Expression Of Nuclear factor-κB Research Articles

Microrna-145 accelerates the inflammatory reaction through activation of NF-κB signaling in atherosclerosis cells and mice

Atherosclerosis (AS) is a chronic inflammation, which is a major cause of morbidity and mortality in the world. Accumulative evidences have demonstrated that miRNAs exert crucial roles in the development of AS. However, the effects of miR-145 and its underlying molecular mechanism remain incompletely clear. The aim of the present study is to explore the function of miR-145 in the occurrence and development of AS through investigating its role in inflammatory reactions. High-fat diet (HFD)-treated ApoE−/− mice were used as an in vivo model of atherosclerosis (AS). OxLDL-induced macrophages was employed as cell models of atherosclerosis. RT-PCR was used to evaluate the transfected efficiency of miR-145 mimic and inhibitor. RT-PCR and ELISA were performed to detect the expression of miR-145, and inflammatory cytokines such as tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), C-C motif chemokine ligand 2 (CCL-2), CCL-4 and CCL-7. Western blotting was used to evaluate the protein expression of nuclear factor κB (NF-κB) and its related proteins such as phosphorylated-signal transducer and activator of transcription 3 (p-STAT3), p-IκBα and acetylated p65 (ac-p65). Hematoxylin and eosin (H&E) staining were conducted to examine atherosclerotic lesion. Immunohistochemistry was carried out to detect the expression of α-smooth muscle Actin (α-SMA) and CD68. Luciferase reporter assay were carried out to examine the effect of miR-145 on the transcriptional activity of NF-κB. Our results showed that over-expression of miR-145 promoted the expression of IL-1β, TNF-α, CCL-2, CCL-4 and CCL-7 through promotion of NF-κB, p-IκBα, p-STAT3 and ac-p65 expression in vivo and in vitro. Besides, down-regulation of miR-145 expression relieved the aortic sinus lesion, increased the number of VSMCs and decreased the number of macrophages. In conclusion, our study demonstrated that miR-145 accelerated the inflammatory reaction through activation of NF-κB signaling in AS.

Rapamycin Alleviates Hormone Imbalance-Induced Chronic Nonbacterial Inflammation in Rat Prostate Through Activating Autophagy via the mTOR/ULK1/ATG13 Signaling Pathway.

Chronic prostatitis (CP) is a clinically common disease with high morbidity. It affects the patients' quality of life (QoL) as well as physical and mental health seriously due to the recurring symptoms of lower urinary tract and genitalia. As the opinions about the etiology of CP are still not uniform, it is very difficult to be treated or even cured. Autophagy is a highly conserved physiological function which is widely found in eukaryotic cells. In general, cells maintain a certain level of autophagy under physiological conditions, and the basal level of autophagy can be regulated by a variety of autophagy-related genes under stress such as hunger, infection, trauma, and other circumstances. Therefore, the main purpose of this study is to investigate the role of autophagy in chronic nonbacterial prostatitis (CNP, also called CP). In this paper, we established the CNP model via hypodermic injection of 17β-estradiol and subsequently abdominal rapamycin (a common autophagy inducer) treatment based on castrated rats. Then, the expression of nuclear factor-κB (NF-κB), interleukin-1β (IL-1β), and autophagy-related markers as well as autophagosome formation in prostate tissues, peripheral blood mononuclear cells (PBMCs), and serum of rats were evaluated respectively. In addition to some histological changes in the prostate tissues, we found the levels of NF-κB and IL-1β were significantly increased in the model group, along with significantly suppressed autophagy, whereas rapamycin could reverse these effects which involved in the mTOR/ULK1/ATG13 signaling pathway. In conclusion, our results suggested that rapamycin could ameliorate hormone imbalance-induced CNP by activating autophagy.

Tea Polyphenols Selectively Inhibit Proliferation and Migration of Airway Smooth Muscle Cells

Airway smooth muscle cells (ASMCs) have been recommended as a target for the treatment of inflammation and narrowing of the airways in asthma. New safe and efficient approaches to relieve symptoms caused by ASMCs is highly desired. Inspired by the inhibitory effects of tea polyphenols on vascular smooth muscle cells (VSMCs), tea polyphenols were applied in the current work to evaluate their regulation of ASMCs in vitro. A dose-dependent decrease of ASMCs density was observed after 24 h incubation with tea polyphenols. Additionally, ASMCs were significantly more sensitive to tea polyphenols than human bronchial epithelial cells (HBECs). Tea polyphenol treatment led to a dose dependent inhibition on ASMC migration and reduced the gene expression of nuclear factor-κB (NF-κB). In studies which compared the four main polyphenolic constituents of tea polyphenols—including epicatechin (EC), epicatechingallate (ECG), epigallocatechin (EGC) and epigallocatechingallate (EGCG)—on the proliferation of ASMCs, EGC was identified as being the most potent. These results suggest tea polyphenols are a promising agent for ASMCs targeted asthma control.

