Unveiling Cellular Microenvironments with a Near-Infrared Fluorescent Sensor: A Dual-Edge Tool for Cancer Detection and Drug Screening.

Mitochondria and lysosomes are pivotal intracellular organelles, and the injury or dysfunction of these organelles can trigger a range of pathological processes. Early diagnosis and treatment of cancer are of paramount importance due to cancer's status as a leading health threat. This study introduces a novel fluorescent probe, BDHV, for detecting mitochondrial and lysosomal viscosity and pH abnormalities in tumors, facilitating early cancer detection and screening of anticancer drugs. Under acidic conditions, the red fluorescence of the probe gradually increases with increasing viscosity. Conversely, in alkaline environments, an increase in viscosity leads to a decrease in green fluorescence and an increase in red fluorescence. The inclusion of a benzothiazole group endows BDHV with strong dual-targeting capability for mitochondria and lysosomes and without being affected by the mitochondrial membrane potential. Most notably, BDHV has potential applications for early cancer diagnosis and in effectively assessing the efficacy of various anticancer drugs.

Molecular cloning and characterization of BNIP3 and NIX1/2 and their role in DHA-induced mitophagy and apoptosis in grass carp (Ctenopharyngodon idellus) adipocytes.

B-cell lymphoma-2 and adenovirus E1B 19-kDa-interacting protein 3 (BNIP3) and BNIP3 like (BNIP3L or NIX) play a vital role in regulating mitophagy and the intrinsic apoptosis in mammals, but their gene characterizations remain unclear in fish. Herein, bnip3, nix1 and nix2 were isolated and characterized from grass carp (Ctenopharyngodon idellus), which encode peptides of 194, 233 and 222 amino acids, respectively. As typical BH3-only proteins, grass carp BNIP3, NIX1 and NIX2 proteins contain BH3 and C-terminal transmembrane domains for inducing apoptosis. Moreover, the LC3-interacting region motif of BNIP3, NIX1 and NIX2 is also conserved in grass carp. Phylogenetic analyses also demonstrated that nix1 and nix2 may have originated from the genome duplication event. Expression pattern analysis indicated that bnip3, nix1 and nix2 were highest expressed in brain, followed by eye (bnip3) and liver (nix1 and nix2). BNIP3, NIX1 and NIX2 localized to the nucleus and the cytoplasm, with a predominant localization to mitochondria within the cytoplasm. In the present study, we found that 200 µM DHA impaired the mitochondrial function, manifested as the decreased antioxidant ability, cellular ATP content and mitochondrial membrane potential in grass carp adipocytes. In addition, the gene expression and enzyme activities of caspase family were significantly increased in 200 µM DHA group, indicating that adipocyte apoptosis was induced. Meanwhile, DHA increased the gene expression of bnip3, nix1 and nix2 in a dose-dependent manner in grass carp adipocytes. The colocalization of mitochondria and lysosomes was promoted by 200 µM DHA treatment, implying that BNIP3/NIX-related mitophagy was activated in adipocytes. Based on these findings, it can be inferred that BNIP3/NIX-related mitophagy may be involved in the adipocyte apoptosis induced by DHA in grass carp.

Docosahexaenoic acid (DHA) inhibits abdominal fat accumulation by promoting adipocyte apoptosis through PPARγ-LC3-BNIP3 pathway-mediated mitophagy.

