Tsui-Ting Ching et al. (2007) International Worm Meeting "Celecoxib Extends Worm Lifespan Independent of COX-2 Inhibition."

Tsui-Ting Ching, Ching-Shih Chen, Ao-Lin Hsu, Wei-Chung Chiang

[International Worm Meeting, 2007]

The goal of our research is to identify small molecules that can delay aging and prolong the healthy period of human lifespan. As the first step toward this goal, we carried out a pilot screen for small molecules that can extend the lifespan of C. elegans. From the screen, we found that exposure of worms to a low dosage of celecoxib can significantly extend the animals lifespan by up to 16%. We also found that exposure to celecoxib just during the adulthood is sufficient to extend lifespan. Celecoxib is known as a non-steroidal anti-inflammatory drug (NSAID), whose anti-inflammatory activity is achieved by inhibiting the activity of cyclooxygenases (COXs). It has also been shown that long-term use of celecoxib can reduce the risk of developing cancer by inducing apoptosis in cancer cells. Recent studies have suggested that the apoptotic effect of celecoxib might be achieved through both a COX-2-dependent, and a COX-2-independent mechanism. Conversely, our findings have suggested that celecoxib function in a COX-2-independent pathway to regulate the rate of aging in C. elegans. First, treatments of OSU-03012, a structural analog of celecoxib that exhibits no detectable COX-2 inhibitory activity, can significantly extend the animals mean lifespan by up to 28%. Second, no worm homolog of human COXs has been identified based on sequence similarity. Finally, biochemical studies by our group and others have revealed that both celecoxib and OSU-03012 exhibit a strong inhibitory activity against 3''-phosphoinositide-dependent protein kinase-1 (PDK-1) in vitro, which is a key component of the DAF-2 insulin-like pathway. Given the critical role that the DAF-2 pathway plays in the regulation of longevity, it is possible that celecoxib and its derivatives might extend lifespan by inhibiting the activity of PDK-1. Consistent with the in vitro data, we have found that the FOXO transcription factor, DAF-16, is indeed required for celecoxib or OSU-03012 to extend lifespan; and that these lifespan extensions can be suppressed by PDK-1 gain-of-function mutations. Furthermore, treatments of celecoxib and OSU-03012 enhance the dauer formation of daf-2 mutants and induce nuclear translocation of DAF-16.

Baek HK et al. (2016) J Vet Sci "Neuronal maturation in the hippocampal dentate gyrus via chronic oral administration of Artemisa annua extract is independent of COX-2 signaling pathway in the diet-induced obesity (DIO) mice model."

Baek HK, Choi DH, Park SK, Hwang IK, Kim PS, Song KD, Oh SI, Kim DE, Song JA, Yi SS, Seo HS, Kim SJ

[J Vet Sci, 2016]

Recently, we reported that Artemisia annua (AA) has anti-adipogenic properties in vitro and in vivo. Reduction of adipogenesis by AA treatment may dampen systemic inflammation and protect neurons from cytokine-induced damage. Therefore, the present study was performed to assess whether AA increases neuronal maturation by reducing inflammatory responses, such as those mediated by COX-2. Mice were fed normal chow or a high-fat diet with or without chronic daily oral administration of AA extract (0.2g/10mL/kg) for 4 weeks; then, changes in their hippocampal dentate gyri were measured with immunohistochemistry/immunofluorescence staining for BrdU, DCX, and NeuN, markers of neuronal maturation, and a quantitative Western blot for COX-2 and Iba-1, in order to assess correlations between systemic inflammation (Il-6) and food type. Additionally, we tested the effect of AA in an Alzheimer's disease model of C. elegans and uncovered a potential benefit. In the present study, we show that chronic AA dosing significantly increases neuronal maturation, particularly in the high-fat diet group. This effect was seen in the absence of any changes in COX-2 levels in mice given the same type of food, pointing to the possibility of alternate anti-inflammatory pathways in the stimulation of neurogenesis and neuro-maturation in a background of obesity.

Ching TT et al. (2011) Aging Cell "Celecoxib extends C.elegans lifespan via inhibition of insulin-like signaling but not cyclooxygenase-2 activity."