The expression of Nuclear Factor-κB in a rat model of polycystic ovary syndrome

Objective: To discuss the effect of Nuclear Factor-κB (NF-κB) in polycystic ovary syndrome (PCOS). Materials and Methods: Thirty female SD rats were divided into a control group (n=15) and a PCOS group (n=15). The expression of NF-κB in ovaries of each group was monitored by way of immunohistochemistry and quantitative PCR. Results: The expression of NF-κB p65 in ovaries by immunohistochemical and fluorescence quantitative PCR shows no significant difference between the two groups (p > 0.01). Conclusion: The pathogenesis of this research model has no obvious association with NF-κB.

Effect of transient ischemic attack on early neurological deterioration in patients with ipsilateral ischemic stroke

Objective: To explore the effect of anterior circulation transient ischemic attack (TIA) on early neurological deterioration (END) in patients with ipsilateral ischemic stroke and its mechanism. Methods: (1) One hundred and thirteen patients with ipsilateral ischemic stroke and 36 healthy volunteers (healthy control group) in Neurology Department of Tianjin Medical University General Hospital from November 2014 to July 2015 were recruited into this study. According to whether got TIA before ischemic stroke, patients were divided into simple ischemic stroke group (CI group, n=87) and TIA-CI group (n=26). Their END, NIHSS score, 3-month mRS score, infarct size, serum hs-CRP and other risk factors were compared. (2) The peripheral blood mononuclear cells (PBMCs) were extracted from peripheral blood of TIA-CI group and CI group at 24 h, the 3 th day, the 7 th day and 12th day after ischemic stroke onset. At the same time, PBMCs of control group were collected. Western blot was carried out to evaluate the expression of nuclear factor-κB (NF-κB). Immunofluorescence was used to detect the cytoplasmic-to-nuclear shuttling of NF-κB. Results: (1) The incidence of END, NIHSS score at discharge, 3-month mRS score and serum hs-CRP level were significantly lower whereas the infarct size was significantly smaller of TIA-CI group than CI group (11.5% vs 31.0%, 1.9±2.3 vs 3.3±3.7, 0.9±0.8 vs 1.8±1.8, 1.1(0.3, 2.5) cm(3) vs 2.4(0.5, 22.8) cm(3,) (2.5±3.2) mg/L vs (6.2±3.2) mg/L, all P<0.05) . hs-CRP was positively correlated with END (r=0.311, P<0.05). (2) Expression of NF-κB: ① Compared with control group, the NF-κB expression increased first and then decreased in both of the two patient groups, and it decreased earlier in TIA-CI group than CI group.②In each time point, NF-κB expression of TIA-CI group was lower than CI group(t=1.754, P<0.05; t=1.858, P<0.05; t=0.609, P<0.05; t=0.519, P<0.05). (3) Activity of NF-κB: Most of NF-κB were inactivation and located in cytoplasm of control group. NF-κB of TIA-CI group and CI group was activated and translocated from cytoplasm into nuclear at the 24 h and 3 th day after ischemic stroke. At the 7 th day and 12th day, the accumulation of NF-κB in nuclear decreased and most of them located in cytoplasm in TIA-CI group, whereas most of NF-κB still located in nuclear in CI group. The activated station of NF-κB lasted shorter in TIA-CI group than CI group. Conclusions: TIA can reduce the incidence of END in patients of ipsilateral ischemic stroke. Its protective effect may relate with the inhibition of inflammation which induced by NF-κB.