Obesity has always been an overwhelming health concern worldwide. Docosahexaenoic acid (DHA) reduces abdominal fat accumulation by inducing adipocyte apoptosis, but the underlying mechanism remains unclear. Mitophagy, the process of maintaining mitochondrial homeostasis, has a double-edged sword effect that positively or negatively regulates apoptosis. In this study, grass carp (Ctenopharyngodon idellus) was used as an animal model to investigate the role of mitophagy in regulating apoptosis and the potential molecular mechanisms for DHA-induced mitophagy in vivo and in vitro. Firstly, we found that DHA induced the intrinsic apoptosis in grass carp adipocytes, accompanying by activating BNIP3/NIX-mediated mitophagy. Then, suppression of mitophagy alleviated apoptosis and eliminated the inhibition of lipid accumulation induced by DHA in vivo and in vitro. Mechanistically, the DHA-induced mitophagy was caused by activating PPARγ and its DNA binding capacity to the LC3 promoter, which promoted the interaction of BNIP3 (rather than NIX) with LC3. However, the inhibition of PPARγ in vitro significantly decreased the expression of autophagy-related genes (P < 0.05), reducing the colocalization of mitochondria and lysosomes while preventing BNIP3/NIX-mediated mitophagy-mediated apoptosis and subsequently alleviating the inhibition of lipid accumulation in adipocytes induced by DHA. For the first time, we demonstrated that DHA activates mitophagy by regulating the PPARγ-LC3-BNIP3 pathway, consequently inducing apoptosis, which decreases adipocytes, inhibiting lipid accumulation in grass carp. These findings provide new insight into the mechanism of DHA-induced apoptosis mediated by mitophagy as the potential therapeutic target of inhibiting abdominal fat accumulation in vertebrates.

Multicolored inorganic electrochromic materials: status, challenge, and prospects.

Against the backdrop of advocacy for green and low-carbon development, electrochromism has attracted academic and industrial attention as an intelligent and energy-saving applied technology due to its optical switching behavior and its special principles of operation. Inorganic electrochromic materials, represented by transition metal oxides, are considered candidates for the next generation of large-scale electrochromic applied technologies due to their excellent stability. However, the limited color diversity and low color purity of these materials greatly restrict their development. Starting from the multicolor properties of inorganic electrochromic materials, this review systematically elaborates on recent progress in the aspects of the intrinsic multicolor of electrochromic materials, and structural multicolor based on the interaction between light and microstructure. Finally, the challenges and opportunities of inorganic electrochromic technology in the field of multicolor are discussed.

DHA induces adipocyte lipolysis through endoplasmic reticulum stress and the cAMP/PKA signaling pathway in grass carp (Ctenopharyngodon idella).

Docosahexaenoic acid (DHA) is a biologically active fatty acid that reduces the accumulation of lipids. However, the molecular mechanism underlying this process, particularly in fish, is not well understood. Recent studies show that endoplasmic reticulum (ER) stress triggers the activation of the unfolded protein response, which has been revealed to play an essential role in lipid metabolism. In this study, we explored the effect of DHA on ER stress and investigated the potential molecular mechanisms underlying DHA-induced adipocyte lipolysis in grass carp (Ctenopharyngodon idella) both in vivo and in vitro. We found that DHA remarkably reduced the triglyceride content, increased the secretion of glycerol, promoted lipolysis in adipocytes and evoked ER stress, whereas inhibiting ER stress using 4-phenyl butyric acid (4-PBA) inhibited the effects of DHA (P < 0.05). These results implied that ER stress potentially participates in DHA-induced adipocyte lipolysis. Additionally, STF-083010, a specific inositol-requiring enzyme 1α (IRE1α)-inhibitor, attenuated the effects of DHA on lipolysis, demonstrating that IRE1α and X-box binding protein 1 potentially participate in DHA-induced lipolysis. DHA also activated the cyclic adenosine monophosphate (cAMP)-dependent protein kinase A (PKA) pathway by increasing the level of cAMP and activating the PKA enzyme (P < 0.05). Nevertheless, H89, a PKA inhibitor, weakened DHA-induced lipolysis by inhibiting the cAMP/PKA signaling pathway. Furthermore, inhibiting ER stress using 4-PBA also inhibited lipolysis and alleviated DHA-induced activation of the cAMP/PKA signaling pathway, suggesting that ER stress may participate in DHA-induced lipolysis through the activation of the cAMP/PKA signaling pathway. Our data illustrate that DHA supplementation can be a promising nutritional strategy for ameliorating lipid accumulation in grass carp. The present study elucidated the molecular mechanism for DHA-induced lipolysis in grass carp adipocytes and emphasized the importance of ER stress and the cAMP/PKA pathway in DHA-induced lipolysis. These results deepen our understanding of ameliorating lipids deposition in freshwater fish by targeting DHA.

