Jianjie Chen

Effects of fluoride on liver apoptosis and Bcl-2, Bax protein expression in freshwater teleost, Cyprinus carpio

Fish take up fluoride directly from water and are the target organisms for fluoride pollution in the aquatic ecosystems. This study was conducted to evaluate oxidative stress, histopathological changes, apoptosis and Bcl-2, Bax expression in the livers of the common carp (Cyprinus carpio) chronically exposed to fluoride. Our results showed that after 90 d of exposure, the inhibition of SOD, GSH activities and a dose-dependent stimulation of MDA levels in the liver tissues indicated that fluoride caused oxidative stress in the fish. Microscopic examinations showed that damages to the liver tissues and cell organelles in the liver tissues increased with exposure concentration. A positive correlation was observed between the apoptosis index and fluoride levels in the livers (r=0.995). There was a negative correlation between the fluoride concentration of water and the expression of Bcl-2, Bcl-2/Bax (r=-0.98, r=-0.96). A positive correlation was showed between the fluoride concentration of water and the expression of Bax (r=0.96) after 90 d of exposure. Our results suggested that the common carp could tolerate relatively high levels of fluoride but adverse effects of fluoride occurred in the livers of the fish after 90 d of exposure. The apoptosis of liver cells had an important causative role in the process of fluoride-induced pathological changes of liver.

Tissue distributions of fluoride and its toxicity in the gills of a freshwater teleost, Cyprinus carpio

Fish take up fluoride directly from water and are susceptible to fluoride contamination of their environment. In this study, we examined the tissue distributions of fluoride and its toxicity in the gills of the common carp (Cyprinus carpio) chronically exposed to fluoride. Carp were exposed to a range of aqueous fluoride (35-124 mg/L) and sampled at 30, 60 and 90 days. The accumulation of fluoride in the tissues increased with the level and duration of exposure. Steady state was not achieved under the experimental conditions. The gills accumulated the highest levels of fluoride followed by the liver>brain>kidney>muscle>intestine. A dose-dependent inhibition was observed for the enzyme activities of Na(+)-K(+)-ATPase and Ca(2+)-ATPase in the gills after the fish were exposed for 90 days. Also, accumulation of fluoride was associated with the inhibition of superoxide dismutase (SOD) activities and a dose-dependent stimulation of malondialdehyde (MDA) levels in the gill tissues, suggesting that fluoride promoted oxidative stress in the fish. Microscopic examinations revealed injuries to gill tissues and chloride cells, with the severity of injury increasing with exposure concentration. These results suggest that chronic exposure to elevated concentrations of fluoride may induce toxicity in the common carp.

Protective properties of sesamin against fluoride-induced oxidative stress and apoptosis in kidney of carp (Cyprinus carpio) via JNK signaling pathway.

Sesamin, a major lignan derived from sesame seeds, has been reported to have many benefits and medicinal properties. However, its protective effects against fluoride-induced injury in kidney of fish have not been clarified. Previously we found that fluoride exposure caused damage and apoptosis in the kidneys of the common carp, Cyprinus carpio. In this study, the effects of sesamin on renal oxidative stress and apoptosis in fluoride-exposed fish were determined. The results showed that sesamin alleviated significantly fluoride-induced renal damage and apoptosis of carp in a dose-dependent manner, indicated by the histopathological examination and ultrastructural observation. Moreover, treatment with sesamin also inhibited significantly fluoride-induced remarkable enhancement of reactive oxygen species (ROS) production and oxidative stress, such as the increase of lipid peroxidation level and the depletion of intracellular reduced glutathione (GSH) level in kidney. To explore the underlying mechanisms of sesamin action, we found that activities of caspase-3 were notably inhibited by treatment with sesamin in the kidney of fluoride-exposed fish. Sesamin decreased the levels of p-JNK protein in kidney, which in turn inactivated pro-apoptotic signaling events by restoring the balance between mitochondrial pro- and anti-apoptotic Bcl-2 and Bax proteins and by decreasing the release of mitochondrial cytochrome c in kidney of fluoride-exposed fish. JNK was also involved in the mitochondrial extrinsic apoptotic pathways of sesamin effects against fluoride-induced renal injury by regulating the levels of p-c-Jun, necrosis factor-alpha (TNF-α) and Bak proteins. These findings indicated that sesamin could protect kidney against fluoride-induced apoptosis by the oxidative stress downstream-mediated change in the inactivation of JNK signaling pathway. Taken together, sesamin plays an important role in maintaining renal health and preventing kidney from toxic damage induced by fluoride.

