Biological Research最新文献

Background: Daphnetin has demonstrated various pharmacological activities. The current study evaluated the potential of daphnetin in alleviating unexplained recurrent spontaneous abortion (URSA) and explored underlying mechanisms.

Methods: Mice with URSA were gavaged with 1 mg/kg, 10 mg/kg, and 20 mg/kg of daphnetin, or infected with adeno-associated viruses harboring knockdown of NR4A1 or overexpression of BACH2 before modeling. Human peripheral blood T lymphocytes were induced into CD4⁺ T cells, followed by lentivirus infection and daphnetin treatment. The influence of daphnetin on CD4⁺ T cell viability and Treg and Th17 cell differentiation in cells was analyzed. The concentrations of Treg cells-associated cytokines (TGF-β, IL-10) and Th17 cells-associated cytokines (IL-17, IL-23) in the supernatants of CD4⁺ T cells were assessed. The regulation of NR4A1 on BACH2 was analyzed by ChIP and dual-luciferase assays.

Results: Daphnetin resulted in fewer immature, resorbed, or dead embryos in mice with URSA, with the most pronounced therapeutic effect of 10 mg/kg. Daphnetin attenuated decidual hemorrhage, with a gain in the percentage/number of Treg cells and a loss of the percentage/number of Th17 cells in the spleen and decidual tissues. Daphnetin enhanced the expression of FoxP3, TGF-β, and IL-10, and suppressed the expression of RORγt, IL-17, IL-23, and the contents of TNF-α, IL-6, and IL-1β in CD4⁺ T cells. Overexpression of BACH2 further alleviated URSA deterioration caused by NR4A1 knockdown. Daphnetin mediated the transcriptional activation of BACH2 by upregulating NR4A1.

Conclusions: Upregulation of NR4A1 by daphnetin mediates BACH2 transcription and Th17/Treg cell homeostasis to improve URSA.

Chemical insecticides play a crucial role in securing global food production but have also caused serious environmental and health problems due to their persistence and low target specificity. In response, insect neuropeptides, which are biological macromolecules that act as key regulators of development, metabolism, reproduction, and behavior, are being explored as potential environmentally friendly alternatives for pest control. This review evaluates the prospects and limitations of using neuropeptides and their synthetic analogues as bioinsecticides. We discuss their mechanisms of action, challenges in increasing biostability, and the risks of affecting nontarget species. Strategies to improve, introduce and increase their applicability usage include peptide modification, targeted delivery systems, and the use of molecular techniques such as RNA interference (RNAi) and CRISPR-Cas9 to disrupt neuropeptide signaling pathways with high specificity. Advances in omics technologies and artificial intelligence are accelerating the discovery and design of novel neuropeptide-based agents. Nonetheless, regulatory challenges, high production costs, limited ecological impact data, and the potential for resistance development remain key obstacles. The integration of neuropeptide-based approaches with existing pest control methods, particularly within genetically modified crops and integrated pest management (IPM), could enhance both efficacy and environmental sustainability. Although the direct application of neuropeptides is still limited, targeting neuropeptide-related genes appears to be a promising and practical direction for the future of biological pest control.

Background: Rheumatoid Arthritis (RA) is an autoimmune disease in which HLA-DRB1 alleles encoding the "Shared Epitope" (SE), located in the β-chain of class II HLA-DR molecules, constitute the main genetic risk factor. However, there is scarce information about the role of HLA class I genes (HLA-ABC) in RA susceptibility. The present work aimed to evaluate the distribution of HLA-ABC allele groups in a cohort of Chilean RA patients and healthy subjects (HS), and to explore the influence of HLA-DRB1 SE alleles on this distribution.