Selective Electrochromic Regulation for Near-Infrared and Visible Light via Porous Tungsten Oxide Films with Core/Shell Architecture.

Dual-band electrochromic smart windows have become a research hotspot owing to their unique ability to selectively control near-infrared (NIR) and visible (VIS) light. However, the design and exploitation of dual-band electrochromic films are still an extreme challenge due to the scarcity of relevant high-performance materials. To solve this issue, we here proposed a type of porous WO3 film with nanowires/nanoparticles core/shell architecture as a promising candidate, endowing smart windows with a dual-band electrochromic feature. Moreover, the mechanism of the dual-band electrochromism is illustrated by the response of the transmittance spectra in Li+-based or TBA+-based electrolytes to distinguish the electrochemical behavior and the cyclic voltammetry to determine the degree of diffusion-limited kinetics. Our results indicate that the dual-band electrochromic performance is credited to the progressive electrochemical reduction procedure, in which the capacitive charging process gives rise to NIR regulation and the following ion intercalation contributes to VIS light modulation. Furthermore, we develop a dual-band electrochromic energy storage prototype device utilizing the porous WO3 film. This work describes a judicious strategy for designing dual-band electrochromic films, promoting the evolution of dual-band electrochromic technology.

The endoplasmic reticulum stress and B cell lymphoma-2 related ovarian killer participate in docosahexaenoic acid-induced adipocyte apoptosis in grass carp (Ctenopharyngodon idellus).

Docosahexaenoic acid (DHA) lessens adipose tissue lipid deposition partly by inducing adipocyte apoptosis in grass carp, but the underlying mechanism remains unclear. Endoplasmic reticulum (ER) stress and unfolded protein response (UPR) is the novel pathway for inducing apoptosis. This study aimed to explore the potential role of ER stress in DHA-induced apoptosis in grass carp (Ctenopharyngodon idellus) adipocytes. DHA induced apoptosis by deforming the nuclear envelope, condensing the chromatin, and increasing the expression of apoptosis-related proteins and genes in vivo and in vitro (P < 0.05). However, the ER stress inhibitor, 4-phenylbutyric acid (4-PBA), effectively suppressed DHA-induced apoptosis (P < 0.05), indicating that ER stress mediates DHA-induced adipocyte apoptosis. Furthermore, we observed that 200 µM DHA significantly up-regulates the transcripts of B cell lymphoma-2 (BCL-2) related ovarian killer (BOK) in vitro (P < 0.05). BOK is a pro-apoptotic protein in the BCL-2 family, which governs the mitochondria apoptosis pathway. Hence, we hypothesized that BOK might be an important linker between ER stress and apoptosis. We cloned and identified two grass carp BOK genes, BOKa and BOKb, which encode peptides of 213 and 216 amino acids, respectively. BOKa primarily localizes in ER and mitochondria in the cytoplasm, while BOKb localizes in the nucleus and cytoplasm of grass carp adipocytes. Moreover, 200 µM DHA treatment up-regulated the mRNA expression of BOKa and BOKb, whereas 4-PBA suppressed the DHA-induced expressions. These results raised the possibility that BOK participates in DHA-induced adipocyte apoptosis through ER stress signaling, in line with its localization in ER and mitochondria. Two UPR branches, the inositol-requiring enzyme 1 (IRE1α) and activating transcription factor 6 (ATF6) signaling pathways, are possibly important in DHA-induced adipocyte apoptosis, unlike protein kinase RNA-activated-like ER kinase. The study also emphasized the roles of BOKa and BOKb in IRE1α- and ATF6-mediated apoptosis. This work is the first to elucidate the importance of the ER stress-BOK pathway during adipocyte apoptosis in teleost.

