Separation of Flavonoids and the Evaluation of the Anti-inflammatory Potential of Flavonoid-enriched Extracts from the Thlaspi arvense Linn by Inhibiting the Activation of TLR-4/NF-κ B

Abstract Background: Thlaspi arvense Linn, belonging to the dicotyledonous cruciferous family, is distributed Europe and Asia. In this study, we evaluated for the first time the anti-inflammatory effects of Thlaspi arvense Linn on LPS-stimulated RAW264.7 macrophages, and explored the related mechanism.Methods: The extract was identified and quantified using the HPLC, NMR. The anti-inflammatory activities of crude extracts C11, C12, C13 were screened by xylene-induced ear swelling and carrageenan-induced foot swelling in mice. The inflammatory mediators, pro-inflammatory cytokines and TLR-4-mediated signals in LPS-stimulated RAW264.7 macrophages were determined using NO activity assay, MTS, ELISA and Western blot.Results:The extract of Thlaspi arvense Linn was found to enrich flavonoid（mainly Orientin,Isoorientin,Vitexin;,Isovitexin,Luteolin-7-O-β-D-glucoside,Apigenin-7-O-β-D-glucoside,Luteolin,Apigeni）.5,7-dihydroxy-flavone-4′-(6′′-β-O-glucopyranoside)-β-O-D-glucopyranoside was novel, whereas isosapogenin and 8-methoxyvitexin were isolated for the first time from Thlaspi arvense Linn. The extract (flavonoid-enriched) inhibited xylene-induced ear swelling and carrageenan-induced foot swelling in mice. And suppressed LPS-induced overrelease of iNOS,TNF-α,COX-2 and IL-6 in RAW264.7 macrophages. The extract inhibited the inflammatory response through the signaling pathway mediated by TLR-4/NF- kappa B pathway and its downstream signals, IκB-α, NF-κB-P65 and IL-1β in LPS-stimulated macrophages.Conclusions: The present results demonstrate that the extract of Thlaspi arvense Linn inhibit inflammatory responses via the TLR-4/NF-KB-mediated signaling pathway.

Download Full-text

Punicalagin Prevents Inflammation in LPS-Induced RAW264.7 Macrophages by Inhibiting FoxO3a/Autophagy Signaling Pathway

Nutrients ◽

10.3390/nu11112794 ◽

2019 ◽

Vol 11 (11) ◽

pp. 2794 ◽

Cited By ~ 8

Author(s):

Cao ◽

Chen ◽

Ren ◽

Zhang ◽

Tan ◽

...

Keyword(s):

Western Blot ◽

Signaling Pathway ◽

Inflammatory Cytokines ◽

Blot Analysis ◽

Western Blot Analysis ◽

Inflammatory Responses ◽

Raw264.7 Macrophages ◽

Tnf Α ◽

Autophagy Inhibition ◽

Pro Inflammatory Cytokines

Punicalagin, a hydrolysable tannin of pomegranate juice, exhibits multiple biological effects, including inhibiting production of pro-inflammatory cytokines in macrophages. Autophagy, an intracellular self-digestion process, has been recently shown to regulate inflammatory responses. In this study, we investigated the anti-inflammatory potential of punicalagin in lipopolysaccharide (LPS) induced RAW264.7 macrophages and uncovered the underlying mechanisms. Punicalagin significantly attenuated, in a concentration-dependent manner, LPS-induced release of NO and decreased pro-inflammatory cytokines TNF-α and IL-6 release at the highest concentration. We found that punicalagin inhibited NF-κB and MAPK activation in LPS-induced RAW264.7 macrophages. Western blot analysis revealed that punicalagin pre-treatment enhanced LC3II, p62 expression, and decreased Beclin1 expression in LPS-induced macrophages. MDC assays were used to determine the autophagic process and the results worked in concert with Western blot analysis. In addition, our observations indicated that LPS-induced releases of NO, TNF-α, and IL-6 were attenuated by treatment with autophagy inhibitor chloroquine, suggesting that autophagy inhibition participated in anti-inflammatory effect. We also found that punicalagin downregulated FoxO3a expression, resulting in autophagy inhibition. Overall these results suggested that punicalagin played an important role in the attenuation of LPS-induced inflammatory responses in RAW264.7 macrophages and that the mechanisms involved downregulation of the FoxO3a/autophagy signaling pathway.

