Journal of Molecular Histology最新文献

Obesity is one of the major risk factor for infertility since it causes decreased quality and quantity of gametes and a disrupted uterine environment which might result in miscarriage, stillbirth, and fetal abnormal growth. Obesity induces oxidative stress which is strongly associated with infertility. The clearing of oxidative stress by autophagy is maintained through the p62/ Keap1/Nrf2 pathway. In this pathway, oxidative stress induces p62 for binding to Keap1, thereby Keap1 cannot bind to the Nrf2 transcription factor. Then, Nrf2 translocates into the nucleus and initiates antioxidant-related gene expression. While p62, bound to Keap1, acts as an adaptor protein between autophagosome and damaged substrates which needs to be degraded for homeostasis. Up to date, obesity is strongly linked to abnormal autophagy activity. However, p62 protein expression has not been investigated in the obese ovary, testis, and uterus in detail. Thus, in the present study, we aimed to evaluate the effects of a high-fat diet (HFD)-induced obesity on p62 protein levels of the ovary, testis, and uterus in mice. Our results demonstrated that the p62 expression level was significantly altered by HFD in uterine glands, epithelium, myometrium, and stroma, and in the ovarian corpus luteum, testicular spermatogonium and spermatocytes.

Background

Acute respiratory distress syndrome (ARDS) is a life-threatening condition associated with the inflammatory activation of alveolar macrophages. Here, we examined the role of circVAPA in regulating inflammasome activation and macrophage inflammatory polarization in an ARDS model.

Methods

circVAPA expression levels were analyzed in macrophages isolated from healthy controls and patients with ARDS. In vitro cell models of mouse alveolar macrophages and an in vivo mouse ARDS model were established through Lipopolysaccharide (LPS) stimulation. The effects of circVAPA knockdown on macrophage inflammatory polarization, inflammasome activation, and pulmonary tissue damage were investigated in both cell and animal models. The interaction between circVAPA and downstream factors was verified through a luciferase reporter assay and by silencing circVAPA.

Results

circVAPA upregulation in alveolar macrophages was associated with the inflammation in ARDS patients. circVAPA was also upregulated in LPS-stimulated mouse alveolar macrophages (MH-S cells). Additionally, circVAPA knockdown attenuated the inflammatory activation of MH-S cells and reduced the expression of pyroptosis-related proteins. circVAPA silencing also mitigated the inflammatory effects of LPS-stimulated MH-S cells on lung epithelial cells (MLE-12), and alleviated the inflammatory damage in the pulmonary tissue of ARDS mouse model. We further showed that miR-212-3p/Sirt1 axis mediated the functional role of circVAPA in the inflammatory polarization of MH-S cells.

Conclusion

Our data suggest that circVAPA promotes inflammasome activity and macrophage inflammation by modulating miR-212-3p/Sirt1 axis in ARDS. Targeting circVAPA may be employed to suppress the inflammatory activation of alveolar macrophages in ARDS.

Silver nanoparticles (AgNP) exhibit significant cytotoxicity against MKN45 cells (IC50: 105.5 µg/mL). In vivo, AgNP at 150 mg/kg induces necrosis, reduces proliferation, and alters gene expression, presenting a promising gastric cancer treatment strategy. Gastric cancer is the second leading cause of death from cancer worldwide. In this study, the anticancer effect of silver nanoparticles (AgNP) was evaluated in both In vitro and In vivo. First, an MTT assay was employed to estimate the cytotoxicity of AgNP. Next, the obtained IC50s were used as the main doses that were administrated. Regarding In Vitro, MKN45 cells were applied to induce tumor, and AgNP was administrated to mice at doses of 75 and 150 mg/kg for 28 days twice a week in treatment groups post-induction of cancer. After 28 days, the expressions of the BAX, BCL2, and CXCR1 genes were evaluated. An immunohistochemical examination of CD34 and Ki67 markers and tissue absorption of silver nanoparticles were also performed. Our MTT assay results showed that AgNP’s IC50 after 8, 24, and 48 h were 105.5, 70.8, and 22.4 µg/mL, respectively. In addition, the mean survival probability in the treatment groups was more than 25 days. It seemed that the effectiveness of the concentration of 150 mg/kg of silver nanoparticles had caused a significant amount of necrosis in the tumor cells. In addition, the proliferation rate was decreased significantly in the 150 mg/kg group, and the expression of CD34 and Ki67 markers was reduced significantly. However, the expression of BAX and BCL2 genes was increased in the treatment groups. So, as it was shown in this research in both In vitro and In vivo aspects, it seems that the administration of silver nanoparticles can represent a promising strategy in the treatment of gastric cancer.