Unfractionated heparin inhibits the activation of endothelial cells by decreasing the degree of HOXA9 reduced expression

To study unfractionated heparin (UFH) effect on the expression of HOXA9 in activation of human umbilical vein endothelial cells (HUVECs) induced by lipopolysaccharide (LPS). HUVECs were cultured and they were randomly divided into four groups (n = 5) for the challenge respectively: (1) control group (with an equal volume of phosphate buffer saline); (2) LPS group (LPS 10 mg/L); (3) UFH group (UFH 10 kU/L); (4) UFH+LPS group (10 kU/L UFH 30 minutes + LPS 10 mg/L). After treatment for 3 hours, the expressions of HOXA9, E-selectin and nuclear factor-κB (NK-κB) in endothelial cells were detected by Western Blot. Compared with the control group, the expression of HOXA9 in LPS group was significantly decreased, the expressions of E-selectin and NF-κB were significantly increased (HOXA9/β-actin: 0.082±0.009 vs. 0.199±0.067, E-selectin/β-actin: 0.113±0.055 vs. 0.047±0.030, NF-κB/β-actin: 0.845±0.025 vs. 0.664±0.092, all P < 0.05). Compared with LPS group, the expression of HOXA9 in UFH+LPS group was significantly increased, the expressions of E-selectin and NF-κB were significantly decreased (HOXA9/β-actin: 0.190±0.096 vs. 0.082±0.009, E-selecin/β-actin: 0.057±0.017 vs. 0.113±0.055, NF-κB/β-actin: 0.544±0.060 vs. 0.845±0.025, all P < 0.05). Each protein expression of UFH group were in accordance with the control group. In LPS stimulated endothelial cells, HOXA9 expression is down regulated, E-expression is reduced, and endothelial cell activation is inhibited. UFH can inhibit the activation of endothelial cells by decreasing the degree of HOXA9 reduced expression.

Effect of mild hypothermia on expression of inflammatory factors in surrounding tissue after minimally invasive hematoma evacuation in the treatment of hypertensive intracerebral hemorrhage.

Mild hypothermia combined with minimally invasive hematoma evacuation was evaluated in the treatment of hypertensive intracerebral hemorrhage to reduce inflammatory response of brain tissue around hematoma and ameliorate brain function, and to investigate its safety, effectiveness and feasibility. A total of 206 patients with acute spontaneous hypertensive intracerebral hemorrhage were collected clinically and randomly divided into minimally invasive hematoma evacuation group (group A) and mild hypothermia combined with minimally invasive hematoma evacuation (group B). The National Institutes of Health Stroke Scale (NIHSS) score was used before and after treatment. Group A was treated with minimally invasive intracranial hematoma evacuation using intracranial hematoma grinding puncture needle while group B received whole body water circulation type cooling blanket plus local cerebral mild hypothermia therapy with ice cap on the basis of minimally invasive surgery. Patients brain tissue fragments around hematoma taken out with rinsing during operation and at postoperative 1, 3 and 7 days were investigated. The contents of tumor necrosis factor-α (TNF-α) in serum at postoperative 1, 3 and 7 days were evaluated by enzyme-linked immunosorbent assay (ELISA). For the degree of nerve function defect of patients in the two groups, NIHSS score was lower in group B than that in group A at days 3 and 7, and the differences were statistically significant (P<0.05). The serum TNF-α content and expression of nuclear factor-κB (NF-κB) in brain tissue around hematoma reached the peak on the 3rd day. The TNF-α content and NF-κB expression were lower in group B than those in group A at each time-point (P<0.05). Mild hypothermia combined with minimally invasive hematoma evacuation can reduce the damage of hematoma to the surrounding brain tissue, effectively alleviate inflammatory response and decrease brain tissue injury, thus ameliorating brain function.

Downregulation of Frizzled-7 induces the apoptosis of hepatocellular carcinoma cells through inhibition of NF-κB.

The aim of the present study was to investigate the functional role of Frizzled-7 (FZD7) in the apoptosis of hepatoma cells. HepG2 and Huh-7 hepatocellular carcinoma (HCC) cell lines with FZD7 expression were selected for use in the present study. The small hairpin RNA (shRNA) eukaryotic expression vector specific to FZD7 was constructed using gene recombination, and was then transfected into HepG2 and Huh-7 hepatoma cell lines using Lipofectamine 2000 to assess whether the downregulation of FZD7 could affect the proliferative ability of these cells. The results demonstrated that the downregulation of FZD7 expression significantly inhibited the proliferative ability of both cell types through the induction of cell apoptosis, as evidenced using Cell Counting kit-8 assays and flow cytometry. Furthermore, the western blotting results demonstrated that silencing of FZD7 increased the activities of caspase-3 and caspase-9. These increases were also associated with the downregulation of the inhibitor of the apoptosis protein family. Additionally, it was revealed that silencing of FZD7 expression caused the downregulation of apoptosis regulator Bcl-2 and Bcl-XL in HepG2, and Huh-7 cells, as determined through western blot analysis and reverse transcription-quantitative polymerase chain reaction. In the following work, ELISA and western blot analysis revealed that the knockdown of FZD7 inhibited the expression and activities of nuclear factor-κB (NF-κB) p65. Furthermore, it was demonstrated that the expression levels of phosphylated-Smad2/3 were markedly upregulated in sh-FZD7-transfected HepG2 and Huh-7 cells. Then, shRNA eukaryotic expression vector specific to transforming growth factor (TGF)-β receptor II was transfected into both cell lines to investigate the association between the TGF-β/Smad signaling pathway and NF-κB p65. Notably, when the TGF-β/Smad signaling pathway was inhibited, no significant differences in the cell apoptosis rate and NF-κB expression levels were identified in HCC cells. Overall, the results of the present study suggest that the shRNA-mediated knockdown of FZD7 induces apoptosis of hepatoma cell lines through the inhibition of NF-κB. In addition, the TGF-β/Smad signaling pathway appeared to partially participate in the underlying molecular mechanism of FZD7 in HCC.

