The PFOS disturbed immunomodulatory functions via nuclear Factor-κB signaling in liver of zebrafish (Danio rerio).

Excessive perfluorooctane sulfonate (PFOS) in natural water ecosystem has the potential to detrimentally affect immune system, but little is known of such effects or underlying mechanisms in fish. In the present study, we evaluated the effects of PFOS on growth performance, organizational microstructure, activities of immune-related enzymes and expressions of immune-related genes in male zebrafish (Danio rerio) exposed to different concentrations of 0, 0.02, 0.04 and 0.08 mg/L of PFOS for 7, 14, and 21 days or cotreatment with PFOS and PDTC to investigate the effects of PFOS on immune system and the potential toxic mechanisms caused by PFOS. The results indicated that PFOS accumulated in livers after exposure, and remarkably elevations were found in three exposure groups compared with the control group at three stages. The growth of the adult zebrafish in the experiments was significantly inhibited, the microstructures of liver were serious damaged. The ROS levels were remarkably increased. The activities of ACP, AKP, and lysozyme were obviously decreased, while the activities of MPO and NF-κB were significantly increased. The expressions of immune-related mRNA were significantly affected. After co-treatment with PFOS and PDTC, the growth inhibition, the morphological damage, the ROS induction, and the expressions of immune-related mRNA were reversed. Taken together, the results indicated that PFOS can significantly inhibit the growth, disturb the immune system by changing the normal structure of liver, the activities of immune-related enzymes, and a series of gene transcriptions involved in immune regulation in liver of male zebrafish. PFOS-induced pro-inflammatory effect of hepatocytes was observed, and the involvement of NF-κB signaling pathway was participated in its action mechanism. These findings provide further evidence that PFOS interferes with the immune regulation of liver of male zebrafish under in vivo conditions.

[Study on clinical efficacy and mechanism of Qingying Tang for treating psoriatic blood-heat syndrome based on IL-23/Th17].

To observe the clinical curative effect of Qingying Tang in the treatment of psoriatic blood-heat syndrome and explore its intrinsic mechanism.In this study,we collected 72 patients with blood-heat syndrome psoriasis admitted to our dermatology clinic from January 2016 to December 2017 and divided into control group and observation group according to the random number table method,36 cases in each group.The patients in control group were given with Acitretin Capsules orally,10 mg/time,twice a day.The patients in observation group were given with Qingying Tang orally,150 mL/time,twice a day.The treatment period was 12 weeks in both groups.The traditional Chinese medicine(TCM) syndrome scores before and after treatment,psoriasis area and severity index(PASI) score,dermatology life quality index(DLQI) score,and the clinical efficacy of the two groups were compared between the two groups;flow cytometry was used to detect peripheral blood Th17 cell percentages before and after treatment in both groups;serum interleukin(IL)-17,IL-23,IL-22,and IL-21 levels in both groups before and after treatment were measured by ELISA;the expression levels of STAT3 and RORγt before and after treatment in patients were measured by using skin lesion immunohistochemical method.The results showed that the TCM symptoms were improved significantly in both groups(P<0.05),and the effect in observation group was significantly better than that in the control group(P<0.05).PASI and DLQI scores were decreased significantly after treatment in both groups(P<0.05),and the scores in observation group were significantly lower than those in the control group(P<0.05).The curative effect of the observation group was significantly higher than that of the control group(P<0.05).After treatment,the percentage of Th17 cells,as well as IL-17,IL-23,IL-22 and IL-21 levels in peripheral blood were significantly decreased in both groups(P<0.05),and the levels in observation group were significantly lower than those in the control group(P<0.05).The expression levels of STAT3 and RORγt in both groups were significantly lower than those before treatment(P<0.01),and the levels in observation group were significantly lower than those in the control group(P<0.05).All of the results indicted that Qingying Tang can effectively improve the skin lesions and TCM syndrome in patients with psoriasis and blood-heat syndrome,and improve patient health quality,which may be related to regulation of peripheral blood IL-23/Th17.

Activation of tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) receptor gene expression following DNA demethylation in placental choriocarcinoma and transformed cell lines.

We characterised DNA methylation and gene expression of four tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) receptors DR4, DR5, DcR1 and DcR2 in three choriocarcinoma (JAR, JEG-3, BeWo) and two transformed (HTR-8/SVneo and HPT-8) cell lines. DR4 mRNA was detected in JAR, JEG-3, BeWo and HTR-8/SVneo cells, whereas DR5 was present in all detected cells. DcR1 transcripts were expressed only in JAR, JEG-3 and BeWo cells, whereas DcR2 transcripts were detected only in HTR-8/SVneo and HPT-8 cells. Hypermethylated DR4 promoter was observed in JAR, JEG-3, BeWo and HTR-8/SVneo cells, hypermethylated DcR1 promoter in HTR-8/SVneo and HPT-8 cells and hypermethylated DcR2 promoter in JAR, JEG-3 and BeWo cells. Restoration of DR4, DcR1 and DcR2 expression with decreased DNA methylation of these genes was induced by the DNA demethylation agent 5-aza-2'-deoxycytidine (5-aza-CdR) in trophoblast cells, whereas DR5 expression did not exhibit any change. Significant negative correlation between the expression and DNA methylation of these genes was also observed. In all tested cell lines, only HPT-8 demonstrated sensitivity to TRAIL-induced apoptosis. Combined treatment with 5-aza-CdR and TRAIL resulted in apoptosis in JAR, JEG-3, BeWo and HTR-8/SVneo cells but not in HPT-8 cells. The results indicate that DNA methylation is associated with TRAIL receptor expression and might be involved in trophoblast apoptosis.