To explore the impact of amygdalin on the proliferation, migration, and invasion of human endometrial stromal cells (HESCs) and the possible underlying mechanism. HESCs were incubated with 50, 100, and 200 µg/mL of amygdalin. The malignant activities of HESCs were analyzed by functional experiments. The activation of the Wnt/β-catenin signaling was tested using TOP/FOPFlash. The mRNA expressions of genes were validated by qRT-PCR. The endometriosis (EMS) mouse model was induced and the impact of amygdalin on the growth of ectopic endometrial lesions were assessed. It was observed that amygdalin markedly lessened the malignant activities of HESCs in a dose-dependent way (p < 0.05). Amygdalin dose-dependently declined the activation of TOPFlash and mRNA levels of β-catenin, cyclinD1 and c-Myc in HESCs (p < 0.05). Additionally, the increasing dose of amygdalin progressively inhibited the growth of ectopic endometrial lesions in EMS mouse model (p < 0.05). We reached a conclusion that amygdalin could inhibit the malignant activities of HESCs and alleviate EMS, which was related to Wnt/β-catenin signaling activation.

Hypertension alters tooth formation and Atenolol reduces the blood pressure of spontaneously hypertensive rats (SHR) during pregnancy and lactation, and as demonstrated before, increases the microhardness of the SHR offspring’s teeth. MMP-9 is overexpressed in different tissues of hypertensive animals and treatment of hypertension substances can reverse this alteration. We hypothesize hypertension alters the expression of MMP-9 in dental structures of SHR offspring and that treating female SHR with atenolol prevents this alteration. This study aimed to evaluate the expression of matrix metalloproteinase (MMP-9) in incisor teeth (IT) in male offspring of SHR (30 days old) treated or untreated with Atenolol during pregnancy and lactation. MMP-9 expression was evaluated in ameloblasts (AM), enamel matrix (EM), odontoblasts (OD), and predentin (PD) of IT through immunohistochemical reactions (immunoperoxidase). Data were analyzed by Shapiro-Wilk and Kruskal-Wallis (p < 0.05), with Dunn post-test. Histological differences were not observed between IT tissues of SHR and normotensive Wistar rats. For the first time, our data showed that MMP-9 expression in specific dental structures is not altered in SHR. Atenolol treatment increased MMP-9 immunostaining in EM of Wistar rat, however, Atenolol did not alter MMP-9 in the IT tissues of SHR. Our results demonstrated that MMP-9 expression in dental tissues is not affected by hypertension or atenolol treatment in dental tissues. If confirmed in humans, the results obtained in this study will corroborate the suggestion that MMP-9 is not a viable therapeutic target for the treatment of dental alterations associated with maternal hypertension.

Aim

The interaction of MerTK, which negatively regulates immune responses, with its ligand Pros1 contributes to the resolution of apoptosis and inflammation, participating in the healing process of tissues. The levels of MerTK and Pros1, intensely expressed in macrophages (Mϕs), are affected by sex hormones. The expression levels of these proteins in Mϕs, which have a role in corpus luteum (CL) development or regression and folliculogenesis, were investigated in this study since their expressions have not been evaluated in pregnant mouse ovaries.

Method

We analyzed mouse ovaries from non-pregnant mice at estrus and gestation days 5, 8, and 15 (each n:10). We used qPCR to evaluate Mertk and Pros1 mRNA levels and assessed their protein expression and localization using immunohistochemistry and double immunofluorescence staining for co-localization.

Results

Mertk and Pros1 mRNA and protein levels significantly increased in GD15. MerTK and Pros1 protein levels in mouse CL on GD15 were significantly higher than all other groups. MerTK and Pros1 positive Mϕs were observed in CL of GD15 by double immunofluorescence. MerTK protein levels were increased in granulosa cells GD15 of primary and growing follicles.

Conclusion

Our study revealed for the first time that the expression of MerTK and Pros1 was significantly increased in CL at GD15 in mice. These results suggest that increased levels of MerTK andPros1 may enhance their interaction as receptor-ligand binding partners in CL potentially contributing to the balance of apoptosis and inflammation.

Cisplatin resistance is a clinical challenge limiting the treatment of nasopharyngeal carcinoma (NPC). CircRNAs have been evidenced as key molecules involved in tumor advancement and drug resistance. The present study aimed to elucidate the potential biological value of circKRT75 in NPC cisplatin resistance. CircKRT75 levels in NPC clinical samples and parental/resistant cell lines were analyzed based on qRT-PCR. CCK-8 and flow cytometry were adopted to assess the impacts of circKRT75 on the growth viability and apoptotic ability of NPC resistant cells. Meanwhile, western blot was performed to detect changes in the expression of apoptosis-related proteins. Bioinformatics analysis predicted miRNAs and mRNAs downstream of circKRT75, and the interaction between circKRT75 and downstream targets was validated by RNA pull-down, dual-luciferase reporter and rescue experiments. CircKRT75 was notably enhanced in NPC tissues and NPC cisplatin-resistant cells. Functional experiments disclosed that circKRT75 silencing repressed NPC-resistant cell growth and promoted apoptosis. Bioinformatics screening identified that circKRT75 performed as a molecular sponge for miR-659, and CCAR2 was a direct target of miR-659. Further rescue assays confirmed that miR-659 inhibitor restored the inhibitory effect of circKRT75 knockdown on the growth of drug-resistant cells, while CCAR2 silencing could reverse the promotion of NPC cisplatin resistance by circKRT75 upregulation. Additionally, animal experiments revealed that circKRT75 knockdown restrained NPC cisplatin resistance in vivo. CircKRT75 contributed to cisplatin resistance in NPC through miR-659/CCAR2 signaling, which provided a novel perspective and direction to solve the problem of chemoresistance in NPC.