Nano-gold displayed anti-inflammatory property via NF-kB pathways by suppressing COX-2 activity

Rheumatoid arthritis (RA) is an autoimmune inflammatory disease, affecting almost 1% of world population. Although the exact cause of RA is not known but the complex interaction between inflammatory mediators like tumour necrosis factor-α (TNF-α), interleukin-1β (IL-1β), cyclooxygenase-2 (COX-2) and nitric oxide (NO) is accountable for cartilage destruction in joints. Gold is used for arthritis treatment since long without knowing its mechanism of action. Hence, the present study was designed to assess antiarthritic activity of nanogold (AuNGs) in collagen-induced arthritic (CIA) rat model by virtue of decreasing inflammatory mediators and oxidative stress. After induction CIA rats were treated with AuNGs in phosphate buffer at a dose of 20 μg/kg body weight for 20 days and found a significant decrease in the level of inflammatory mediators like TNF-α, IL-1β, COX-2 and transcription factor NF-kB (Nuclear factor-kB), which was found to be elevated in CIA rats. Additionally imbalance in oxidant and antioxidant status were determined and perceived that AuNGs remarkably attenuates the imbalance in level of antioxidant and oxidant near to normal. In consistent to biochemical results, mRNA expression of NF-kB, TNF-α, COX-2, and iNOS were also up-regulated in CIA rats, which were considerably down regulated by AuNGs treatment. These findings were positively correlated with the histological results of joints, displayed reduced inflammation and bone erosion in treated group. This study demonstrates the ability of AuNGs to ameliorate production of inflammatory mediators and oxidative stress in CIA rats. Induction of arthritis in rats showed increased inflammation, which activate the transcription factor NF-kB through activation of of IkB kinases (IKK) and ubiquination/proteosome degradation of IKB and transportation of activated NF-kB from cytoplasm to nucleus. In nucleus activated NF-kB bind to the promoter region of target gene and up regulate the production of pro-inflammatory cytokines, COX-2 and other inflammatory mediators that leads to cartilage destruction. AuNGs inhibit the activation of NF-kB and other inflammatory mediators and attenuate inflammation and cartilage destruction. COX-2: cyclooxygenase-2; IKK: IkB kinases; IKB: I Kappa B; IL-1β: interleukin-6; IL-6: interleukin-6; iNOS: inducible nitric oxide synthase; NF-kB: nuclear transcription factor kappa B; ROS: reactive oxygen species; TNF-α: tumour necrosis factor-alpha.

Berberine hydrochloride inhibits cell proliferation and promotes apoptosis of non-small cell lung cancer via the suppression of the MMP2 and Bcl-2/Bax signaling pathways.