Interleukin-17A knockout or self-recovery alleviated autoimmune reaction induced by fluoride in mouse testis.

Fluoride (F) is usually treated as a hazardous material, and F-caused public health problem has attracted global attention. Previous studies demonstrate that interleukin-17A (IL-17A) plays a crucial role in F-elicited autoimmune orchitis and self-recovery reverses F-induced testicular toxicity to some extent, but these basic mechanisms remain unclear. Thus, we established a 180 d F exposure model of wild type (WT) mice and IL-17A knockout mice (C57BL/6 J background), and 60 d & 120 d self-recovery model based on F exposure model of WT mice, and used various techniques like qRT-PCR, western blot, immunohistochemistry and ELISA to further explore the mechanism of F-induced autoimmune reaction, the role of IL-17A in it and the reversibility of F-caused toxicity in testis. The results indicated that F exposure for 180 d caused the decreased sperm quality, the damaged testis histopathology, the enhanced mRNA and protein expression levels of inflammatory cytokines, the changes of autoantibody such as the appearance and increased content of anti-testicular autoantibodies in sera and the autoantibody deposition in testis, the alterations of autoimmune related genes containing the decreased mRNA and protein expressions of AIRE and FOXP3 with an increase of MHCII, and the reduced protein expressions of CTLA4, and the activation of IL-17A signaling cascade like the elevated mRNA and protein expressions of IL-17A, Act1, NF-κB, AP-1 and CEBPß, and the increased protein expressions of IL-17RC, with a decrease of IκBα. After IL-17A knockout, 29 of 35 F-induced changes were alleviated. In two self-recovery models, all F-caused differences except fluorine concentration in femur were gradually restored in a time-dependent manner. This study concluded that IL-17A knockout or self-recovery attenuated F-induced testicular injury and decrease of sperm quality through alleviating autoimmune reaction which was involved with the activation of IL-17A pathway, the damage of self-tolerance and the enhancement of antigen presentation.

Self-recovery study of the adverse effects of fluoride on small intestine: Involvement of pyroptosis induced inflammation.

Increasing investigations suggest that fluoride (F) exposure was associated with gastrointestinal diseases, but related literatures were still largely insufficient and the underlying mechanisms have not been fully elucidated. Moreover, previous study in our lab reported F toxicity has the reversible tendency, but it still needs to be further explored. To address this issue, we established a 90 days F exposure and 15 days & 30 days self-recovery mice model, including control and three F groups (25, 50 and 100 mg/L sodium fluoride (NaF)) in each period. The results revealed that after 90 days F exposure, histological structure and ultrastructure of small intestine were markedly disrupted; the value of villus height to crypt depth, and expressions of tight junctions related mRNA and proteins were significantly decreased; intestinal permeability, pro-inflammatory cytokines and pyroptosis related mRNA and proteins were notably increased in duodenum, jejunum and ileum. However, intriguingly, after 30 days recovery period, indices in F groups almost all have recovered towards normalcy. Collectively, this study demonstrated that F exposure could impair the structure and epithelial barrier function of small intestine, leading to the intestinal inflammation, and pyroptosis may contribute to this damage; Furthermore, F toxicity on small intestine is reversible, and could be restored when off the F exposure environment for a certain period of time. Additionally, among the three regions of small intestine, duodenum seems more vulnerable to F exposure than jejunum and ileum.

Fluoride impairs ovary development by affecting oogenesis and inducing oxidative stress and apoptosis in female zebrafish (Danio rerio).

Previous studies have shown that waterborne fluoride exposure has adverse effects on the reproductive system of zebrafish. However, the underlying toxic mechanisms were still not clear. In the present study, female zebrafish were exposed to different concentrations of 0.787 (Control), 18.599, 36.832 mg/L of fluoride for 30 d and 60 d, and the effects of different doses of fluoride on ovary development, reproductive hormones, oogenesis, ROS content, antioxidant levels, and the expression of apoptosis-related genes and proteins in the ovaries of female zebrafish were analyzed. The results showed that ovarian weight and GSI were significantly decreased, FSH, LH and VTG levels were significantly reduced, the transcriptional profiles of oogenesis-related genes (tgfß1, bmp15, gdf9, mprα, mprß, ptg2ß) were remarkably altered, ROS levels was notably increased, the SOD, CAT, GPx activities and GSH content as well as their mRNA expressions were significantly decreased, MDA content was remarkably increased, the expressions of apoptosis-related genes and proteins (caspase3, caspase8, caspase9, Fas-L, Cytochrome C, Bax and Bcl-2) were significantly changed, the ratio of Bax/Bcl-2 protein levels were notably increased. Taken together, this study demonstrated that fluoride exposure significantly affected ovarian development, decreased the reproductive hormones, affected oogenesis, induced oxidative stress, caused apoptosis through both extrinsic and intrinsic pathways in ovary of zebrafish. Indicating that oogenesis, oxidative stress, and apoptosis were responsible for the impairment of ovarian development.

