Qingwei San Treats Oral Ulcer Subjected to Stomach Heat Syndrome in db/db Mice by Targeting TLR4/MyD88/NF-κB Pathway

Abstract Background Qingwei San (QWS), one of classic Chinese Medicine prescripts, has been widely used to treat stomach heat syndrome which manifests oral ulcer (OU), periodontitis and upper gastrointestinal bleeding for seven hundred years. However, the therapeutic effects of QWS on diabetic OU subjected to stomach heat syndrome are still ambiguous. In the study, we investigate the pharmacological mechanisms. Methods The main components of QWS aqueous extract were analyzed by LC-MS, and potential pathways of QWS targeting OU were predicted by network pharmacology. The db/db mice were administered with the decoction of dried Zingiber officinale Rosc. rhizome combined with NaOH cauterization to establish the model of diabetic OU subjected to stomach heat syndrome. Subsequently, the model mice were treated with QWS, and OU wound healing status were recorded. The pathological changes of gastric tissue and oral mucosa were evaluated using hematoxylin-eosin staining, and the morphology of collagen fibers in oral mucosa was assessed by Masson staining. The levels of thromboxane B 2 (TXB 2 ), 6-Keto-prostaglandin F1α (6-keto-PGF1α), interleukin-1 β (IL-1β), IL-2, IL-6, tumor necrosis factor-α (TNF-α), β-endorphin (β-EP) and 5-Hydroxytryptamine (5-HT) were determined by ELISA assay. The protein expressions of Toll-like receptor 4 (TLR4), TNF receptor associated factor 6 (TRAF6), myeloid differentiation factor 88 (MyD88), inhibitor of NF-κB alpha (IκΒα), p-IκΒα and nuclear factor kappa-B (NF-κB) p65 were measured by Western Blotting. Results A total of 183 compounds in QWS were identified by LC-MS, and identified 79 bioactive compounds corresponded to 269 targets and 59 pathways. QWS high-dose treatment significantly reduced the level of TXB 2 and the ratio of TXB 2 /6-keto-PGF1α. Meanwhile, it improved mucosal pathological morphology, and reduced the area of OU and local edema. Simultaneously, the levels of TNF-α, IL-1β, IL-6, IL-2 and 5-HT, and the expressions of TLR4、TRAF6、MyD88、p-IκΒα and NF-κB p65 were decreased. Conclusion QWS treatment facilitates the healing of OU, ameliorates pathological morphologies of gastric and oral mucosa and decreases the levels of pro-inflammatory cytokines in db/db mice subjected to stomach heat syndrome, whose mechanism may be associated with the inhibition of TLR4/MyD88/NF-κB signaling pathway to exert anti-inflammatory effects.

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Qingwei San treats oral ulcer subjected to stomach heat syndrome in db/db mice by targeting TLR4/MyD88/NF-κB pathway

Chinese Medicine ◽

10.1186/s13020-021-00565-5 ◽

2022 ◽

Vol 17 (1) ◽

Author(s):

Lu Shi ◽

Yongcheng An ◽

Long Cheng ◽

Yiyang Li ◽

Huimin Li ◽

...

Keyword(s):