Berberine, also known as berberine hydrochloride and isoquinoline alkaloid, is a major alkaloid from Coptis chinensis. Berberine's extensive biological properties have previously been studied, and it has been used clinically for the treatment of diarrhea, hypertension, diabetes and other diseases. The present study aimed to determine the possible anticancer effects of berberine hydrochloride treatment on human non-small cell lung cancer (NSCLC) cell proliferation and apoptosis via the matrix metalloproteinase 2 (MMP-2) and the B-cell lymphoma 2 (Bcl-2)/Bcl-2-associated X protein (Bax) signaling pathway. Human A549 lung carcinoma cells were exposed to various concentrations of berberine hydrochloride in order to analyze the possible anticancer effects on NSCLC cell proliferation and apoptosis, using a MTT assay and an Annexin V-fluorescein isothiocyanate/propidium iodide apoptosis kit. Subsequently, the present study detected the expression of MMP-2, Bcl-2, Bax and Janus kinase 2 (Jak2). Berberine hydrochloride treatment inhibited the expression of vascular endothelial growth factor (VEGF) and nuclear factor κB (NF-κB) and transcription factor AP-1 (AP-1) proteins, in A549 cells. Firstly, it was revealed that berberine hydrochloride treatment may inhibit proliferation, increase cytotoxicity and enhance apoptosis in A549 cells. Subsequently, treatment with berberine hydrochloride significantly downregulated MMP-2 protein expression, increased the activity of the Bcl-2/Bax signaling pathway and suppressed the Jak2/VEGF/NF-κB/AP-1signaling pathways. These results suggest that berberine hydrochloride may be a potential novel anticancer drug, since it inhibits cell proliferation and promotes the rate of apoptosis of NSCLC cells by the suppression of the MMP-2, Bcl-2/Bax and Jak2/VEGF/NF-κB/AP-1 signaling pathways.

Juglanin suppresses fibrosis and inflammation response caused by LPS in acute lung injury

Acute lung injury in children is a complication showing devastating disorders linked to fibrosis progression and inflammation response. Fibrosis and inflammation response are two markers for acute lung injury. Juglanin is a natural product mainly isolated from green walnut husks of Juglans mandshurica, which isconsidered as the functional composition among a series of compounds. It exhibited effective role in various diseases by inhibiting inflammation response. In our study, the protective effects and anti-inflammatory activity of juglanin were investigated in mice and lung cells treated by lipopolysaccharide (LPS) to reveal the possible mechanism by which juglanin attenuates acute lung injury. The mice were separated into four groups. The mouse model was established with 15 mg/kg LPS injection. Juglanin dramatically reduced the inflammation of cell infiltration. Compared to mice only treated with LPS, LPS-treated mice in the presence of juglanin developed less lung fibrosis with lower levels of α-smooth muscle-actin (α-SMA), collagen type I, collagen type III, and transforming growth factor-β1 (TGF-β1). Additionally, juglanin markedly downregulated inflammatory cytokine secretion and phosphorylated nuclear factor-κB (NF-κB) expression via inhibiting IKKα/IκBα signaling pathway. Our results indicate that juglanin has a protective role in LPS-triggered acute lung injury via suppression of fibrosis and inflammation response by NF-κB signaling pathways inactivation. Thus, juglanin may be a potential candidate as dietary supplement for acute lung injury for children in future.

Effects of dexmedetomidine on inflammatory factors, T lymphocyte subsets and expression of NF-κB in peripheral blood mononuclear cells in patients receiving radical surgery of colon carcinoma.

The effects of dexmedetomidine on inflammatory factors, T lymphocyte subsets and expression of nuclear factor-κB (NF-κB) in peripheral blood mononuclear cells in patients receiving radical surgery of colon carcinoma were investigated. A total of 141 patients receiving radical surgery of colon carcinoma from January 2014 to April 2017 were divided into two groups randomly. The patients in the treatment group were given dexmedetomidine, while the patients in the control group were treated with saline. Results showed that there were no significant differences in preoperative levels of NF-κB, sICAM-1 and IL-8 between the two groups. However, the above three indexes of the groups were all significantly increased at 0.5 and 24 h after operation, and the results showed that the control group had a higher degree of increase (P<0.05). It was also found that the changes in levels of IL-6 and CRP in patients were the same as those of the above three indexes; in other words, the degree of the increase in the control group was significantly higher than that in the treatment group after operation. Moreover, it was found that there was little difference in the preoperative Ramsay score of patients between the two groups. The scores at 0.5 and 12 h after operation in the treatment group were significantly higher than those in the control group (P<0.05). It was also found that the intraoperative and postoperative dosages of fentanyl in the treatment group were significantly less than those in the control group (P<0.05). We can conclude that the application of dexmedetomidine during anesthesia in patients receiving radical operation of colon carcinoma has a better clinical treatment effect, which can reduce the secretion of inflammatory factors, decrease the inhibition of immunity and reduce the use of fentanyl.

The effect of 1,25 dihydroxyvitamin D3 on HCl/Ethanol-induced gastric injury in rats.