Download Full-text

Phytosterols Suppress Phagocytosis and Inhibit Inflammatory Mediators via ERK Pathway on LPS-Triggered Inflammatory Responses in RAW264.7 Macrophages and the Correlation with Their Structure

Foods ◽

10.3390/foods8110582 ◽

2019 ◽

Vol 8 (11) ◽

pp. 582 ◽

Cited By ~ 5

Author(s):

Yuan ◽

Zhang ◽

Shen ◽

Jia ◽

Xie

Keyword(s):

Nitric Oxide ◽

Inflammatory Responses ◽

Ester Group ◽

No Production ◽

Raw264.7 Macrophages ◽

Extracellular Signal ◽

Tnf Α ◽

Anti Inflammatory ◽

Factor Α ◽

Oxide Synthase

Phytosterols, found in many commonly consumed foods, exhibit a broad range of physiological activities including anti-inflammatory effects. In this study, the anti-inflammatory effects of ergosterol, β-sitosterol, stigmasterol, campesterol, and ergosterol acetate were investigated in lipopolysaccharide (LPS)-induced RAW264.7 macrophages. Results showed that all phytosterol compounds alleviated the inflammatory reaction in LPS-induced macrophage models; cell phagocytosis, nitric oxide (NO) production, release of tumor necrosis factor-α (TNF-α), and expression and activity of pro-inflammatory mediator cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), and phosphorylated extracellular signal-regulated protein kinase (p-ERK) were all inhibited. The anti-inflammatory activity of β-sitosterol was higher than stigmasterol and campesterol, which suggests that phytosterols without a double bond on C-22 and with ethyl on C-24 were more effective. However, inconsistent results were observed upon comparison of ergosterol and ergosterol acetate (hydroxy or ester group on C-3), which suggest that additional research is still needed to ascertain the contribution of structure to their anti-inflammatory effects.

Download Full-text

Rosanortriterpenes A–B, Two Promising Agents from Rosa laevigata var. leiocapus, Alleviate Inflammatory Responses and Liver Fibrosis in In Vitro Cell Models

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2020/8872945 ◽

2020 ◽

Vol 2020 ◽

pp. 1-9

Author(s):

Bai-Lin Li ◽

Juan-Juan Hu ◽

Jin-Dan Xie ◽

Chen Ni ◽

Hui-Jun Liang ◽

...

Keyword(s):

Liver Fibrosis ◽

Inflammatory Responses ◽

No Production ◽

Western Blot Assay ◽

Necrosis Factor Alpha ◽

Raw264.7 Macrophages ◽

Tnf Α ◽

Anti Inflammatory ◽

Rosa Laevigata

Rosanortriterpenes A–B (RTA and RTB), two nortriterpenoids, are characteristic constituents in the fruits of Rosa laevigata var. leiocapus. However, pharmacological studies on these compounds are still scarce. In the present study, we aim to investigate the anti-inflammatory mechanisms associated with the effects of RTA–B in RAW264.7 macrophages and LO2 cells by detecting cell viabilities, nitric oxide (NO) production, tumour necrosis factor-alpha (TNF-α), and interleukin-6 (IL-6) production. Simultaneously, the anti-inflammatory action mechanisms of these two compounds were illustrated through western blot assay. Besides, the antihepatic fibrosis activities of these compounds have also been explored. The results demonstrated that RTA and RTB inhibited the production of NO, TNF-α, and IL-6 and suppressed liver fibrosis. RTA and RTB treatment also greatly inhibited the activation of NF-kappaB (NF-κB) pathway. Our study confirmed the promising anti-inflammatory and anti-liver fibrosis actions of RTA–B, suggesting that they might be developed as alternative and promising drugs for the treatment of hepatic inflammatory and fibrotic diseases.

Download Full-text

Isoimperatorin exerts anti-inflammatory activity by targeting the LPS-TLR4/MD-2-NF-κB pathway

European Journal of Inflammation ◽

10.1177/20587392211000573 ◽

2021 ◽

Vol 19 ◽

pp. 205873922110005

Author(s):

Guirong Chen ◽

Yunong Liu ◽

Yubin Xu ◽

Mingbo Zhang ◽

Song Guo ◽

...