Bone marrow mesenchymal stem cells (BMSCs) indicate a repairing prospect to treat spinal cord injury, a major traumatic disease. This study investigated the repair effect of bone marrow mesenchymal stem cell-derived exosomes (BMSC-Exos) on spinal cord injury. BMSCs were collected to extract BMSC-Exos which were identified by different means. The SCI model of rats was established, the motor behavior was scored by BBB field test, and the spinal cord tissues were separated and stained by HE, Nissl, and Tunel, respectively, as well as analyzed to measure inflammatory and oxidative stress responses. PC12 cells were co-cultured with Exos and analyzed by CCK-8 and flow cytometry to measure cell proliferation and apoptosis. BMSC-Exos improved SCI in rats with the recovery of motor function, alleviation of pathological conditions, and reduction of apoptosis, inflammatory responses, and oxidative stress. BMSC-Exos increased miR-497-5p expression, and miR-497-5p overexpression strengthened the protective effect of BMSC-Exos on SCI. miR-497-5p targeted inactivation of TXNIP/NLRP3 pathway. TXNIP saved the repair effect of miR-497-5p-carrying BMSC-Exos on SCI rats. miR-497-5p-carrying BMSC-Exos alleviated apoptosis and induced proliferation of H₂O₂-treated PC12 cells. BMSC-Exos promote SCI repair via the miR-497-5p/TXNIP/NLRP3 axis, which may be a target for alleviating SCI-associated nerve damage.

An endometrioid carcinogenic pathway of the fallopian tube with possible potential precursors including type II SCOUTs (secretory cell outgrowths) and E-TIN (endometrioid tubal intraepithelial neoplasia) has been recently documented. We report an incidental focus of E-TIN identified in a hysterectomy specimen for Grade 1 endometrioid type endometrial carcinoma. The lesion was present at the fimbriated end of left fallopian tube involving 1 plica. It comprised crowded glandular proliferation with a pseudostratified columnar lining. The cells displayed elongated nuclei with no remarkable nuclear atypia.

Immunohistochemistry showed patchy loss of PAX 2 expression with multifocal aberrant nuclear and cytoplasmic staining for B-catenin. p53 was wild-type and ER was positive.

In view of the co-existing endometrioid type endometrial carcinoma, a possible metastatic spread to the fallopian tube was considered. However, morphologically no obvious nuclear atypia noted, and no associated inflammatory response or desmoplastic stromal reaction identified within the tubal lesion. And on immunostaining, the endometrial tumour was distinct from the tubal lesion. For instance, PTEN was negative/lost in the endometrial tumour but retained in the tubal lesion and B-catenin was membranous in the endometrial tumour but aberrant with multifocal nuclear and cytoplasmic overexpression in the tubal lesion. WT1 was negative in the endometrial tumour but positively expressed by the tubal lesion. All the above findings favoured the possibility of the tubal lesion as being independent of the endometrial primary. In conclusion, we describe an incidental B-catenin aberrant endometrioid type proliferation of the fallopian tube/E-TIN, to raise awareness of such lesions.

Valproic acid (VPA) is a well-known and increasingly documented antiepileptic drug that has been widely used in the treatment of epilepsy and/or epilepsy-related disorders. Prolonged clinical use of VPA has been reported to cause side effects such as nephrotoxicity. Edaravone (EDA) is a powerful free radical scavenger. The aim of the study was to investigate the protective effects of EDA against VPA-induced oxidative renal injury. Four experimental groups were formed by randomly assigning thirty-eight male Sprague Dawley rats. The first group, (Control Group, n = 8), consisted of healthy rats. The second group, (Group II, n = 10), comprised control rats given intraperitoneally EDA (30 mg/kg/day) for seven days. The third group (Group III, n = 10) was administered intraperitoneally only VPA (500 mg/kg/day) for seven days. The last group (Group IV, n = 10) was treated with VPA + EDA for seven days. On the 8th day, kidney tissues were immediately removed from rats. In kidney homogenates, reduced glutathione levels and Na/K⁺-ATPase, paraoxonase1 and prolidase activities were remarkably decreased while catalase, superoxide dismutase, glutathione peroxidase, glutathione reductase, myeloperoxidase, and xanthine oxidase activities and lipid peroxidation, protein carbonyl, advanced oxidized protein products, and hydroxyproline contents were notably elevated in VPA given group. Consistently, administration of EDA decreased renal degenerative changes seen in the kidney tissue of VPA given rats. Treatment with EDA in the VPA group significantly resulted in the recovery of both biochemical and histopathological alterations. As a result, EDA is potentially beneficial to revert oxidative renal damage induced by VPA.