Hsu AL, Chiang WC, Ching TT, Chen CS

[Aging Cell, 2011]

One goal of aging research is to develop interventions that combat age-related illnesses and slow aging. Although numerous mutations have been shown to achieve this in various model organisms, only a handful of chemicals have been identified to slow aging. Here, we report that celecoxib, a nonsteroidal anti-inflammatory drug widely used to treat pain and inflammation, extends Caenorhabditis elegans lifespan and delays the age-associated physiological changes, such as motor activity decline. Celecoxib also delays the progression of age-related proteotoxicity as well as tumor growth in C. elegans. Celecoxib was originally developed as a potent cyclooxygenase-2 (COX-2) inhibitor. However, the result from a structural-activity analysis demonstrated that the antiaging effect of celecoxib might be independent of its COX-2 inhibitory activity, as analogs of celecoxib that lack COX-2 inhibitory activity produce a similar effect on lifespan. Furthermore, we found that celecoxib acts directly on 3'-phosphoinositide-dependent kinase-1, a component of the insulin/IGF-1 signaling cascade to increase lifespan.

Uram L et al. (2020) Eur J Pharm Sci "Celecoxib substituted biotinylated poly(amidoamine) G3 dendrimer as potential treatment for temozolomide resistant glioma therapy and anti-nematode agent."

Misiorek M, Woowiec S, Filipowicz-Rachwa A, Walajtys-Rode E, Uram L, Markowicz J

[Eur J Pharm Sci, 2020]

Glioblastoma multiforme (GBM) is a one of the most widely diagnosed and difficult to treat type of central nervous system tumors. Resection combined with radiotherapy and temozolomide (TMZ) chemotherapy prolongs patients' survival only for 12 - 15 months after diagnosis. Moreover, many patients develop TMZ resistance, thus important is search for a new therapy regimes including targeted drug delivery. Most types of GBM reveal increased expression of cyclooxygenase-2 (COX-2) and production of prostaglandin E2 (PGE₂), that are considered as valuable therapeutic target. In these studies, the anti-tumor properties of the selective COX-2 inhibitor celecoxib (CXB) and biotinylated third generation of the poly(amidoamine) dendrimer substituted with 31 CXB residues (G3BC31) on TMZ -resistant U-118 MG glioma cell line were examined and compared with the effect of TMZ alone including viability, proliferation, migration and apoptosis, as well as the cellular expression of COX-2, ATP level, and PGE₂ production. Confocal microscopy analysis with the fluorescently labeled G3BC31 analogue has shown that the compound was effectively accumulated in U-118 MG cells in time-dependent manner and its localization was confirmed in lysosomes but not nuclei. G3BC31 reveal much higher cytotoxicity for U-118 MG cells at relatively low concentrations in the range of 2 to 4 M with compared to CBX alone, active at 50-100 M. This was due to induction of apoptosis and inhibition of proliferation and migration. Observed effects were concomitant with reduction of PGE₂ production but independent of COX-2 expression. We suggest that investigated conjugate may be a promising candidate for therapy of TMZ-resistant glioblastoma multiforme, although applicable in local treatment, since our previous study of G3BC31 did not demonstrate selectivity against glioma cells compared to normal human fibroblasts. However, it has to be pointed that in our in vivo studies conducted with model organism, Caenorhabditis elegans indicated high anti-nematode activity of G3BC31 in comparison with CXB alone that confirms of usefulness of that organism for estimation of anti-cancer drug toxicity.

Hegazi NM et al. (2019) Sci Rep "Characterization of phenolic compounds from Eugenia supra-axillaris leaf extract using HPLC-PDA-MS/MS and its antioxidant, anti-inflammatory, antipyretic and pain killing activities in vivo."

Hegazi NM, Dmirieh M, Mahmoud MF, Sobeh M, Wink M, El-Raey MA, Rezq S, El-Shazly AM

[Sci Rep, 2019]

Reactive oxygen species (ROS) are involved in the pathophysiology of several health disorders, among others inflammation. Polyphenols may modulate ROS related disorders. In this work, thirty-two phenolic compounds were tentatively identified in a leaf extract from Eugenia supra-axillaris Spring. ex Mart. using HPLC-MS/MS, five of which were also individually isolated and identified. The extract displayed a substantial in vitro antioxidant potential and was capable of decreasing ROS production and hsp-16.2 expression under oxidative stress conditions in vivo in the Caenorhabditis elegans model. Also, the extract showed higher inhibitory selectivity towards COX-2 than COX-1 in vitro with higher selectivity towards COX-2 than that of diclofenac. The extract also exhibited anti-inflammatory properties: It attenuated the edema thickness in a dose dependent fashion in carrageenan-induced hind-paw odema in rats. In addition, the extract reduced the carrageenan-induced leukocyte migration into the peritoneal cavity at the highest dose. Furthermore, the extract showed antipyretic and analgesic activities in a mouse model. Eugenia supra-axillaris appears to be a promising candidate in treating inflammation, pain and related oxidative stress diseases.