Fluoride Induces Autoimmune Orchitis Involved with Enhanced IL-17A Secretion in Mice Testis.

Fluoride (F) widely exists in the water and food. Recent studies reported that F induced testicular toxicity via inflammation reaction. This study was aimed to explore the mechanism of F-induced inflammation in testis. 100 healthy male mice (BALB/cJ strain) were randomly divided into five groups including: control, experimental autoimmune orchitis (EAO), and three F groups (25, 50, and 100 mg/L sodium fluoride (NaF)). After 150 d, the results showed a significant increase in testicular cytokines levels including of IL-17A, IL-6, IFN-γ, and TNF-α in NaF and EAO groups compared with control group. Interestingly, the presence of specific antisperm autoantibodies in antitesticular autoantibodies and the notable recruitment of immunocyte (T cells and dendritic cells) were also observed in NaF and EAO groups. In addition, findings showed that in NaF and EAO groups macrophages and T cells both significantly secreted IL-17A, and the protein and mRNA levels of cytokines (IL-6 and TGF-ß) were significantly increased. From these results, it can be concluded that autoimmune orchitis and IL-17A are implicated in F-induced testicular inflammation.

Copper caused reproductive endocrine disruption in zebrafish (Danio rerio).

Cu in surface waters has been demonstrated to affect aquatic animals at ecologically relevant concentrations. However, its effects on reproductive endocrine system and the underlying toxicological mechanisms are largely unknown. In this study, zebrafish (Danio rerio) were exposed to 0, 10, 20, 40 µg/L of Cu for 30 days. Growth, gonad histopathology, the hormone levels and the transcriptional profiles of genes in the hypothalamic-pituitary-gonadal (HPG) axis in both sexes were examined. The results indicated that body weight was significantly reduced, the gonadal development was affected, and the levels of E2, T and 11-KT were remarkably disturbed in Cu-exposed fish. Moreover, the expression profiles of steroidogenesis-related genes in gonad (3ßhsd, 17ßhsd, cyp11a1, cyp17, cyp19a, lhr, fshr, hmgra and star) and in brains (ar, cyp19b, erα, er2ß, lhß, fshß, gnrh2, gnrh3, gnrhr1, gnrh2 and gnrh4) displayed alterations after exposure to Cu. These results demonstrated that Cu could suppress the growth of zebrafish and significantly affect the reproductive biology in both sexes by damaging the structure of the gonads, altering the steroid hormone levels and the expressions of endocrine-related genes in HPG of zebrafish. This study suggests that Cu adversely affects the reproductive endocrine system in zebrafish and could pose a potential threat to fish populations inhabiting Cu-contaminated waters.

Fluoride altered rat's blood testis barrier by affecting the F-actin via IL-1α.

Fluoride is known to affect the pro-inflammatory cytokines in the testis. Most of the recent literatures cited that cytokines regulate the blood-testis-barrier (BTB). However, the involvement of cytokines in the fluoride induced toxicity in BTB remains unclear. In order to study this, 60 male Sprague-Dawley (SD) rats were taken and randomly divided into 5 groups which included four fluoride groups exposed to 0, 25, 50, and 100 mg/L NaF in distilled water and one positive control group. On the 29th day of fluoride exposure, the positive control group rats were administered 0.1% CaCl2 solution. Biotin tracer technology and transmission electron microscopy (TEM) analysis were applied to evaluate the function and ultra-structure of BTB. The expression levels of the BTB associated proteins, actin relative protein 3 (Arp3), interleukin-1 alpha (IL-1α), and transforming growth factor beta-3 (TGF-ß3) were determined using Western blotting and Enzyme Linked Immunosorbent Assay (ELISA) respectively, meanwhile the actin filament (F-actin) was detected by fluorescent phalloidin conjugates. Our results revealed that the function and the ultra-structure of BTB in all the fluoride treated groups were damaged with a concomitant significant decreases in basal ectoplasmic specialization (basal ES), associated protein ß-catenin, and F-actin. Moreover, Arp3 levels were significantly increased in 50 and 100 mg/L NaF groups. Meanwhile, IL-1α significantly increased in all the fluoride treated groups. In summary, we concluded that an increase in IL-1α induced by NaF significantly decreased the expression of F-actin and the organization of F-actin highly branched, which might facilitate the BTB's functional and ultra-structural variations.