Cultivated fish accumulates excessive inedible fats in the abdominal cavity, thus wasting energy and causing metabolic disease. This study showed that docosahexaenoic acid (DHA) induces apoptosis in grass carp adipocytes through endoplasmic reticulum (ER) stress and unfolded protein response (UPR). Therefore, DHA possibly alleviates lipid accumulation partly by reducing the number of adipocytes. Furthermore, transcriptome analysis demonstrated that B cell lymphoma-2-related ovarian killer (BOK) is probably an important linker between ER stress and apoptosis. We first cloned and identified the grass carp BOK genes, BOKa and BOKb. Next, we preliminarily confirmed that BOKa and BOKb participate in DHA-induced apoptosis mainly through activating transcription factor 6 and inositol-requiring enzyme 1 α signaling pathways, the branches of UPR. This study provides a theoretical foundation for the lipid-reduction role of DHA in teleost.

PKA/ATGL signaling pathway is involved in ER stress-mediated lipolysis in adipocytes of grass carp (Ctenopharyngodon idella).

The relationship between endoplasmic reticulum stress (ER stress) and lipolysis in mammals has been widely studied, but it is relatively scarce in fish. The present study used grass carp Ctenopharyngodon idella as a model to investigate the effect of ER stress on lipolysis in adipocytes of fish. We found that ER stress evoked by tunicamycin (TM) treatment significantly induced lipolysis in adipocytes. Subsequently, in order to further investigate whether protein kinase A (PKA) is involved in ER stress-induced lipolysis, we treated adipocytes with PKA activator forskolin and inhibitor H89. The results showed that the mechanism was related to the activation of PKA, especially the catalytic subunit PRKACBa. Notably, we also found that PKA regulates lipolysis by targeting mRNA level and protein and enzyme activities of adipotriglyceride lipase (ATGL). Taken together, our findings suggest that PKA/ATGL signaling pathway is involved in ER stress-mediated lipolysis of grass carp adipocytes. It provides a theoretical basis for further study on the mechanism of lipolysis in fish and other vertebrates.

Endoplasmic reticulum stress is involved in lipid accumulation induced by oleic acid in adipocytes of grass carp (Ctenopharyngodon idella): focusing on the transcriptional level.

It has been extensively claimed that endoplasmic reticulum stress (ER stress) is related to lipid accumulation in mammals, but little is known in fish. This study aims at elucidating the role of ER stress in mediating lipid accumulation induced by monounsaturated oleic acid (OA) with a focus on the transcriptional level. We treated the adipocytes of grass carp with 200 µM and 400 µM OA, respectively, while the control group was treated with 2% bovine serum albumin (BSA). The results showed that cell viability was significantly improved, while 400 µM OA treatment promoted neutral lipid accumulation along with stimulating ER stress more obviously. Although lipolysis and fatty acid ß-oxidation were activated simultaneously, the primary effect of OA seems to be promotion of lipid accumulation. To further explore whether ER stress affects lipid accumulation, 4-phenyl butyric acid (4-PBA), an effective inhibitor of ER stress, was used to pretreat the cells for 4 h. Unsurprisingly, it was found that the mRNA expressions of genes linked with ER stress were decreased. Intracellular triglyceride (TG) content was also decreased, which was in accordance with the mRNA expressions of adipogenic and lipogenic transcription factors as well as their target genes. Collectively, our data shows that ER stress may take part in OA-induced lipid accumulation in adipocytes via activating adipogenesis and lipogenesis. Based on this, strategies for protecting ER could be used to alleviate excessive accumulation of lipid in grass carp adipose tissue.

Docosahexaenoic acid lessens hepatic lipid accumulation and inflammation via the AMP-activated protein kinase and endoplasmic reticulum stress signaling pathways in grass carp (Ctenopharyngodon idella).