Oral Mucosa ◽

Interleukin 1 ◽

Zingiber Officinale ◽

Oral Ulcer ◽

Network Pharmacology ◽

High Dose ◽

Therapeutic Effects ◽

Tnf Α ◽

Factor Α ◽

Heat Syndrome

Abstract Background Qingwei San (QWS), one of classic Chinese Medicine prescripts, has been widely used to treat stomach heat syndrome which manifests oral ulcer (OU), periodontitis and upper gastrointestinal bleeding for seven hundred years. However, the therapeutic effects of QWS on diabetic OU subjected to stomach heat syndrome are still ambiguous. In the study, we investigated the pharmacological mechanisms. Methods The main components of QWS aqueous extract were analyzed by LC–MS, and potential pathways of QWS targeting OU were predicted by network pharmacology. The db/db mice were administered with the decoction of dried Zingiber officinale Rosc. rhizome combined with NaOH cauterization to establish the model of diabetic OU subjected to stomach heat syndrome. Subsequently, the model mice were treated with QWS, and OU wound healing status were recorded. The pathological changes of gastric tissue and oral mucosa were evaluated using hematoxylin–eosin staining, and the morphology of collagen fibers in oral mucosa was assessed by Masson staining. The levels of thromboxane B2 (TXB2), 6-Keto-prostaglandin F1α (6-keto-PGF1α), interleukin-1 β (IL-1β), IL-2, IL-6, tumor necrosis factor-α (TNF-α), β-endorphin (β-EP) and 5-Hydroxytryptamine (5-HT) were determined by ELISA assay. The protein expressions of Toll-like receptor 4 (TLR4), TNF receptor associated factor 6 (TRAF6), myeloid differentiation factor 88 (MyD88), inhibitor of NF-κB alpha (IκΒα), p-IκΒα and nuclear factor kappa-B (NF-κB) p65 were measured by Western Blotting. Results A total of 183 compounds in QWS were identified by LC–MS, and identified 79 bioactive compounds corresponded to 269 targets and 59 pathways. QWS high-dose treatment significantly reduced the level of TXB2 and the ratio of TXB2/6-keto-PGF1α. Meanwhile, it improved mucosal pathological morphology, and reduced the area of OU and local edema. Simultaneously, the levels of TNF-α, IL-1β, IL-6, IL-2 and 5-HT, and the expressions of TLR4, TRAF6, MyD88, p-IκΒα and NF-κB p65 were decreased. Conclusion QWS treatment facilitates the healing of OU, ameliorates pathological morphologies of gastric and oral mucosa and decreases the levels of pro-inflammatory cytokines in db/db mice subjected to stomach heat syndrome, whose mechanism may be associated with the inhibition of TLR4/MyD88/NF-κB signaling pathway to exert anti-inflammatory effects.

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Simultaneous Inhibition of Three Major Cytokines and Its Therapeutic Effects: A Peptide-Based Novel Therapy against Endotoxemia in Mice

Journal of Personalized Medicine ◽

10.3390/jpm11050436 ◽

2021 ◽

Vol 11 (5) ◽

pp. 436

Author(s):

Hung-Jen Shih ◽

Chao-Yuan Chang ◽

Milton Chiang ◽

Van Long Le ◽

Hao-Jen Hsu ◽

...

Keyword(s):

Liver Injury ◽

Therapeutic Effects ◽

The Novel ◽

Tissue Water ◽

Novel Therapy ◽

Binding Domains ◽

Tnf Α ◽

Simultaneous Inhibition ◽

Factor Α ◽

Liver Tissues

Three major cytokines, including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and IL-6, mediate endotoxemia-induced liver injury. With the similar structures to the binding domains of the three cytokines to their cognate receptors, the novel peptide KCF18 can simultaneously inhibit TNF-α, IL-1β, and IL-6. We elucidated whether KCF18 can alleviate injury of liver in endotoxemic mice. Adult male mice (BALB/cJ) were intraperitoneally (i.p.) administered lipopolysaccharide (LPS, 15 mg/kg; LPS group) or LPS with KCF18 (LKCF group). Mice in the LKCF group received KCF18 (i.p.) at 2 h (0.6 mg/kg), 4 h (0.3 mg/kg), 6 h (0.3 mg/kg), and 8 h (0.3mg/kg) after LPS administration. Mice were sacrificed after receiving LPS for 24 h. Our results indicated that the binding levels of the three cytokines to their cognate receptors in liver tissues in the LKCF group were significantly lower than those in the LPS group (all p < 0.05). The liver injury level, as measured by performing functional and histological analyses and by determining the tissue water content and vascular permeability (all p < 0.05), was significantly lower in the LKCF group than in the LPS group. Similarly, the levels of inflammation (macrophage activation, cytokine upregulation, and leukocyte infiltration), oxidation, necroptosis, pyroptosis, and apoptosis (all p < 0.05) in liver tissues in the LKCF group were significantly lower than those in the LPS group. In conclusion, the KCF18 peptide–based simultaneous inhibition of TNF-α, IL-1β, and IL-6 can alleviate liver injury in mice with endotoxemia.