Effects of fluoride on growth, body composition, and serum biochemical profile in a freshwater teleost, Cyprinus carpio.

Fluoride is ubiquitously distributed in natural waters. Elevated fluoride may cause histopathological changes and induce oxidative stress in the gills of the common carp (Cyprinus carpio). The present study further evaluates the effects of fluoride on growth performance, body composition, and biochemical measurements of C. carpio. The results showed that food intake, growth, serum osmolality, body composition, and biochemical measures in the blood were affected by fluoride. Weight gain rate and specific growth rate in the exposed fish decreased compared with those of the control fish. Levels of crude protein and crude lipids were reduced in the fluoride-exposed fish. The major ion levels in the sera of fluoride-exposed fish were severely disturbed, resulting in a lower osmolality. All the biochemical parameters measured in the blood were affected by the exposure to fluoride. Total protein, albumin, globulin, and glucose in fish exposed to 63.6 mg/L, 77.7 mg/L, and 124.4 mg/L were lower than those in the control fish. Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were markedly increased in exposed fish compared with control. Taken together, the data showed that exposure to fluoride caused a suite of detrimental effects in C. carpio, which might lead to a decrease in growth and food utilization efficiency. The results of the present study indicated that high levels of fluoride could pose a threat to carp in the field.

Fluoride-induced apoptosis and expressions of caspase proteins in the kidney of carp (Cyprinus carpio).

The study was conducted to investigate oxidative stress, apoptosis, and protein expressions of caspase‐3, 8, and 9 in kidney of the carp juveniles exposed to 0, 40, 80, 120, and 160 mg L−1 of fluoride (in the form of NaF) for 90 days. The results showed that dose‐ and time‐dependent decrease of SOD and GSH and dose‐ and time‐dependent increase of MDA were observed in the carp juveniles, which suggested that fluoride induced oxidative damage accompanied with morphological changes and significant apoptosis in fish exposed to fluoride, especially in the higher doses. Fluoride exposure also significantly elevated the protein expressions of caspase‐3, 8, and 9. In conclusion, these results indicate that chronic exposure to fluoride causes oxidative stress, damages the kidney structure, and results in renal apoptosis by caspase‐dependent pathway.

Effects of sodium fluoride on MAPKs signaling pathway in the gills of a freshwater teleost, Cyprinus carpio.

Exposure to elevated levels of fluoride can cause a variety of adverse effects in fish. Previously we showed that fluoride causes injuries and apoptosis in the gills of Cyprinus carpio. In this study, the effects of fluoride on caspase-3 activity and on accumulation of proteins in the MAPKs pathways were evaluated using Western blotting and immunohistochemistry methods in vivo and in vitro. In vivo experiments showed that the caspase-3 activity increased with fluoride exposure level in a dose-dependent pattern Western blotting and immunohistochemistry results indicated that ERK relative activation tended to decrease as a function of fluoride exposure concentration. In contrast, relative activation of JNK increased with fluoride exposure level. Fluoride exposure did not appear to affect p38 activation. Furthermore, pretreatment of branchial cells with MAPK-specific inhibitors effectively prevented JNK induction and ERK inhibition, respectively, as well as reversed caspase-3 activity in fluoride-treated branchial cells. Our results indicate that activation of JNK and inactivation of ERK were caused by increased ROS and decreased antioxidant capacity in the gills of chronically exposed C. carpio described previously, which eventually caused the observed apoptosis in the fluoride-exposed gills and cells in C. carpio. JNK activation and ERK inactivation mechanism play a crucial role in gill impairment induced by chronic fluorosis. These findings contribute to a better understanding of the initial molecular and cellular events in the gill of fish chronically exposed to fluoride.