The liver is the primary organ for frontline immune defense and lipid metabolism. Excessive lipid accumulation in the liver severely affects its metabolic homeostasis and causes metabolic diseases. Docosahexaenoic acid (DHA) is known for its beneficial effects on lipid metabolism and anti-inflammation, but its molecular mechanism remains unknown, especially in fish. In this study, we evaluated the protective effects of DHA on hepatic steatosis of grass carp (Ctenopharyngodon idella) in vivo and in vitro and mainly focused on the AMP-activated protein kinase (AMPK) and endoplasmic reticulum stress (ER stress) signaling pathway analysis. Grass carp were fed with purified diets supplemented with 0%, 0.5% and 1% DHA for 8 weeks in vivo. 1% DHA supplementation significantly decreased the liver triglyceride (TG), malondialdehyde (MDA), serum tumor necrosis factor α (TNFα) and nuclear factor kappa B (NFκB) contents. DHA administration suppressed ER stress and decreased the mRNA expressions related to hepatic inflammation and lipogenesis, accompanied by the activation of AMPK. Correspondingly, DHA activated the AMPK signaling pathway, and inhibited palmitic acid (PA)-evoked ER stress and lipid accumulation and inflammation of grass carp hepatocytes in vitro. In contrast, the inhibitor of AMPK (compound C, CC) abrogated the effects of DHA to improve PA-induced liver injury and ER stress. In conclusion, DHA inhibits ER stress in hepatocytes by the activation of AMPK and exerts protective effects on hepatic steatosis in terms of improving antioxidant ability, relieving hepatic inflammation and inhibiting hepatic lipogenesis. Our findings give a theoretical foundation for further elucidation of the beneficial role of DHA in vertebrates.

ATF6-DGAT pathway is involved in TLR7-induced innate immune response in Ctenopharyngodon idellus kidney cells.

DGAT1 and DGAT2 are two acyl-CoA:diacylglycerol O-acyltransferase (DGAT) enzymes that catalyze the final step in triglyceride (TG) synthesis. TGs are the primary constituents of lipid droplets (LDs). Although it has been demonstrated that LDs modulate immune and inflammatory responses in CIK cells, little is known about whether DGAT1 and DGAT2 involve in this process. Firstly, grass carp DGAT2 was isolated and characterized, encoding 361 amino acids, and all DGAT2 proteins in genomic structures are conserved in vertebrates. Then, using TLR7 agonist, we induced LDs accumulation in CIK cells. Only DGAT1b and DGAT2 were upregulated in forming TLR7 agonist induced-LDs. Next, we utilized small-molecule inhibitors of DGAT1 and DGAT2. The results indicated that DGAT1 inactivation attenuated TG content and the relative expressions of IFNα3, NF-κB, IL-1ß, and TNFα genes, whereas DGAT2 inhibition decreased TG content and the relative expressions of MyD88, IRF7, IFNα3, NF-κB, IL-1ß, and TNFα genes, implying that DGAT1-generated LDs and DGAT2-generated LDs contribute to TLR7-induced immune response via different signaling pathways. Finally, inhibiting ATF6 effectively decreased DGAT-generated LDs accumulation and the expression of TLR7 signaling-related genes induced by TLR7 agonist, implying that ATF6 UPR pathway may mediate the role of DGAT-generated LDs in TLR7 signaling. Overall, we demonstrate that DGAT1 and DGAT2-catalyzed TAG synthesis may generate different LDs to provide distinct signaling platforms for innate TLR7 signaling.

Perilipin 1-3 in grass carp Ctenopharyngodon idella: molecular characterization, gene structure, tissue distribution, and mRNA expression in DHA-induced lipid droplet formation in adipocytes.