Keyword(s):

Molecular Docking ◽

Real Time ◽

Signaling Pathway ◽

Western Blotting ◽

Inflammatory Cytokines ◽

Real Time Pcr ◽

Inflammatory Responses ◽

Inflammatory Activity ◽

Tnf Α ◽

Anti Inflammatory

Isoimperatorin (QHS) is a phytoconstituent found in the methanolic extracts obtained from the roots of Angelica dahurica, which contains anti-inflammatory, anti-bacterial, analgesic, anti-tumor, and vasodilatory activities. QHS possesses potent antagonistic activity against lipopolysaccharide (LPS)-induced inflammation; however, the mechanism of action remains unclear. In this study, we investigated the anti-inflammatory effect of QHS and explored the underlying mechanisms. The QHS was purchased from Jiangsu Yongjian Pharmaceutical Co., Ltd. (Jiangsu, China). We performed MTT assay, real-time PCR, ELISA, and western blotting experiments to assess the anti-inflammatory activity and the possible mechanism of QHS in vitro. Molecular docking was performed to study the binding of QHS and myeloid differentiation protein-2 (MD-2) and elucidate the possible anti-inflammatory mechanism. QHS had no significant effect on cell viability. Moreover, pre-treatment with QHS significantly decreased the release of inflammatory cytokines and mediators including NO, TNF-α, IL-6, and IL-1β. In addition, real-time PCR showed that QHS decreased the mRNA expressions of iNOS, COX-2 TNF-α, IL-6, and IL-1β. Western blotting indicated that QHS could inhibit the expression of the proteins associated with the LPS-TLR4/MD-2-NF-κB signaling pathway. Lastly, molecular docking revealed a possible binding mechanism between QHS and MD-2. QHS exhibited anti-inflammatory activity when combined with MD-2, regulating the LPS-TLR4/MD-2-NF-κB signaling pathway, and inhibiting the release and expression of inflammatory cytokines and mediators. Furthermore, QHS can be used as a potential TLR4 antagonist, which blocks MD-2 binding, for treating inflammatory responses induced by LPS.

Download Full-text

Anemoside B4 protects against Klebsiella pneumoniae- and influenza virus FM1-induced pneumonia via the TLR4/Myd88 signaling pathway in mice

10.21203/rs.3.rs-27002/v2 ◽

2020 ◽

Author(s):

Jia He ◽

Renyikun Yuan ◽

Xiaolan Cui ◽

Yushun Cui ◽

Shan Han ◽

...

Keyword(s):

Influenza Virus ◽

Molecular Docking ◽

Signaling Pathway ◽

Therapeutic Effect ◽

Inflammatory Cytokines ◽

Inflammatory Activity ◽

Tnf Α ◽

Anti Inflammatory ◽

Myd88 Signaling ◽

Pro Inflammatory Cytokines

Abstract Background: Pneumonia refers to the inflammation of the terminal airway, alveoli and pulmonary interstitium, which can be caused by pathogenic microorganisms, physical and chemical factors, immune damage, and drugs. Anemoside B4, the major ingredient of Pulsatilla chinensis (Bunge) Regel, exhibited anti-inflammatory activity. However, the therapeutic effect of anemoside B4 on pneumonia has not been unraveled. This study aims to investigate that anemoside B4 attenuates the inflammatory responses in Klebsiella pneumonia (KP)- and influenza virus FM1 (FM1)-induced pneumonia mice model.Methods: The network pharmacology and molecular docking assays were employed to predict the targets of anemoside B4’s treatment of pneumonia. Two models (bacterial KP-infected mice and virus FM1-infected mice) were employed in our study. BALB/c mice were divided into six groups: control, model group (KP- induced pneumonia or FM1-induced pneumonia), anemoside B4 (B4)-treated group (2.5, 5, 10 mg/kg), and positive drug group (Ribavirin or Ceftriaxone Sodium Injection). Blood samples were collected for hematology analysis. The effects of B4 on inflammation-associated mediators were investigated by Enzyme-linked immunosorbent assay (ELISA) and hematoxylin and eosin staining (HE) staining. Proteins expression was quantified by western blotting.Results: The network results indicated that many pro-inflammatory cytokines such as tumor necrosis factor α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) participated in anemoside B4’s anti-inflammatory activity. The counts of neutrophil (NEU) and white blood cell (WBC), the level of myeloperoxidase (MPO), and the release of pro-inflammatory cytokines TNF-α, IL-1β, and IL-6 increased by KP or FM1 infection, which were reversed by anemoside B4. In addition, anemoside B4 significantly suppressed the FM1-induced expression of Toll-like receptor 4 (TLR4), myeloid differential protein-88 (MyD88), and myeloid differentiation protein-2 (MD-2), which were further validated by molecular docking data that anemoside B4 bound to bioactive sites of TLR4. Therefore, anemoside B4 exhibited a significant therapeutic effect on pneumonia via the TLR4/MyD88 pathway.Conclusion: Our findings demonstrated that anemoside B4 attenuates pneumonia via the TLR4/Myd88 signaling pathway, suggesting that anemoside B4 is a promising therapeutic candidate for bacterial-infected or viral-infected pneumonia.