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Tumor necrosis factor-α and interleukin-1 β levels in recurrent and persistent otitis media with effusion

Otolaryngology ◽

10.1067/mhn.2002.124188 ◽

2002 ◽

Vol 126 (4) ◽

pp. 417-422 ◽

Cited By ~ 7

Author(s):

Sertac Yetiser ◽

Bulent Satar ◽

Atilla Gumusgun ◽

Faruk Unal ◽

Yalcin Ozkaptan

Keyword(s):

Tumor Necrosis Factor ◽

Otitis Media ◽

Tumor Necrosis ◽

Otitis Media With Effusion ◽

Interleukin 1 ◽

Tertiary Referral Center ◽

Adenoid Tissue ◽

Tnf Α ◽

Factor Α ◽

Necrosis Factor

OBJECTIVE: Based on interleukin (IL)-1β and tumor necrosis factor (TNF)-α levels in effusions, our goals were to specify either recurrent or persistent otitis media with effusion (OME) is a mid stage in the development of chronic disease and to identify the factors that have an influence on cytokine levels. STUDY DESIGN: Samples from groups with recurrent (n = 15) and persistent (n = 39) OME were essayed for IL-1 β and TNF-α. Children were also grouped with respect to age, sex, quality of effusion, and the presence of pharyngeal adenoid tissue. SETTING: Tertiary referral center. RESULTS: In recurrent and persistent OME groups, IL-1β was higher than TNF-α ( P < 0.01). IL-β was higher in recurrent OME than in persistent OME ( P < 0.05). CONCLUSION: Recurrent OME seems to be closer to the chronic stage of the disease relative to persistent OME in terms of higher IL-1 β levels. Each exacerbation of acute disease in recurrent otitis media is likely to be mediated by IL-1 β. SIGNIFICANCE: We were able to clarify that recurrent OME is a stage that occurs before chronic OME. Therefore, the prevention of acute attacks in recurrent disease would also impede long-term damage to the middle ear.

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Predisposition to Hyperthyroidism May Be Influenced by Functional TNF-α, IL-1, IL-6, and IL-10 Polymorphisms: A Meta-Analysis

International Archives of Allergy and Immunology ◽

10.1159/000508284 ◽

2020 ◽

Vol 181 (12) ◽

pp. 956-965

Author(s):

Hong Ma ◽

Ting Tan ◽

Jie Wu ◽

Juan Chen ◽

Xiaohong Zhang

Keyword(s):

Meta Analysis ◽

Interleukin 1 ◽

Interleukin 10 ◽

Interleukin 4 ◽

Asian Origin ◽

Tnf Α ◽

Caucasian Origin ◽

Meta Analyses ◽

Factor Α ◽

Necrosis Factor

Background: Predisposition to hyperthyroidism may be influenced by functional gene polymorphisms in tumor necrosis factor-α (TNF-α), interleukin-1 (IL-1), interleukin-4 (IL-4), interleukin-6 (IL-6), and interleukin-10 (IL-10). However, the results of the studies published so far remain discrepant, so we conducted a meta-analysis to more robustly investigate relationships between TNF-α/IL-1/IL-4/IL-6/IL-10 polymorphisms and predisposition to hyperthyroidism. Methods: A comprehensive literature retrieval from PubMed, Embase, Web of Science, WanFang, VIP, and CNKI was endorsed by the authors, and 38 studies were found to be eligible for pooled meta-analyses. Results: We found that genotypic frequencies of TNF-α −308 G/A, IL-1A −889 C/T, IL-6 −174 G/C, IL-6 −572 G/C, IL-10 −819 C/T, and IL-10 −1082 A/G polymorphisms among cases were significantly different from those among controls. Moreover, we also found that genotypic frequencies of TNF-α −308 G/A and IL-6 −174 G/C polymorphisms among cases of Caucasian origin were significantly different from those among Caucasian controls, and genotypic frequencies of IL-1A −889 C/T, IL-1B −511 C/T, IL-6 −174 G/C, IL-6 −572 G/C, and IL-10 −1,082 A/G polymorphisms among cases of Asian origin were also significantly different from those among Asian controls. Conclusions: This meta-analysis suggests that TNF-α −308 G/A, IL-1A −889 C/T, IL-1B −511 C/T, IL-6 −174 G/C, IL-6 −572 G/C, IL-10 −819 C/T, and IL-10 −1,082 A/G polymorphisms may influence predisposition to hyperthyroidism in certain ethnic groups.