Suthammarak W et al. (2009) J Biol Chem "Complex I function is defective in complex IV-deficient Caenorhabditis elegans."

Yang YY, Sedensky MM, Suthammarak W, Morgan PG

[J Biol Chem, 2009]

Cytochrome c oxidase (COX) is hypothesized to be an important regulator of oxidative phosphorylation. However, no animal phenotypes have been described due to genetic defects in nuclear encoded subunits of COX. We knocked down predicted homologues of COX IV and COX Va in the nematode Caenorhabditis elegans. Animals treated with W09C5.8 (COX IV) or Y37D8A.14 (COX Va) RNAi had shortened lifespans and severe defects in mitochondrial respiratory chain (MRC) function. Amount and activity of complex IV, as well as supercomplexes that included complex IV, were decreased in COX-deficient worms. The formation of supercomplex I:III was not dependent on COX. We found that COX deficiencies decreased intrinsic complex I enzymatic activity, as well as complex I-III enzymatic activity. However, overall amounts of complex I were not decreased in these animals. Surprisingly, intrinsic complex I enzymatic activity is dependent on the presence of complex IV, despite no overall decrease in amount of complex I. Presumably the association of complex I with complex IV within the supercomplex I:III:IV enhances electron flow through complex I. Our results indicate that reduction of a single subunit within the ETC can affect multiple enzymatic steps of electron transfer, including movement within a different protein complex. Patients presenting with multiple defects of electron transport may, in fact, harbor a single genetic defect.

Cerqua C et al. (2018) Biochim Biophys Acta "COX16 is required for assembly of cytochrome c oxidase in human cells and is involved in copper delivery to COX2."

Frasson C, Sacconi S, Cerqua C, Salviati L, Desbats MA, Sartori G, Baldoin MC, Morbidoni V, Doimo M, Basso G, Trevisson E

[Biochim Biophys Acta, 2018]

Cytochrome c oxidase (COX), complex IV of the mitochondrial respiratory chain, is comprised of 14 structural subunits, several prosthetic groups and metal cofactors, among which copper. Its biosynthesis involves a number of ancillary proteins, encoded by the COX-assembly genes that are required for the stabilization and membrane insertion of the nascent polypeptides, the synthesis of the prosthetic groups, and the delivery of the metal cofactors, in particular of copper. Recently, a modular model for COX assembly has been proposed, based on the sequential incorporation of different assembly modules formed by specific subunits. We have cloned and characterized the human homologue of yeast COX16. We show that human COX16 encodes a small mitochondrial transmembrane protein that faces the intermembrane space and is highly expressed in skeletal and cardiac muscle. Its knockdown in C. elegans produces COX deficiency, and its ablation in HEK293 cells impairs COX assembly. Interestingly, COX16 knockout cells retain significant COX activity, suggesting that the function of COX16 is partially redundant. Analysis of steady-state levels of COX subunits and of assembly intermediates by Blue-Native gels shows a pattern similar to that reported in cells lacking COX18, suggesting that COX16 is required for the formation of the COX2 subassembly module. Moreover, COX16 co-immunoprecipitates with COX2. Finally, we found that copper supplementation increases COX activity and restores normal steady state levels of COX subunits in COX16 knockout cells, indicating that, even in the absence of a canonical copper binding motif, COX16 could be involved in copper delivery to COX2.

Chu Q et al. (2019) Int J Biol Macromol "Tetrastigma hemsleyanum tubers polysaccharide ameliorates LPS-induced inflammation in macrophages and Caenorhabditis elegans."

Ye X, Wang Y, Liu Y, Jia R, Chen M, Jiang Y, Chen W, Li Y, Zheng X, Yu X, Chu Q

[Int J Biol Macromol, 2019]

Tetrastigma hemsleyanum Diels et Gilg, a rare herbal plant native to China, possesses high medicinal value. More researches have focused on the flavones in the tubers of Tetrastigma hemsleyanum whereas polysaccharides attract few attentions. In this study, a purified polysaccharide (TTP-1) with the average molecular weights of 478.33kDa was extracted from Tetrastigma hemsleyanum and conducted multiple analysis to identify its structural information. In vitro, TTP-1 could suppress the inflammation, cytotoxicity, genotoxicity, mitochondrial dysfunction and oxidative damage induced by LPS in RAW264.7 cells. Meanwhile, TTP-1 could attenuate inflammation via COX-2, iNOS, MAPKs pathways and ameliorate oxidative damage through Nrf2-Keap1, Sirt1-FoxO1 pathways in RAW264.7 cells. In vivo, the inflammation induced by LPS triggered the suppression of survival ratio and growing development, as well as the restrain of athletic ability in Caenorhabditis elegans. The treatment of TTP-1 reversed the situation induced by LPS. In a word, TTP-1 could ameliorate LPS-induced inflammation both in vitro and in vivo.