Perilipin family is the main structural proteins of lipid droplet (LD) that is intracellular neutral lipid store ponds, and regulates LD assembly and formation, and is crucial for lipid metabolism. Here three paralogs of perilipin family were characterized from grass carp and their complete coding sequences (CDS) were obtained, including perilipin1, perilipin2, and perilipin3, coding peptides of 492, 454, and 419 amino acids, respectively. The alignment of the homology of grass carp perilipin deduced amino acid sequences with other teleost species showed that the homology with mammalian was less than 55%. PAT (perilipin) domain in mammalian was also predicted in grass carp perilipin 1-3 proteins. Genomic organization analysis revealed that grass carp perilipin1 contained 6 coding exons, while both perilipin2 and perilipin3 consisted of 7 coding exons. The mRNA encoding three paralogs were expressed in a wide range of tissues; perilipin1-3 were primarily expressed in adipose tissue and liver; besides, perilipin3 was also highly expressed in the heart. In vitro, 200 µM DHA increased the proportion of smaller lipid droplets effectively in fully differentiated adipocytes of grass carp. The mRNA expression of perilipin1, perilipin2, and perilipin3 was significantly increased in the adipocytes treated with DHA (P < 0.05, P < 0.01). The same responses of different paralogs in the adipocytes during DHA treatment suggest that they might play synergistic roles in the formation of LDs.

cAMP-dependent protein kinase A in grass carp Ctenopharyngodon idella: Molecular characterization, gene structure, tissue distribution and mRNA expression in endoplasmic reticulum stress-induced adipocyte lipolysis.

Protein kinase A (PKA), one of the most widely studied protein kinases, has many functions in cells, including regulating the metabolism of sugar and lipid. Here we identified nine isoforms of the PKA family in grass carp Ctenopharyngodon idella and obtained their complete coding sequences (CDS), including PRKACAa, PRKACAb, PRKACBa, PRKACBb, PRKAR1A, PRKAR1B, PRKAR2Aa, PRKAR2Ab and PRKAR2B, and PRKACA, PRKACB and PRKAR2A, which may experience fish-specific genome duplication. Sequence analysis showed that the predicted protein structures of PKA gene family members in grass carp were different. Grass carp PRKACAa, PRKACAb, PRKACBa, and PRKACBb contained serine/threonine protein kinases, while PRKAR1A, PRKAR1B, PRKAR2Aa, PRKAR2Ab and PRKAR2B contained two cyclic nucleotide-monophosphate binding domains. PRKACAa, PRKACBa, PRKACBb, PRKAR1A, PRKAR1B and PRKAR2Aa contained 10 coding exons, while PRKACAb and PRKAR2Ab consisted of 12 coding exons and 5 coding exons, respectively. The messenger RNA (mRNA) of the nine PKA isoforms was detected in a wide range of tissues, but their abundance showed tissue-dependent expression patterns. In tunicamycin-induced adipocyte lipolysis, only the mRNA levels of PRKACAb and PRKACBa showed a significant increase in adipocyte (p < .05), indicating that nine PKA isoforms may serve somewhat different roles in endoplasmic reticulum (ER) stress-mediated lipolysis in fish. These results suggested that nine grass carp PKA isoforms may play different roles in tissues, and their expression levels were differently modulated by ER stress in adipocyte.

Nuclear factor-κB subunit p65 is involved in lipopolysaccharide-induced lipid accumulation via regulating DGAT1b in Ctenopharyngodon idellus kidney cells.