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The Augmented Productions of Neopterin and Cytokines from Macrophages/monocytes in vitro

Pteridines ◽

10.1515/pteridines.1996.7.3.72 ◽

1996 ◽

Vol 7 (3) ◽

pp. 72-76

Author(s):

Tadashi Lizuka ◽

Mitsuyo Sasaki ◽

Hitomi Kamisako ◽

Ko Oishi ◽

Shigeru Uemura ◽

...

Keyword(s):

Kawasaki Disease ◽

Interleukin 1 ◽

Poly I:C ◽

Recombinant Interferon ◽

Preliminary Examination ◽

Tnf Α ◽

Ifn Γ ◽

Factor Α ◽

Necrosis Factor

Summary In Kawasaki disease patients, increases in excretion of urinary neopterin coincided with fever and monocytosis in peripheral blood. We examined the products of neopterin, tumor necrosis factor-α (TNFα) and Interleukin-1 β (1L-1β) from healthy adult macrophages/monocytes (Mφ>/M), after stimulation with several activators to obtain some understanding of Kawasaki disease. Upon stimulation with either lipopolysaccharide (LPS) or polyinosinate-polycytidylate (Poly I:C), the Mφ/M released neopterin and pyogenic products (TNF-α or 1L-1β). The release of neopterin was eliminated by the addition of the anti-interferon-y antibody. The production of both TNF-α, 1L-1β and neopterin from Mφ/M upon stimulation of LPS was augmented in a co-culture with low dose recombinant interferon-y (rIFN-γ). Upon stimulation with rIFN-γ alone, however, the Mφ/M released neopterin but not the pyogenic products. A preliminary examination failed to detect. any difference in the response of the Mφ/M in adults annd children after stimulation with LPS. We concluded that some endotoxins could trigger the onset of Kawasaki disease and that endogenous IFN-γ can play an important role in the abnormality of Kawasaki disease patients

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Inhibition of TNF-α production contributes to the attenuation of LPS-induced hypophagia by pentoxifylline

AJP Regulatory Integrative and Comparative Physiology ◽

10.1152/ajpregu.2000.279.6.r2113 ◽

2000 ◽

Vol 279 (6) ◽

pp. R2113-R2120 ◽

Cited By ~ 26

Author(s):

M. H. Porter ◽

B. J. Hrupka ◽

G. Altreuther ◽

M. Arnold ◽

W. Langhans

Keyword(s):

Bacterial Infections ◽

Tumor Necrosis ◽

Low Dose ◽

Potent Inhibitor ◽

Muramyl Dipeptide ◽

High Dose ◽

Tnf Α ◽

Anorectic Effect ◽

Factor Α ◽

Necrosis Factor

Cytokines such as tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) are assumed to mediate anorexia during bacterial infections. To improve our understanding of the role that these two cytokines serve in mediating infection during anorexia, we investigated the ability of pentoxifylline (PTX), a potent inhibitor of TNF-α production, to block the anorectic effects of the bacterial products lipopolysaccharide (LPS) and muramyl dipeptide (MDP) in rats. Intraperitoneally injected PTX (100 mg/kg body wt) completely eliminated the anorectic effect of intraperitoneally injected LPS (100 μg/kg body wt) and attenuated the anorectic effect of a higher dose of intraperitoneally injected LPS (250 μg/kg body wt). Concurrently, PTX pretreatment suppressed low-dose LPS-induced TNF-α production by more than 95% and IL-1β production 39%, as measured by ELISA. Similarly, high-dose LPS-induced TNF-α production was reduced by ∼90%. PTX administration also attenuated the tolerance that is normally observed with a second injection of LPS. In addition, PTX pretreatment attenuated the hypophagic effect of intraperitoneally injected MDP (2 mg/kg body wt) but had no effect on the anorectic response to intraperitoneally injected recombinant human TNF-α (150 ug/kg body wt). The results suggest that suppression of TNF-α production is sufficient to attenuate LPS- and MDP-induced anorexia. This is consistent with the hypothesis that TNF-α plays a major role in the anorexia associated with bacterial infection.