Download Full-text

Triptolide exhibits anti-inflammatory effects on adipocytes and macrophages by inhibition of AMPK/mTOR pathway

10.21203/rs.3.rs-944266/v1 ◽

2021 ◽

Author(s):

Xue-li Li ◽

Zhao Liu

Keyword(s):

Signaling Pathway ◽

Therapeutic Potential ◽

Chinese Herb ◽

Mtor Signaling ◽

Low Grade ◽

Mtor Signaling Pathway ◽

Raw264.7 Macrophages ◽

Inflammatory Models ◽

Tnf Α ◽

Anti Inflammatory

Abstract Obesity is a growing global health problem and chronic over-nutrition disease with lipid accumulation that results in low-grade chronic inflammation in the microenvironment of adipose tissue. Triptolide is a diterpene lactone compound extracted from the roots of the Chinese herb TWHF and possesses a therapeutic potential due to its immunosuppressive and anti-inflammatory properties. In this study, we built obesity-related inflammatory models of adipocytes using LPS, Ma-CM and raw264.7 macrophages, while the obesity-related inflammatory models of macrophages were built using LPS and Ad-CM system. We used these inflammatory models to investigate the anti-inflammatory property of triptolide. Treatment of triptolide (0.005, 0.010, 0.020 and 0.040 μ M) inhibited LPS-induced or macrophages conditioned medium-stimulated activation of AMPK/mTOR signaling pathway (p < 0.05). The results showed that triptolide reduced the release of chemokines MCP-1, RANTES, EOTAXIN and KC in LPS, Ma-CM or RAW264.7 macrophages-stimulated 3T3-L1 adipocytes. Triptolide also diminished MCP-1, RANTES, EOTAXIN, KC and TNF-α in Ad-CM stimulated RAW264.7 macrophages, while expression of MCP-1, RANTES, TNF-α, GM-CSF and IL-6 was decreased in LPS stimulated RAW264.7 macrophages (p < 0.05). These results demonstrate that triptolide is not only effective against inflammatory response of RAW264.7 macrophages or 3T3-L1 adipocytes, but also disrupts the crosstalk between macrophages and adipocytes, particularly by inhibiting secretion of pro-inflammatory mediators through inhibiting the activation of AMPK/mTOR signaling pathway. Triptolide might benefit to ameliorate obesity-induced inflammatory diseases.

Download Full-text

Puerarin Prevents Acute Liver Injury via Inhibiting Inflammatory Responses and ZEB2 Expression

Frontiers in Pharmacology ◽

10.3389/fphar.2021.727916 ◽

2021 ◽

Vol 12 ◽

Author(s):

Junfa Yang ◽

Maomao Wu ◽

Hui Fang ◽

Yue Su ◽

Lingling Zhang ◽

...

Keyword(s):

Liver Injury ◽

Signaling Pathway ◽

Inflammatory Cytokines ◽

Inflammatory Responses ◽

Acute Liver Injury ◽

Expression Level ◽

Inflammatory Factors ◽

Tnf Α ◽

Zeb2 Expression ◽

Pro Inflammatory Cytokines

Puerarin, an isoflavone component extracted from herb radix puerariae, is widely used in China in the treatment of immune diseases and inflammation. Previous studies have demonstrated that puerarin prevented acute lung injury by regulating inflammatory responses. However, the effect of puerarin on acute liver injury (ALI) was unclear. The purpose of this study was to explore the beneficial effects of puerarin when applied to ALI. We found that puerarin inhibited liver injury and inflammatory cell infiltration in lipopolysaccharide (LPS)/D-galactose (D-Gal)-induced acute liver failure and the liver pro-inflammatory cytokines interleukin (IL)-1β, IL-6, and tumor necrosis factor-alpha (TNF-α) in liver tissues with ALI and LPS-induced L-02 cells but upregulated the expression level of zinc finger E-box-binding homeobox 2 (ZEB2). Significantly, the results of this study showed that the inhibition of liver pro-inflammatory cytokine (IL-1β, IL-6, and TNF-α) production in LPS-induced L-02 cells was caused by ZEB2 overexpression. However, knocking down ZEB2 promoted LPS-mediated secretion of liver pro-inflammatory cytokines in L-02 cells. Additional experiments showed that puerarin inhibited the activation of the NF-κB signaling pathway by elevating ZEB2 expression in L-02 cells. In summary, puerarin most likely prevented activation of the pro-inflammatory factors and reduced LPS/D-Gal-induced liver injury by enhancing the ZEB2 expression level and, consequently, blocking activation of the NF-κB signaling pathway in the liver.