Results: 135 RA patients and 122 HS were genotyped for HLA-ABC. The most frequent allele groups were HLA-A*02 (24.0%), HLA-B*39.1 (14.2%), and HLA-C*07 (24.7%) for RA patients, and HLA-A*02 (31.5%), HLA-A*24 (12.8%) and HLA-C*07 (17.7%) for HS. RA patients presented a significantly higher frequency of HLA-C*07 (p = 0.0015) and HLA-B*39.1 (p = 0.037) allele groups compared to HS. After applying the Bonferroni correction, the significant difference remained only for the HLA-C*07 allele group (p = 0.015). In a subset of RA patients (n = 60), positive for HLA-DRB1 SE alleles, the most frequent HLA-ABC allele groups were HLA-A*02 (0-33.3%), HLA-B*39.1 (0-16.7%), and HLA-C*07 (22.2-60.0%), whereas HLA-B*39.2 and HLA-B*52 were the least frequent ones. Overall, HLA-C*07 was the most frequent allele group across RA patients carrying HLA-DRB1 SE alleles.

Conclusions: The HLA-C*07 allele group shows a significantly higher presence in RA patients compared to HS. In contrast, the distribution of most other HLA-ABC allele groups in this cohort displays a similar frequency between RA patients and HS, consistent with data from different populations.

Background: Male subfertility is a global health concern, with spermatogenic dysfunction being a critical cause. Abnormally high level of palmitic acid (PA), a main component of dietary saturated fatty acid, has been reported to be implicated in the spermatogenic dysfunction, accompanied with a decrease of inhibin B (INHB). However, the mechanism underlying PA-induced downregulation of INHB, and the specific function of INHB in the spermatogenesis microenvironment, remain unclear. Since PA is the main substrate of palmitoylation, a common post-translational lipid modification, we investigated the role of palmitoylation in INHB synthetic defects and subsequent dyszoospermia induced by PA in this study.

Methods: Mice were treated with PA for 30 days to establish a high PA model, and a palmitoylation inhibitor 2-bromopalmitate (2BP) was used for spermatogenesis rescuing. Concentrations and motilities of sperms in the cauda epididymides were analyzed, and pathological examinations were performed to assess spermatogenic function. Hormone levels were detected using ELISA. Primary mouse Sertoli cells and TM4 Sertoli cell line were used for in vitro exploration of mechanisms. Acyl biotin exchange assay was used to explore protein palmitoylation. Co-culture of TM4 and GC1 cells was used to explore the effects of Sertoli cell-secreted INHB on spermatogonia in a paracrine manner.

Results: In this study, we found that excessive PA downregulates testicular INHB levels by suppressing expression of its βB subunit (InhβB) in Sertoli cells, with hyper-palmitoylation of the transcription factor SRY-box containing gene 9 (Sox9) serving as a key regulatory node in this process. We further identified the palmitoyl transferase ZDHHC16 as the primary enzyme responsible for PA-induced Sox9 hyper-palmitoylation. Furthermore, INHB was shown to promote spermatogonial proliferation and differentiation in a paracrine manner within the spermatogenic microenvironment, thereby mediating the modulation of spermatogenesis by palmitoylation in Sertoli cells.

Conclusion: Overall, this study demonstrated that INHB synthesis can be suppressed by PA-induced hyper-palmitoylation of Sox9, and decreased secretion of INHB by Sertoli cells directly leads to spermatogenic dysfunction in the testis microenvironment. These findings highlight Sox9 palmitoylation as a candidate target for treatment of dyszoospermia accompanied with dyslipidemia, and underscore the critical role of INHB in regulating spermatogenesis within the testicular microenvironment.

Animal pharming involves producing recombinant protein drugs using transgenic animals. The United States Food and Drug Administration (FDA) has approved certain drugs produced in the milk of transgenic Rabbits. Traditionally, these pharming Rabbits have been developed using conventional transgenic technology, which often results in an unpredictable success rate, uncontrollable transgene insertion sites, varying copy numbers, and generally low recombinant protein yields, typically 1-2 g/L or lower. We hypothesized that utilizing the promoter of a native major milk protein gene to drive transgene expression could significantly enhance yield. To test this, we developed a rabbit line that expresses tdTomato under the control of the CSN2 gene promoter, responsible for encoding β-casein, the most abundant protein in Rabbit milk. We successfully generated knock-in founder Rabbits using CRISPR/Cas9-mediated knock-in technology, augmented by the homology-directed repair (HDR)-promoting small molecule RS-1. These founder Rabbits were able to transmit the knock-in allele to their offspring, producing both heterozygous and homozygous tdTomato knock-in Rabbits. Remarkably, the recombinant protein yield reached 15-20 g/L in the milk of homozygous animals. Our work demonstrates a promising strategy to enhance recombinant protein production in Rabbit pharming.