Lipopolysaccharide (LPS) can promote the accumulation of triglycerides (TGs) in CIK (Ctenopharyngodon idellus kidney) cells, but the underlying mechanism is unclear. In this study, two genes involved TG synthesis, DGAT1a and DGAT1b, were isolated and characterized from grass carp Ctenopharyngodon idella, which encode peptides of 498 and 501 amino acids, respectively. Phylogenetic and synteny analyses indicated that DGAT1a and DGAT1b could have originated from the teleost-specific genome duplication event. Analysis of the exon-intron structures clarified that genomic structures of all DGAT1 proteins are conserved in vertebrates. DGAT1a mRNA was highly expressed in gut, adipose tissue and heart, while DGAT1b mRNA was highly expressed in liver and kidney. After LPS treatment, only expression of DGAT1b was up-regulated and knockdown of DGAT1b reduced the content of TG, suggesting that DGAT1b is involved in LPS-induced lipid accumulation. To explore the mechanism underlying the transcriptional regulation of DGAT1b in response to LPS, we cloned DGAT1b promoter sequence. Its promoter sequence consists of IRF7, RelA (p65) and RelB binding elements. Dual luciferase assay and q-PCR suggested that the promoter of DGAT1b can be activated by the overexpression of p65, but cannot be triggered by IRF7 and RelB. Mutational analysis shows that the potential p65 binding sites may locate in the region -111/-100 bp of the DGAT1b promoter. These results indicated that DGAT1b is the target gene of NF-κB p65. Finally, inhibiting p65 effectively decreased LPS-induced lipid accumulation. Taken together, we demonstrate that NF-κB p65 takes part in the lipid accumulation by regulating DGAT1b-induced TG synthesis in LPS signalling in CIK cells. The finding that NF-κB p65 links LPS signalling and TG synthesis adds to our growing appreciation of the interplay between immunity and lipid metabolism in fish.

Glycogen synthase kinase-3ß (GSK-3ß) of grass carp (Ctenopharyngodon idella): Synteny, structure, tissue distribution and expression in oleic acid (OA)-induced adipocytes and hepatocytes.

Glycogen synthase kinase-3ß (GSK-3ß) plays a crucial role in regulating lipid metabolism, endoplasmic reticulum (ER) stress and apoptosis in mammals, however, its gene structure and function is little known about in fish. Here, its two paralogs, grass carp (Ctenopharyngodon idella) GSK-3ß1 and GSK-3ß2 were isolated and characterized, encoding peptides of 421 and 457 amino acids, respectively. These two paralogs may have originated from the teleost-specific genome duplication (TSGD) event. Alignment of grass carp GSK-3ß deduced amino acid sequences with select teleost species showed that the protein is conserved. However, two paralogs of the GSK-3ß had great variation in gene structure: the GSK-3ß1 contained eleven exons while the GSK-3ß2 contained nine exons. The two paralogs were expressed in a wide range of tissues, GSK-3ß1 was most expressed in adipose tissue, GSK-3ß2 was most expressed in liver. In OA-induced adipocytes and hepatocytes, we found that GSK-3ß1 mRNA expression was significantly increased only in adipocytes, while the mRNA expression of GSK-3ß2 was dramatically increased both in adipocytes and hepatocytes. These results provide evidence that the GSK-3ß may participate in the process of lipid accumulation in OA-induced adipocytes and hepatocytes of grass carp.

Greater potency of adipocytes compared with preadipocytes under lipopolysaccharide exposure in grass carp Ctenopharyngodon idella.

Excessive body fat is a chronic inflammatory disorder. In this process, white adipose tissue (WAT) performs immune activities because of the dysregulated expression of adipokines. Excessive fat is accumulated in farmed fish, thereby threatening fish health. Studies have shown that adipose tissue is also an active immune organ in fish, capable of participating in and influencing immune responses. Adipocytes are the main cellular component of adipose tissue; however, little is known about the relationship between adipocyte and inflammation in fish. In this study, we analyzed transcriptome changes during adipogenesis in the primary culture of grass carp adipocytes using bioinformatics. The results showed that inflammatory signaling pathway may be activated during grass carp adipocyte differentiation, such as NFκB signaling pathway, Toll-like receptor signaling pathway and Adipocytokine signaling pathway, indicating that grass carp adipocytes have immune activities. Exposure to LPS induced expression of adipokines genes in adipocytes and preadipocytes, including NF-kB, IL-6, MCP-1 and TNFα, suggesting that preadipocytes and adipocytes both have immune response and the immune activity is conserved in vertebrates white adipocytes. Further study found that these immune marker genes were higher expressed in adipocytes compared with preadipocytes in LPS-induced inflammation. In summary, adipocyte should be considered as an active immune site in fish. Adipocytes have greater potency compared with preadipocytes in LPS-induced inflammation. This study indicated that adipocytes and preadipocytes may have different contribution in inflammation.