Waterborne fluoride exposure changed the structure and the expressions of steroidogenic-related genes in gonads of adult zebrafish (Danio rerio)

Excessive fluoride in natural water ecosystem has been demonstrated to have adverse effects on reproductive system in humans and mammals, while the most vulnerable aquatic organisms were ignored. In this study, the effects of waterborne fluoride on growth performance, sex steroid hormone, histological structure, and the transcriptional profiles of sex steroid related genes were examined in both female and male zebrafish exposed to different concentrations of 0.79, 18.60, 36.83 mg L(-1) of fluoride for 30 and 60 d to investigate the effects of fluoride on reproductive system and the underlying toxic mechanisms caused by fluoride. The results showed that the body weight was remarkably decreased, the structure of ovary and testis were serious injured, and the T and E2 levels were significantly reduced in male zebrafish. The transcriptional profiles of steroidogenic related genes displayed phenomenal alterations, the expressions of pgr and cyp19a1a were significantly up-regulated, while the transcriptional levels of er, ar and hsd3β were decreased both in the ovary and testis, and hsd17β8 were down-regulated just in males. Taken together, these results demonstrated that fluoride could significantly inhibit the growth of zebrafish, and notably affect the reproductive system in both sex zebrafish by impairing the structure of ovary and testis, altering steroid hormone levels and steroidogenic genes expression related to the synthesis of sex hormones in zebrafish.

Expression of ERK and p-ERK proteins of ERK signaling pathway in the kidneys of fluoride-exposed carp (Cyprinus carpio).

Chronic exposure to fluoride can result in a variety of adverse effects in fish. Previously we indicated that high fluoride caused damage and apoptosis in the kidneys of the common carp, Cyprinus carpio. In this study, the effects of fluoride on the expression and localization of ERK and p-ERK proteins in the ERK signaling pathway were determined using Western blotting and immunohistochemical methods in the kidneys of carp exposed to 0, 40, 80, 120mg/L fluoride, respectively. Western blotting analysis found that compared with the controls, the levels of ERK1 and ERK2 proteins were relatively unchanged in fluoride-exposed fish, while p-ERK1 and p-ERK2 protein levels decreased significantly with the increased fluoride concentrations. The immunohistochemical analysis found the proteins of ERK and p-ERK were predominantly localized in the cytoplasm of epithelial cells in the renal tubules of C. carpio. Compared with the control group, the levels of ERK protein were relatively constant, yet the levels of p-ERK protein and p-ERK/ERK ratio were reduced with fluoride exposure dose. These findings indicate that the renal damage in carp exposed to fluoride is mediated via the ERK pathway. Fluoride exposure could inactivate ERK, inhibit the expression of p-ERK protein, and induce renal damage in C. carpio.

Molecular cloning and expression analysis of FTZ-F1 in the GIFT tilapia,Oreochromis niloticus

The FTZ-F1 genes encode orphan receptors of the nuclear receptor superfamily and in mammals have been found to play important roles in the proper development of the adrenal-gonadal axis and sex-determination. We isolated the homologue of FTZ-F1 in genetically improved farmed tilapia (gfFTZ-F1). The full-length cDNA was isolated from the ovary, which included an open reading frame encoding a predicted protein of 486 amino acids. Sequence, tissue distribution and phylogenic analysis of the FTZ-F1 showed that the gfFTZ-F1 belonged to SF-1/Ad4BP group and that gfFTZ-F1 transcripts were only expressed in the gonads and kidney but not in other tissues. Likewise our data suggests that the gfFTZ-F1 gene may share similar functions with other fish and mammalian counterparts, though further study is needed to make any definitive conclusions.