Epilepsy is a chronic neurological disorder characterized by a propensity for seizures due to an imbalance between excitatory and inhibitory brain activity. This condition also induces neuroinflammation, which contributes to disease progression. Given that hemichannels (HCs) permeabilize the cell membrane of glia playing a critical role in neuroinflammation, we investigated the antiepileptic potential of Boldo (Peumus boldus M.), an endemic Chilean tree containing several bioactive molecules including boldine, a HC inhibitor. Mice were treated with pulverized Boldo leaves, the antiseizure medication valproate, or a combination of both for 5 days. Seizure severity was assessed in a pentylenetetrazole-induced kindling mouse model. Using the dye uptake technique, we evaluated the membrane permeability in hippocampal astrocytes, microglia, and neurons. Additionally, we analyzed astroglial and microglial reactivity and measured levels of pro-inflammatory cytokines (IL-1β, IL6, and TNF-α). Both Boldo and valproate significantly reduced seizure severity. However, distinct mechanisms were observed. Valproate administration increased dye uptake in control animals and enhanced glial reactivity, corroborating its established ability to stimulate hemichannel activity. Conversely, Boldo treatment, either alone or in conjunction with valproate, reduced these parameters, consistent with its HC-blocking properties. Importantly, Boldo was more effective than valproate in reducing plasmatic levels of inflammatory and oxidative stress markers. These findings indicate that Boldo, by inhibiting these HCs, could provide a valuable therapeutic strategy to mitigate neuroinflammation in epilepsy, highlighting the clinical potential of this readily available medicinal herb.

Background: Osteosarcoma (OS) is the most common non-hematogenous primary malignancy in the bone. Due to several origins of OS, 30-40% OS patients would experience recurrence and metastasis, with a 5-year survival rate of 20-30%. Mesenchymal stem cells (MSCs) transform into OS cells during the differentiation into osteoblasts, and circular RNA (circRNA) hsa_circ_0003611 might contribute to the differentiation of MSCs into osteoblasts. However, the role of hsa_circ_0003611 in the transformation of MSCs into OS cells is largely unknown. This study aims to investigate whether hsa_circ_0003611 tunes the transformation of MSCs into OS cells.

Methods: Here, human bone marrow mesenchymal stem cells (hBMSCs) with hsa_circ_0003611 stably silenced was constructed. Moreover, protein-RNA interaction was detected by RNA immunoprecipitation (RIP), and N⁶-methyladenosine (m⁶A) modification of hsa_circ_0003611 was determined using methylated RNA immunoprecipitation (MeRIP).

Results: The present study reveals that hsa_circ_0003611 level is almost absent in OS cells compared to that in osteoblasts and MSCs. Moreover, hsa_circ_0003611 silence enhances the transformation of MSCs into OS cells in vitro and triggered tumorigenicity of MSCs for OS in vivo. Mechanistically, silence of hsa_circ_0003611 promotes the transformation of MSCs into OS cells by activating MYC proto-oncogene, bHLH transcription factor (MYC) via insulin like growth factor 2 mRNA binding protein 3 (IGF2BP3). Moreover, hsa_circ_0003611 silence improves MYC mRNA stability by facilitating the association between IGF2BP3 and MYC mRNA in MSCs. Furthermore, m⁶A modification disrupts the association between hsa_circ_0003611 and IGF2BP3 to enhance the association between IGF2BP3 and MYC mRNA in MSCs.

Conclusion: In summary, these findings highlight the role of hsa_circ_0003611 in the transformation of MSCs into OS cells and provide novel targets and strategies for OS treatment.