Aremu AO et al. (2010) J Ethnopharmacol "In vitro antimicrobial, anthelmintic and cyclooxygenase-inhibitory activities and phytochemical analysis of Leucosidea sericea."

Fawole OA, Finnie JF, Aremu AO, Light ME, Van Staden J, Chukwujekwu JC

[J Ethnopharmacol, 2010]

ETHNOPHARMACOLOGICAL RELEVANCE: Leucosidea sericea is used as a vermifuge and in the treatment of ophthalmia by various tribes in southern African countries. AIM OF THE STUDY: The study aimed at screening leaves and stems of Leucosidea sericea for pharmacological activity and validating the plant's traditional use. A general phytochemical screening was also carried out. MATERIALS AND METHODS: Petroleum ether (PE), dichloromethane (DCM), ethanol (EtOH) and water extracts of the plant parts were investigated for antimicrobial, anthelmintic and cyclooxygenase (COX) inhibitory activities. Gram-positive bacteria (Bacillus subtilis and Staphylococcus aureus), Gram-negative bacteria (Escherichia coli and Klebsiella pneumoniae) and Candida albicans were used for the antimicrobial evaluation. Caenorhabditis elegans was used for the anthelmintic assay using the microdilution technique. Cyclooxygenase-1 and -2 (COX-1 and -2) were used to evaluate the anti-inflammatory potential of the plant extracts. Phytochemical analysis for phenolic compounds, including gallotannins, condensed tannins and flavonoids was done using 50% methanol extracts of the leaves and stems employing spectrophotometric methods. RESULTS: The leaf extracts exhibited broad spectrum antibacterial activity ranging from 0.025 to 6.25mg/ml. The most noteworthy minimum inhibitory concentration (MIC) of 0.025 mg/ml was exhibited by PE and DCM leaf extracts against Bacillus subtilis and Staphylococcus aureus, respectively. In the anthelmintic assay, the best minimum lethal concentration (MLC) value of 0.26 mg/ml was observed for the DCM and EtOH leaf extracts. Both leaf and stem organic solvent extracts exhibited high to moderate inhibition against COX-1 and -2 at a screening concentration of 250 microg/ml. At lower concentrations, the extracts displayed a dose-dependent inhibition, with the lowest IC(50) values of 0.06 microg/ml (COX-1) and 12.66 microg/ml (COX-2) exhibited by the PE extract of the leaves. Generally, the leaf extracts exhibited better pharmacological activities and contained higher amounts of phenolic compounds than the stem extracts. Alkaloids and saponins were only detected in the leaf and stem extracts, respectively. CONCLUSION: The reported results support the local use of Leucosidea sericea against eye infections and as a vermifuge. The pharmacological activities exhibited by the leaf extracts are probably due to their higher phenolic levels.

Li Y et al. (2019) Free Radic Res "Radix Tetrastigma flavonoid ameliorates inflammation and prolongs the lifespan of Caenorhabditis elegans through JNK, p38 and Nrf2 pathways."

Li Y, Ye X, Jiang Y, Liu Y, Chu Q, Zheng X

[Free Radic Res, 2019]

The main flavonoid components of Radix Tetrastigma (RTF) were extracted and identified by UPLC-TOF/MS. In vitro, RTF prevented inflammation in RAW 264.7 cells by suppressing morphological (both cell and nucleus) changes, and decreasing nitric oxide (NO), inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) contents. Exposure to LPS also leads to oxidant damage, and RTF alleviated damage to mitochondria, decreased O₂^- accumulation, and restored the glutathione level. RTF intervention decreased the expression of c-Jun N-terminal kinase (JNK) and p38 phosphorylation, accompanied by downregulation of nuclear factor erythroid 2-related factor 2 (Nrf2) and forkhead box protein O1 (FoxO1). In vivo, aging of Caenorhabditis elegans ( C. elegans ) by paraquat (PQ) was observed through lifespan, lipofuscin, and enzyme analysis. RTF protected against damage in N2 worms but not in daf-16 mutants. Gene expression was further assessed, and p38/PMK-1 and Nrf2/SKN-1 expression in worms was suppressed by PQ, which was reversed by RTF treatment. Together, these results suggested that RTF could help ameliorate inflammation-induced damage through JNK, p38 and Nrf2 pathways.