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Intravenous Liposomal Prednisolone Downregulates In Situ TNF-α Production by T-cells in Experimental Autoimmune Encephalomyelitis

Journal of Histochemistry & Cytochemistry ◽

10.1177/002215540305100915 ◽

2003 ◽

Vol 51 (9) ◽

pp. 1241-1244 ◽

Cited By ~ 16

Author(s):

Jens Schmidt ◽

Josbert M. Metselaar ◽

Ralf Gold

Keyword(s):

T Cells ◽

Experimental Autoimmune Encephalomyelitis ◽

High Dose ◽

Autoimmune Encephalomyelitis ◽

Double Labeling ◽

Formalin Fixed Paraffin ◽

Tnf Α ◽

Formalin Fixed Paraffin Embedded ◽

Factor Α ◽

Experimental Autoimmune

Multiple sclerosis (MS) relapses are treated with high-dose IV glucocorticosteroids. Here we investigated mechanisms of long-circulating polyethylene glycol-coated liposomes encapsulating prednisolone (PL) in adoptive transfer experimental autoimmune encephalomyelitis. Rats received IV 10 mg/kg PL 6, 18, or 42 hr before sacrifice at disease maximum. In formalin-fixed, paraffin-embedded spinal cord we employed a nonfluorescent immunohistochemical (IHC) double labeling. We stained for tumor necrosis factor-α (TNF-α) in combination with a T-cell antigen. Compared with PBS-containing liposomes, PL at 18 hr, and more at 42 hr, significantly reduced the rate of TNF-α double-labeled T-cells. This correlated with an ameliorated disease score at day 5 after PL 42 hr. Our results help to further understand mechanisms of action of drug targeting by liposomal steroids, with possible implications for treatment of autoimmune disorders such as MS.

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Antimicrobial, Antioxidant and Anti-inflammation Activities of Fraction and Single Peptides Derived from Mare Milk Protein

Jurnal Ilmu Ternak dan Veteriner ◽

10.14334/jitv.v24i3.1976 ◽

2019 ◽

Vol 24 (3) ◽

pp. 112

Author(s):

Eni Kusumaningtyas ◽

Didik Tulus Subekti ◽

D FL Fitaningtyas

Keyword(s):

Milk Protein ◽

Animal Health ◽

Interleukin 1 ◽

Antibacterial And Antifungal Activities ◽

Tnf Α ◽

Factor Α ◽

Antibacterial And Antifungal ◽

Anti Inflammation ◽

Single Peptide ◽

Better Than

Mare milk protein contains bioactive peptide which beneficial for human and animal health. Peptides in the fraction and single may show different activities. The objectives of the study were to evaluate antimicrobial and anti-inflammation activities of the fraction and single peptide derived from mare milk protein. Antimicrobial assay was conducted by testing antibacterial and antifungal activities of fraction <3 kDa, peptide H<tt>PYFYAPELLYYANK, </tt>LVNELTEFAK and LANSLTEFAK against Escherichia coli and Candida albicans. Anti-inflammation effect was detected by interleukin 1-β (IL- 1β) and Tumor Necrosis Factor -α (TNF-α) production in mice after administration of Escherichia coli’s lipopolysaccharide (LPS) and combined with fraction or single peptide. The result showed that antibacterial and antifungal of fraction <3 kDa was higher than all of single peptide. This may because of synergistic interaction among peptide in the fraction which increase the activities. Fraction <3 kDa was also able to decrease production of IL-1β and TNF-α better than single peptide indicated its ability to decrease inflammation. Based on the results, antimicrobial and anti-inflammation activities fraction <3 kDa was better than single peptide.

