobiology. Cell 2017; 168: 37-57.
46.	Bao L, Li J, Zha D, et al. Chlorogenic acid prevents diabetic nephropathy by inhibiting oxidative stress and inflammation through modulation of the Nrf2/HO-1 and NF-ĸB pathways. Int Immunopharmacol 2018; 54: 245-53.
47.	Kern TS. Contributions of inflammatory processes to the development of the early stages of diabetic retinopathy. Exp Diabetes Res 2007; 2007: 95103. DOI PMID

[1]	CHANG Fengjin, ZHOU Peng, LI Guoying, ZHANG Weizhi, ZHANG Yanyan, PENG Daiyin, CHEN Guangliang. Taohong Siwu decoction (桃红四物汤) ameliorates atherosclerosis in rats possibly through toll-like receptor 4/myeloid differentiation primary response protein 88/nuclear factor-κB signal pathway [J]. Journal of Traditional Chinese Medicine, 2024, 44(1): 103-112.
[2]	LIU Huihui, FENG Jun, LIU Jianhe, CHENG Choufu, HU Guoheng. Efficacy of Jiangzhi Xiaoban tablet (降脂消斑片) on toll-like receptor 4/nuclear factor-kappa B/nod-like receptor protein 3 signaling pathway in mice with atherosclerosis induced by high-fat diet [J]. Journal of Traditional Chinese Medicine, 2024, 44(1): 88-94.
[3]	LI He, SU Wenquan, LI Shanshan, JI Hanrui, CHENG Jiangyan, CUI Fangyuan, TANG Lu, ZHOU Li, GAO Ying, DONG Xinglu. Supplementing Qi and activating blood circulation method to treat vertebrobasilar dolichoectasia with posterior circulatory watershed infarction: a case report of two patients [J]. Journal of Traditional Chinese Medicine, 2023, 43(4): 824-828.
[4]	LUO Xin, XIE Jing, HUANG Li, GAN Wenfan, CHEN Ming. Efficacy and safety of activating blood circulation and removing blood stasis of Traditional Chinese Medicine for managing renal fibrosis in patients with chronic kidney disease: a systematic review and Meta-analysis [J]. Journal of Traditional Chinese Medicine, 2023, 43(3): 429-440.
[5]	YANG Zhangjie, WANG Yuxin, CHEN Dongmei, ZHAO Shuai, HU Na, MA Lianghong, MA Huiming. Gouqizi (Fructus Lycii) seed oil reduces D-galactose induced inflammation in testis of rats via Janus kinase 1/signal transducerand activator of transcription 1/nuclear factor-κB in vitro and in vivo [J]. Journal of Traditional Chinese Medicine, 2023, 43(2): 265-273.
[6]	XIE Jing, BI Zheng, WANG Sihai, SHEN Guoming, FANG Zhaohui. Danzhi Jiangtang capsule (丹蛭降糖胶囊) reduces renal injury in rats with diabetes induced by high fat diet and streptozotocin via downregulating toll-like receptor 4-nuclear factor-κB pathway and apoptosis [J]. Journal of Traditional Chinese Medicine, 2023, 43(2): 312-321.
[7]	ZHOU Zhenghua, JI Jianbin, WANG Hongxia, YAN Lin, KANG Hongchang. Qingchi San (青赤散) treats ulcerative colitis in mice by inhibiting the nuclear factor-kappa B signaling pathway and Nucleotide-binding oligomerization domain, leucine-rich repeat and pyrin domain-containing 3 inflammasome formation [J]. Journal of Traditional Chinese Medicine, 2023, 43(1): 68-77.
[8]	ZOU Xinshuang, SHI Lei, YIN Hailong, LI Haiping, WANG Mengheng, SONG Wanci, LUO Laichun, WU Hezhen, YANG Yanfang, ZAN Junfeng, LIU Yanwen, DAN Hanxiong, YIN Qiang, YOU Pengtao. Compound Gaoziban tablet (复方高滋斑片) alleviates depression via toll-like receptor 4/myeloid differentiation factor 88/nuclear factor-kappa B pathway [J]. Journal of Traditional Chinese Medicine, 2022, 42(6): 956-964.
[9]	XU Baogui, ZHENG Jiawen, TIAN Xiaoxiao, YUAN Falei, LIU Zhongliang, YANG Zuisu, DING Xianjun. Antihepatofibrotic effect of Guizhifuling pill (桂枝茯苓丸) on carbon tetrachloride-induced liver fibrosis in mice [J]. Journal of Traditional Chinese Medicine, 2022, 42(5): 715-722.
[10]	HE Xuemei, XIAO Junjie, FAN Chunlin, LU Zibin, CAO Huihui, YU Linzhong, ZHENG Yuanru, LIU Junshan. Zebrafish facilitates drug screening: potential of 3-deoxy-andrographoside from Chuanxinlian (Herba Andrographitis Paniculatae) as an anti-inflammatory agent [J]. Journal of Traditional Chinese Medicine, 2022, 42(5): 749-757.
[11]	ZHANG Zhaohua, LIU Rong, DU Nana, ZHU Xiangdong. Efficacy of Sishen Wan (四神丸) on dinitrobenzene sulfonic acid-induced ulcerative colitis and its effect on toll-like receptor 2/interleukin-1 receptor-associated kinase-4/nuclear factor-κB signal pathway [J]. Journal of Traditional Chinese Medicine, 2022, 42(4): 565-575.
[12]	ZHOU Ziyi, WAN Can, ZHAO Yuanqi, LIU Xiangzhe, GAO Ying, AN Hongwei, LI Lejun, ZHANG Huili, YU Xiaofei, ZHANG Xinchun, WANG Huijuan, SHI Qing, WEI Chunhua, CHEN Jie, HUANG Wenguo, CHEN Junbin, GAO Ying, HU Mingzhe, CAI Yefeng. Efficacy and safety of a sequential treatment with clearing heat and eliminating phlegm and tonifying Qi and activating blood circulation in treating acute ischemic stroke: study protocol for a randomized controlled trial [J]. Journal of Traditional Chinese Medicine, 2022, 42(4): 604-610.
[13]	Jing QIAN, Xiaohong WANG, Benzhong WEI, Guoxiong ZHOU, Shunxing ZHU, Chun LIU. Therapeutic effects of salidroside vs pyrrolidine dithiocarbamate against severe acute pancreatitis in rats [J]. Journal of Traditional Chinese Medicine, 2022, 42(1): 49-57.
[14]	CHEN Tzufan, CHEN Chuntzu, HUANG Yuanli, BASKARAN Rathinasamy, TSAI Jeffrey J.P., HU Rouhmei. Ethanolic extract of Puhuang(Pollen Typhae) modulates lipopolysaccharide-induced inflammatory response through inducible nitric oxide synthase/cyclooxygenase-2 signaling in RAW 264.7 macrophages [J]. Journal of Traditional Chinese Medicine, 2021, 41(6): 836-844.
[15]	WU Mingyun, YU Jianer, BAI Li, XUE Zheng, JIANG Shenhua, LI Liqing, PIAO Xiang, XU Wanchao, WANG Jiani, SHEN Qian. Pingchuan formula(平喘方) improves allergic asthma in mice through inhibiting nuclear factor-kappa B/mitogen-activated protein kinase signaling pathway [J]. Journal of Traditional Chinese Medicine, 2021, 41(6): 883-890.

Home

Online First

Author Center

Reviewer Center

Issues

Announcement

About Us