This study evaluated ulceroprotective and antioxidant effect of 1,25 dihydroxyvitamin D3 against gastric damage in rats. Rats were treated intraperitoneally with either 1,25 dihydroxyvitamin D3 (0.25 μg/kg) or saline for 14 days. On day-15, the non-selective cyclooxygenase inhibitor indomethacin (10 mg/kg; subcutaneously), the inhibitor of sulfhydryl groups N-ethylmaleimide (10 mg/kg; intraperitoneally) or ATP-sensitive K+ channel blocker glibenclamide (10 mg/kg; orally) was given prior to 1,25 dihydroxyvitamin D3. Animals were euthanized at 60 min post ulcerogenic challenge (0.3 M HCl and 60% ethanol (0.2 mL; orally). Stomach and blood were collected for biochemical and histological evaluations. HCl/Ethanol group revealed severely damaged mucous and glandular epithelium with diffuse hemorrhage and inflammatory cell infiltration (microscopic score: 10.67 ± 0.67 and ulcer index: 33.13 ± 5.09). 1,25 dihydroxyvitamin D3 decreased the extent of damage (microscopic score: 6.80 ± 0.02 and ulcer index: 19.00 ± 4.34; p < 0.05), and the elevations in gastric malondialdehyde level (p < 0.001), myeloperoxidase activity (p < 0.001), nuclear factor-κB expression (p < 0.05), and apoptotic index (p < 0.05) following HCl/Ethanol challenge. Decreased gastric glutathione following HCl/Ethanol administration was restored by 1,25 dihydroxyvitamin D3 (p < 0.01). These findings demonstrated protection of the gastric mucosa against HCl/Ethanol-induced injury by 1,25 dihydroxyvitamin D3 via attenuation of inflammatory reaction, oxidative stress and apoptosis.

Deciphering PPARγ activation in cardiometabolic syndrome: studies by in silico and in vivo experimental assessment

Cardiometabolic syndrome (CMetS) is a consolidation of metabolic disorders characterized by insulin resistance, dyslipidemia and hypertension. Curcumin, a natural bioactive compound, has been shown to possess notable anti-oxidant activity and it has also been included as a super natural herb in the super natural herbs database. Most of the beneficial effects of Curcumin are possibly due to activation of the nuclear receptor, peroxisome proliferator-activated receptor gamma (PPARγ). The present study investigates molecular interactions of curcumin with PPARγ protein through molecular docking and molecular dynamics (MD) simulation studies. Further, effect of curcumin on high fat diet induced CMetS was studied in rats along with western blot for PPARγ and nuclear factor-κB (NF-κB) expressions and histopathological studies. Computational studies presented several significant molecular interactions of curcumin including Ser289, His323, His449 and Tyr473 of PPARγ. The in vivo results further confirmed that curcumin was able to ameliorate the abnormal changes and also, increased PPARγ expressions. The results confirm our hypothesis that activation of PPARγ by curcumin possesses the therapeutic potential to ameliorate the altered levels of metabolic changes in rats in the treatment of CMetS. This is the first report of CMetS treatment by curcumin and study of its underlying mechanism through in silico as well as in vivo experiments.

Andrographolide derivative CX-10 ameliorates dextran sulphate sodium-induced ulcerative colitis in mice: Involvement of NF-κB and MAPK signalling pathways

Ulcerative colitis (UC) is a type of inflammatory bowel disease (IBD), which is characterized by chronic intestinal inflammation and leads to an increased risk of colon cancer. There are many studies using phyto-ingredients as a novel approach for the treatment of UC. The plant Andrographis paniculata (Acanthaceae) is a safe and edible vegetable that has been extensively adopted in traditional Chinese medicine for conditions involving inflammation, and the most active phytochemical agent is andrographolide. The andrographolide derivative 3,14,19-triacetyl andrographolide, which is known as CX-10 (a hemi chemical synthesized from andrographolide), has been found to possess strong anti-inflammatory properties. In the present study, we investigated the therapeutic potential of CX-10 as a complementary and alternative medicine against dextran sulphate sodium (DSS)-induced ulcerative colitis in mice. Our results revealed that CX-10 treatment reduced body weight loss, reduced colon length shortening, decreased colon weight, decreased the spleen index, decreased the disease activity index (DAI), and alleviated histological damage in the colon. The expression of TNF-α and IL-6 and the activity of myeloperoxidase (MPO) in colonic tissues were significantly reduced in CX-10 supplemented mice. It is noteworthy that the efficacy of 200 mg/kg of CX-10 was equivalent to that of the mesalazine positive control (200 mg/kg). Furthermore, western blot analysis revealed that CX-10 treatment reduced the expression of nuclear factor-κB (NF-κB) p65 and p-IκBα, increased the expression of IκBα and down-regulated the phosphorylation of p38 mitogen-activated protein kinase (MAPK), ERK and JNK. In conclusion, CX-10 treatment attenuated DSS-induced UC in mice through inhibiting the activation of NF-κB and MAPK pathways and reducing TNF-α and IL-6 levels, suggesting that CX-10 is a potential therapeutic drug for UC.