Download Full-text

Anemoside B4 protects against Klebsiella pneumoniae- and influenza virus FM1-induced pneumonia via the TLR4/Myd88 signaling pathway in mice

10.21203/rs.3.rs-27002/v1 ◽

2020 ◽

Author(s):

Jia He ◽

Renyikun Yuan ◽

Xiaolan Cui ◽

Yushun Cui ◽

Shan Han ◽

...

Keyword(s):

Influenza Virus ◽

Molecular Docking ◽

Signaling Pathway ◽

Therapeutic Effect ◽

Inflammatory Cytokines ◽

Inflammatory Activity ◽

Tnf Α ◽

Anti Inflammatory ◽

Myd88 Signaling ◽

Pro Inflammatory Cytokines

Abstract Background Pneumonia refers to the inflammation of the terminal airway, alveoli and pulmonary interstitium, which can be caused by pathogenic microorganisms, physical and chemical factors, immune damage, and drugs. Anemoside B4, the major ingredient of Pulsatilla chinensis (Bunge) Regel, exhibited anti-inflammatory activity. However, the therapeutic effect of anemoside B4 on pneumonia has not been unraveled. This study aims to investigate that anemoside B4 attenuates the inflammatory responses in Klebsiella pneumonia (KP)- and influenza virus FM1 (FM1)-induced pneumonia mice model.Methods The network pharmacology and molecular docking assays were employed to predict the targets of anemoside B4’s treatment of pneumonia. Two models (bacterial KP-infected mice and virus FM1-infected mice) were employed in our study. BALB/c mice were divided into six groups: control, model group (KP- induced pneumonia or FM1-induced pneumonia), anemoside B4 (B4)-treated group (2.5, 5, 10 mg/kg), and positive drug group (Ribavirin or Ceftriaxone Sodium Injection). Blood samples were collected for hematology analysis. The effects of B4 on inflammation-associated mediators were investigated by Enzyme-linked immunosorbent assay (ELISA). Proteins expression was quantified by western blotting.Results The network results indicated that many pro-inflammatory cytokines such as tumor necrosis factor α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) participated in anemoside B4’s anti-inflammatory activity. The counts of neutrophil (NEU) and white blood cell (WBC), the level of myeloperoxidase (MPO), and the release of pro-inflammatory cytokines TNF-α, IL-1β, and IL-6 increased by KP or FM1 infection, which were reversed by anemoside B4. In addition, anemoside B4 significantly suppressed the FM1-induced expression of Toll-like receptor 4 (TLR4), myeloid differential protein-88 (MyD88), and myeloid differentiation protein-2 (MD-2), which were further validated by molecular docking data that anemoside B4 bound to bioactive sites of TLR4. Therefore, anemoside B4 exhibited a significant therapeutic effect on pneumonia via the TLR4/MyD88 pathway.Conclusion Our findings demonstrated that anemoside B4 attenuates pneumonia via the TLR4/Myd88 signaling pathway, suggesting that anemoside B4 is a promising therapeutic candidate for bacterial-infected or viral-infected pneumonia.

Download Full-text

20-Hydroxy-3-Oxolupan-28-Oic Acid Attenuates Inflammatory Responses by Regulating PI3K–Akt and MAPKs Signaling Pathways in LPS-Stimulated RAW264.7 Macrophages

Molecules ◽

10.3390/molecules24030386 ◽

2019 ◽

Vol 24 (3) ◽

pp. 386 ◽

Cited By ~ 5

Author(s):

Yufeng Cao ◽

Fu Li ◽

Yanyan Luo ◽

Liang Zhang ◽

Shuya Lu ◽

...