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BNIP3 decreases the LPS-induced inflammation and apoptosis of chondrocytes by promoting the development of autophagy

10.21203/rs.3.rs-29090/v2 ◽

2020 ◽

Author(s):

Zetao Ma ◽

Deli Wang ◽

Jian Weng ◽

Sheng Zhang ◽

Yuanshi Zhang

Keyword(s):

Interleukin 1 ◽

Interleukin 1 Beta ◽

Related Protein ◽

Interacting Protein ◽

Tnf Α ◽

Adenovirus E1b ◽

Factor Α ◽

Related Proteins ◽

Necrosis Factor ◽

Proinflammatory Factors

Abstract Background: Inflammation and apoptosis of chondrocytes are the pathological basis of osteoarthritis. Autophagy could alleviate the symptoms of inflammation and apoptosis. Previous study has shown that BCL2/adenovirus E1B 19 kDa protein-interacting protein 3 (BNIP3) can induce the occurrence and development of autophagy. However, it is unknown whether autophagy induced by BNIP3 can alleviate the inflammation and apoptosis of chondrocytes. Methods: We used the lentivirus to construct the overexpression BNIP3 chondrocytes. Next, the lipopolysaccharide (LPS) was used to stimulate these cells to simulate the physiological environment of osteoarthritis. After that, the enzyme-linked immunosorbent assays (ELISA) were performed to determine the levels of tumor necrosis factor-α (TNF-α), interleukin-1 beta (IL-1β) and interleukin-6 (IL-6) and the flow cytometry was performed to detect the apoptosis rates of chondrocytes. At last, the expression of autophagy related proteins was detected with the western blotting. Results: The expression of BNIP3 was suppressed after treatment with LPS. However, overexpression of BNIP3 inhibited the secretion of proinflammatory factors (TNF-α, IL-1β and IL-6) and decreased the apoptosis of chondrocytes. Furthermore, overexpression of BNIP3 led to the upregulation of autophagy related proteins expression including little computer 3 (LC3), autophagy-related protein 7 (ATG7) and Beclin-1. Application of autophagy inhibitor recovered the expression of proinflammatory factors and apoptosis rates of chondrocytes. Conclusions: BNIP3 decreased the LPS induced inflammation and apoptosis of chondrocytes by activating the autophagy.

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Regulation of Murine Protein C Gene Expression In Vivo: Effects of Tumor Necrosis Factor-α, Interleukin-1, and Transforming Growth Factor-β

Thrombosis and Haemostasis ◽

10.1055/s-0037-1614379 ◽

1999 ◽

Vol 82 (10) ◽

pp. 1297-1301 ◽

Cited By ~ 21

Author(s):

Takayoshi Shimokawa ◽

Tetsuhito Kojima ◽

David Loskutoff ◽

Hidehiko Saito ◽

Koji Yamamoto

Keyword(s):

Growth Factor ◽

Protein C ◽

Tumor Necrosis ◽

Tumor Necrosis Factor Α ◽

Transforming Growth Factor ◽

Interleukin 1 ◽

Transforming Growth Factor Β ◽

Tnf Α ◽

Factor Α ◽

Necrosis Factor

SummaryProtein C is a precursor of the anticoagulant serine protease, activated protein C, which inhibits coagulation factors Va and VIIIa. Although the liver appears to be the primary site of protein C synthesis, we previously demonstrated that the kidney and male reproductive organs also expressed abundant protein C mRNA in the mouse. In the present study, we further investigated the effects of tumor necrosis factor-α (TNF-α), interleukin-1 (IL-1), and transforming growth factor-β (TGF-β) on the expression of protein C mRNA in the principal producing organs, i.e., the liver, kidney, and testis. Both quantitative reverse transcription-PCR assay and in situ hybridization analysis revealed that TNF-α decreased protein C mRNA expression in the liver, kidney, and testis. IL-1 also down-regulated protein C mRNA expression in the liver and testis, but not in the kidney. In contrast, TGF-β unchanged the expression level of protein C mRNA in these three organs. These observations suggest that TNF-α and IL-1 may contribute to an increase in the procoagulant potential by down-regulation of protein C synthesis in the tissues during inflammatory processes.

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