Glucose metabolism disorder related to follicular fluid exosomal miR-122-5p in cumulus cells of endometriosis patients.

In Brief: Elevated expression of miR-122-5p in exosomes in the follicular fluid of patients with endometriosis impairs glucose metabolism in cumulus cells and may further impair oocyte quality. Abstract: Endometriosis (EMs) affects fertility in women of childbearing age in many ways. The underlying mechanisms, including the decrease in oocyte quality, require further investigation. Exosomes, small vesicles responsible for intercellular information exchange, have been found to be involved in many biological events, including follicle development and oocyte meiosis recovery. From the perspective of follicular fluid exosomes, this study aimed to elucidate the mechanisms involved in EMs-related oocyte quality decline. Follicular fluid was collected from three groups of women: the untreated EMs group (EMs_UT), the satisfactorily treated EMs group (EMs_ST), and the control group (Ctrl). Mouse cumulus-oocyte complexes (COCs) were co-cultured with exosomes extracted from follicular fluid during in vitro maturation. Oocyte quality and cumulus cell function were assessed. High-throughput sequencing of miRNA in exosomes was conducted. The function of differentially expressed miRNAs was studied by using SVOG human ovarian granulosa cells transfected with an miRNA mimic and inhibitor. It was found that the follicular fluid exosomes from patients with untreated EMs reduced both the rate of maturation and the quality of mouse oocytes. Overexpression of miR-122-5p in untreated EMs inhibited the translation of key aldolase enzymes related to glucose metabolism and partly impaired glucose metabolism in the cumulus cells of patients with endometriosis. miR-122-5p was also observed to reduce proliferation and increase apoptosis after cell transfection with an miR-122-5p mimic and inhibitor. Further experiments are needed to determine whether there are additional small molecules in the follicular fluid of patients with endometriosis that could be involved in damaging oocyte quality and to identify where harmful substances in follicular fluid exosomes are loaded.

The Association Between Daily Intake of Dietary Supplements and Self-Reported Endometriosis: A NHANES-Based Study.

Dietary supplements are gaining recognition as potential influencers of female reproductive health, but their connection to endometriosis risk remains underexplored. This study addressed this gap, examining the impact of daily dietary supplement intake on the initiation and progression of endometriosis. To explore this, a cross-sectional study was conducted involving 3950 participants representative of the US population from the 1999-2006 National Health and Nutrition Examination Survey (NHANES). Infertility was determined by a question on year-long attempts to become pregnant. Unweighted and weighted multivariate logistic regression analyses assessed the association between dietary supplements and endometriosis risk. Subgroup analysis was conducted based on the participants' body mass index (BMI). The results revealed intriguing patterns. Specifically, higher dietary fiber content (Q4 vs Q1: OR = 0.56, 95% CI = (0.37,0.84), P = 0.0062) and density (Q4 vs Q1: OR = 0.55, 95% CI = (0.38,0.81), P = 0.0035) were linked to reduced risk of endometriosis. Protein content (Q4 vs Q1: OR = 0.47, 95% CI = (0.31,0.74), P = 0.0011) and density (Q4 vs Q1: OR = 0.63, 95% CI = (0.45,0.88), P = 0.0096) similarly exhibited a negative association with endometriosis risk. Interestingly, when stratified by BMI, these effects were pronounced in normal-weight women, whereas they were not evident in the overweight and obese subgroup. Protein content and density showed no significant associations across subpopulations. In conclusion, this study established a negative relationship between dietary fiber and endometriosis, particularly notable in normal-weight women. Future research is essential to validate these findings and establish a causal link between dietary fiber and endometriosis.

A novel p38 MAPK gene in the sea cucumber Apostichopus japonicus (Ajp38) is associated with the immune response to pathogenic challenge.

The p38 mitogen-activated protein kinase (MAPK), an important component of the MAPK signal cascade, is activated by extracellular stimuli, such as environmental stress and pathogenic infection. To clarify the function of p38 MAPKs in echinoderms, we used transcriptome database mining and rapid amplification of cDNA ends (RACE) to identify a novel p38 MAPK gene in the sea cucumber Apostichopus japonicus (here designated Ajp38). The full-length cDNA of Ajp38 was 2231 bp, including an open reading frame encoding 356 amino acid residues. Our sequence analysis indicated that the predicted Ajp38 protein contained the dual phosphorylation site Thr-Gly-Tyr (TGY) and was similar to the p38 homolog in sea urchins. Quantitative real-time PCR analysis showed that Ajp38 was ubiquitously expressed in all examined tissues of healthy adult A. japonicus, with the highest level of expression identified in the coelomocytes. Ajp38 mRNA expression was significantly upregulated in the coelomocytes 4, 12, and 72 h post in vivo infection with Vibrio splendidus. Our results provide more information about the characteristics and immune functions of the p38 homolog in sea cucumbers.

Identification and expression analysis of a TLR11 family gene in the sea urchin Strongylocentrotus intermedius.