Antiproteinuric effect of an endothelin-1 receptor antagonist in puromycin aminonucleoside-induced nephrosis in rat.

BackgroundThe pathogenesis of idiopathic nephrotic syndrome (INS) remains unclear, although recent studies suggest endothelin 1 (ET-1) and CD80 of podocytes are involved. We investigated the potential of antagonist to ET-1 receptor type A (ETRA) as therapeutic agent through the suppression of CD80 in a rat model of INS.MethodsPuromycin aminonucleoside (PAN) was injected to Wister rats to induce proteinuria: some were treated with ETRA antagonist and others were treated with 0.5% methylcellulose. Blood and tissue samples were collected. Quantitative PCR was used to determine the expression of Toll-like receptor-3 (TLR-3), nuclear factor-κB (NF-κB), CD80, talin, ETRA, and ET-1 in the kidney. To confirm the level of CD80 protein expression, immunofluorescence staining and western blot analysis of the renal tissue were performed.ResultsAmount of proteinuria in the treatment group was significantly lower than the other groups. The same-day body weight, serum creatinine values, and blood pressure were not significantly different. ETRA antagonist restores podocyte foot process effacement as well as the aberrant expression of TLR-3, nuclear factor-κB (NF-κB), and CD80 in PAN-injured kidneys.ConclusionsThe ETRA antagonist may be promising drug for INS as it showed an antiproteinuric effect. Its action was considered to be through suppression of CD80 expression on podocytes.

Protective effect and molecular mechanism of liquiritin on oxybuprocaine-induced apoptosis of human corneal endothelial cells.

This study was designed to investigate the protective effect and possible molecular mechanism of liquiritin on oxybuprocaine-induced apoptosis of human corneal endothelial cells (HCECs). In this study, the effect of oxybuprocaine on the proliferation of HCEC-12 was detected using cell counting kit-8 (CCK-8). The inductive effect of oxybuprocaine on HCEC-12 apoptosis and protective effect of liquiritin against oxybuprocaine-induced HCEC-12 apoptosis were tested by Annexin V/propidium iodide (PI) staining and flow cytometry. The production of reactive oxygen species (ROS) was analyzed by 2,7-dichlorodi-hydrofluorescein diacetate (DCFH-DA) staining and fluorescent-activated cell sorting (FACS), and the expression of nuclear factor-κB (NF-κB) p65 and apoptosis-related proteins, caspase-3 and Bax, was determined by western blot analysis. Our results show that liquiritin resisted the inhibitory effect of oxybuprocaine on the proliferation of HCEC-12, and cell activity had the most significant increase in pretreatment with liquiritin group in the concentration of 8 mg/ml; compared with that in oxybuprocaine group. Apoptosis in pretreatment with liquiritin was distinctly decreased and liquiritin resisted the production of ROS in HCEC-12 induced by oxybuprocaine. Investigation of molecular mechanism revealed that the pretreatment with liquiritin and pyrrolidinedithiocarbamic acid (PDTC) obviously blocked the expression of NF-κB p65 in nuclear protein increased by oxybuprocaine and the expression levels of total proteins, caspase-3 and Bax.Moreover, tumor necrosis factor-α (TNF-α) blocked the inhibitory effect of liquiritin on the expression of NF-κB p65 in nuclear protein and total proteins, caspase-3 and Bax, thus obstructing the protective effect of liquiritin on corneal epithelial cells. The results of this study indicated that liquiritin reduces the expression of apoptosis protein and increases the expression of anti-apoptotic protein through inhibiting NF-κB signal pathway, thus resisting HCEC-12 apoptosis induced by oxybuprocaine.

Thymoquinone alleviates the experimentally induced Alzheimer's disease inflammation by modulation of TLRs signaling.