Keyword(s):

Gene Expression ◽

Molecular Mechanisms ◽

Nuclear Translocation ◽

Inflammatory Responses ◽

Nuclear Factor Κb ◽

Activator Protein 1 ◽

Raw264.7 Macrophages ◽

Tnf Α ◽

Mitogen Activated Protein ◽

Anti Inflammatory

20-Hydroxy-3-oxolupan-28-oic acid (HOA), a lupane-type triterpene, was obtained from the leaves of Mahonia bealei, which is described in the Chinese Pharmacopeia as a remedy for inflammation and related diseases. The anti-inflammatory mechanisms of HOA, however, have not yet been fully elucidated. Therefore, the objective of this study was to characterize the molecular mechanisms of HOA in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. HOA suppressed the release of nitric oxide (NO), pro-inflammatory cytokine tumor necrosis factor α (TNF-α), and interleukin 6 (IL-6) in LPS-stimulated RAW264.7 macrophages without affecting cell viability. Quantitative real-time reverse-transcription polymerase chain reaction (RT-qPCR) analysis indicated that HOA also suppressed the gene expression of inducible NO synthase (iNOS), TNF-α, and IL-6. Further analyses demonstrated that HOA inhibited the phosphorylation of upstream signaling molecules, including p85, PDK1, Akt, IκBα, ERK, and JNK, as well as the nuclear translocation of nuclear factor κB (NF-κB) p65. Interestingly, HOA had no effect on the LPS-induced nuclear translocation of activator protein 1 (AP-1). Taken together, these results suggest that HOA inhibits the production of cytokine by downregulating iNOS, TNF-α, and IL-6 gene expression via the downregulation of phosphatidylinositol 3-kinase (PI3K)/Akt and mitogen-activated protein kinases (MAPKs), and the inhibition of NF-κB activation. Our findings indicate that HOA could potentially be used as an anti-inflammatory agent for medical use.

Download Full-text

Piper sarmentosum extract alleviates inflammatory responses and improves barrier function in porcine intestinal epithelial (IPEC-J2) cells induced by lipopolysaccharide (LPS)

10.21203/rs.3.rs-17917/v1 ◽

2020 ◽

Author(s):

Dingfa Wang ◽

Luli Zhou ◽

zhou Hanlin ◽

Guanyu Hou

Keyword(s):

Protein Expression ◽

Signaling Pathway ◽

Inflammatory Responses ◽

Inflammatory Activity ◽

Complete Medium ◽

Metabolic Pathway Analysis ◽

Metabolic Pattern ◽

Tnf Α ◽

Anti Inflammatory ◽

Lps Treatment

Abstract Background: In this study, we investigated the anti-inflammatory effect of Piper sarmentosum extract (PSE) in the IPEC-J2 cells induced by lipopolysaccharide (LPS). Meanwhile, we also tested the metabolomics profile of cells treated by LPS and PSE. Method: IPEC-J2 cells (6×105 cell/well) were seeded on 6-well plates, and cells were divided into three treatments (control, LPS treatment and LPS + PSE-NB treatment). Each treatment was conducted in five replicates. After incubation for 24 h, cells in LPS + PSE-NB treatment were treated by media containing PSE-NB at 10 ug/ml (cells in control and LPS treatments were treated by complete medium). Cells were culture for 24 h, and cells in LPS treatment and LPS + PSE-NB treatment then were treated by media contain 1 μg/ml of LPS (cells in control was treated by complete medium) for another 24 h. After treatment, cells were used for gene expression assays, protein expression assays and metabolomics analysis.Results: We demonstrated that LPS stimulation significantly up-regulated the mRNA expression of IL-1, IL-6 and TNF-α (P < 0.05) compared with the control in the IPEC-J2 cells. Piper sarmentosum extract with n-butanol (PSE-NB) pre-treatment with 10 ug/mL before LPS stimulation significantly decreased the expression of IL-1, IL-6 and TNF-α compared with LPS treatment (P < 0.05). We found that PSE-NB improved the expression of intestinal tight junction proteins (ZO1 and Occludin) and NHE3 that were reduced by LPS stimulation (P < 0.05). Moreover, PSE-NB alleviated LPS-induced protein expression of p65 and p-p65 (P < 0.05) and inhibited the NF-κB signaling pathway. Metabolic pathway analysis indicated that PSE-NB exert anti-inflammatory activity mainly through affecting tryptophan metabolism. Its metabolic product, melatonin, has anti-inflammatory properties by inhibition of NF-κB activation, which consistent with our results regarding to anti-inflammatory activity of PSE-NB on inflammatory signaling pathway. Conclusion: These results suggested that PSE-NB might attenuate LPS-induced inflammatory responses in the IPEC-J2 cells by regulating inflammatory NF-κB signaling pathway and intracellular metabolic pattern.

Download Full-text