In this study, a homolog of the TLR11 family gene from the sea urchin Strongylocentrotus intermedius (denoted as SiTLR11) was cloned and characterized. The full-length cDNA of SiTLR11 was 2096-bp long, which included 43 bp of 5' untranslated region (UTR), 238 bp of 3' UTR, and a putative open reading frame of 1815 bp encoding a polypeptide of 604 amino acid residues. Representative domains such as leucine-rich repeat (LRR) (residues 108-249) and a cytoplasmic Toll-interleukin-1 receptor (TIR) (residues 429-571) domains were detected in the predicted amino acid sequence of SiTLR11. SiTLR11 transcript was widely distributed in all the tested tissues, including intestine, tube feet, gonad, coelomocytes, and peristomial membrane, with the highest expression level in the coelomocytes and peristomial membrane. After the sea urchin was injected with polyinosinic:polycytidylic acid (PolyI:C), the expression level of SiTLR11 in the coelomocytes increased significantly, reaching 1.96-fold the level of the control at 12 h, but decreased to level below that of control at 24 and 48 h. Injection of peptidoglycan (PGN) also led to increased expression of SiTLR11, which peaked at 12 h, yielding an increase of 2.19-fold compared to the control group, and continued to increase at 24 and 48 h. However, almost no differences in immunological activity were found in the groups challenged with lipopolysaccharides (LPS), Zymosan A (ZOA), or Vibrio fortis compared to the control. Taken together, the results strongly suggested that SiTLR11 was functionally involved in the immune response triggered by double-stranded RNA (dsRNA) viruses and Gram-positive bacteria.

cDNA cloning, expression and immune function analysis of a novel Rac1 gene (AjRac1) in the sea cucumber Apostichopus japonicus.

The ras-related C3 botulinum toxin substrate 1 (Rac1) belongs to Ras homolog (Rho) small GTPases subfamily. As an important molecular switch, Rac1 regulates various processes in the cell, especially in cellular immune response. With attempt to clarify characters and functions of Rac1 in sea cucumbers, full length cDNA of a Rac1 homolog in the sea cucumber Apostichopus japonicus (AjRac1) was cloned by transcriptome database mining and rapid amplification of cDNA ends (RACE) techniques. The open reading frame of AjRac1 is 579 bp encoding a protein with a length of 192 aa. Sequence analysis showed that AjRac1 is highly conserved as compared to those from other eukaryotic species. Phylogenetic analysis revealed that amino acid sequence of AjRac1 closely related to those from Strongylocentrotus purpuratus. Results of expression analysis showed that AjRac1 exhibited a relative high expression in blastula stage, adult coelomocytes and respiratory tree in A. japonicus. The transcription of AjRac1 in adult coelomocytes altered significantly at 4 h- and 12 h-after Vibrio splendidus infection, respectively, which indicated that AjRac1 involved in sea cucumber innate immunity. All data presented in this study will deepen our understanding of characterizations and immunological functions of Rac1 in sea cucumbers.

A novel MKK gene (AjMKK3/6) in the sea cucumber Apostichopus japonicus: Identification, characterization and its response to pathogenic challenge.

The mitogen-activated protein kinase kinases (MKKs) are key components of MAP kinase (MAPK) cascades and function as redox-regulated signaling factors in pathological and physiological processes. In this study, we identified a novel MKK3/6 gene in the sea cucumber Apostichopus japonicus (designated as AjMKK3/6) by transcriptome database mining and rapid amplification of cDNA ends (RACE) approaches. Sequence analysis and protein structure prediction showed that AjMKK3/6 is highly conserved as compared to those from other invertebrate and vertebrate species. Molecular phylogeny result revealed that AjMKK3/6 exhibited a closest relationship with that from Strongylocentrotus purpuratus. Quantitative real-time PCR was employed to determine the expression profiles of AjMKK3/6 in healthy adult A. japonicus tissues and in coelomocytes after Vibrio splendidus infection in vivo, respectively. As results shown, AjMKK3/6 was ubiquitously expressed in all examined tissues of healthy adult A. japonicus with a relative expression level from high to low as body wall > tube feet > coelomocytes > respiratory tree > intestine > longitudinal muscle. Significant expression changes of AjMKK3/6 in coelomocytes were observed at 12 h- and 72 h-after V. splendidus infection, respectively. In general, the current study will enrich our knowledge of characterizations and immno-functions of MKK3/6 in sea cucumbers.

[Effects of ursolic acid on liver-protection and bile secretion].

OBJECTIVE: To study the liver-protection and promoting bile secretion of ursolic acid. METHODS: Ursolic acid was administrated to the mice with liver injury induced by CCl4 or APIT. ALT, AST levels in serum were examined. The bile flow rate and the concentration of the main components in the bile of rats administrated with ursolic acid were estimated. RESULTS: Ursolic acid could decrease the serum ALT, AST, SB levels in the mice treated with CCl4 or APIT, promote the secretion of bile and increase the concentration of bilirubin in the bile. CONCLUSION: Ursolic acid exhibited significant liver protection and promoting blie secretion.