Alzheimer's disease (AD) is characterized by a robust inflammatory response elicited by the accumulation and deposition of amyloid-β (Aβ) within the brain. Aβ induces detrimental inflammatory responses through toll-like receptors (TLRs) signaling pathway. Thymoquinone (TQ), the main active constituent of Nigella sativa oil, has been reported by several previous studies for its potent anti-inflammatory effect. The aim of this study is to elucidate the effect of TQ in improving learning and memory, using a rat model of AD induced by a combination of aluminum chloride (AlCl3) and d-galactose (d-Gal). TQ was administered orally at doses of 10, 20, and 40 mg/kg/day for 14 days after AD induction. Memory functions were assessed using the step through passive avoidance test. Amyloid plaques were shown to be present using hematoxylin and eosin staining. Tumor necrosis factor-alpha (TNF-α) and Interleukin-1beta (IL-1β) levels in brain were assessed via ELISA and profiling TLR-2, TLR-4, myeloid differential factor 88, toll-interleukin-1 receptor domain-containing adapter-inducing interferon-β, interferon regulatory factor 3 (IRF-3), and nuclear factor-κB (NF-κB) expressions via real-time polymerase chain reaction. TQ improved AD rat cognitive decline, decreased Aβ formation and accumulation, significantly decreased TNF-α and IL-1β at all levels of doses and significantly downregulated the expression of TLRs pathway components as well as their downstream effectors NF-κB and IRF-3 mRNAs at all levels of doses ( p < 0.05). We concluded that TQ reduced the inflammation induced by d-Gal/AlCl3 combination. It is therefore reasonable to assign the anti-inflammatory responses to the modulation of TLRs pathway.

Amelioration of inflammatory responses by Socheongryong-Tang, a traditional herbal medicine, in RAW 264.7cells and rats.

Socheongryong-Tang (SCRT) is a natural medicine prescription that has been mainly used in East Asia for the treatment of inflammatory disorders, including asthma and allergic rhinitis. The present study evaluated the anti-inflammatory effects of SCRT on lipopolysaccharide (LPS)-stimulated RAW 264.7 cells and in a rat model of carrageenan (CA)-induced paw edema. Levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6 and prostaglandin E2 (PGE2) in the culture supernatant were quantified and nitric oxide (NO) production was monitored. In addition, the effect of SCRT on the protein expression of nuclear factor-κB (NF-κB), mitogen-activated protein kinases (MAPKs), inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) was assessed by western blot analysis. Furthermore, the effects of SCRT on acute inflammation in vivo and changes in the histomorphometry and histopathology of paw skin were observed using CA-treated rats. SCRT (1 mg/ml) inhibited the LPS-induced changes in the protein expression of NF-κB, JNK, ERK1/2, iNOS and COX-2, as well as the production of NO, PGE2 and cytokines. In the rat paw edema assay, administration of 1 g/kg of lyophilized powder obtained from the aqueous extracts of SCRT for 3 consecutive days inhibited the CA-induced increases in skin thickness, mast cell degranulation, and infiltration of inflammatory cells in the ventral and dorsal pedis skin within 4 h. These results demonstrated that SCRT exerts its anti-inflammatory activities in LPS-stimulated RAW 264.7 cells through decreasing the production of inflammatory mediators, including PGE2, NO and cytokines, via suppression of the NF-κB and JNK and ERK1/2 signaling pathways. In addition, the data of the CA-induced paw edema indicated an anti-edema effect of SCRT. SCRT (1 g/kg) reduced acute edematous inflammation through inhibition of mast cell degranulation and infiltration of inflammatory cells. Therefore, the present study provided scientific evidence for the anti-inflammatory activities of SCRT as well as the underlying mechanisms.

Protective effects of amifostine, curcumin and caffeic acid phenethyl ester against cisplatin-induced testis tissue damage in rats.

Cisplatin is an effective antineoplastic drug that is usually used to treat a number of different types of cancer in the clinic. One of the most notable side effects of cisplatin use is infertility. The present study was designed to determine the non-oxidative testicular effects caused by the use of cisplatin in rats. The rats were randomly allocated to the experimental groups. The untreated rats represented the control group (group I) and the treatment groups were as follows: cisplatin alone (group II), cisplatin+amifostine (group III), cisplatin+curcumin (group IV), and cisplatin+caffeic acid phenethyl ester (CAPE; group V). The present study observed that following cisplatin administration, the expression of nuclear factor-κB (NF-κβ)/p65, caspase-3 and 8-deoxyguanosine (8-OHdG) increased in germinal epithelium and Leydig cells. However, the expression of these markers decreased in groups III–V, most notably in the group treated with amifostine. cisplatin induced-damage was countered by amifostine and curcumin. The results revealed that the activation of NF-κB, caspase-3 and 8-OHdG had a significant role in cisplatin-induced testicular toxicity. Thus, amifostine, curcumin and, to a lesser extent, CAPE have the potential for use as therapeutic adjuvants in cisplatin-induced testis injury.