Premature ovarian failure (POF) is the leading cause of female infertility, and there is no optimal treatment or medication available currently. For POF, electroacupuncture (EA) has been considered a promising therapeutic approach, but the mechanism for this is not clear. In this study, we explored the effects of EA (CV4, ST36, and SP6) on oxidative stress and intestinal microbiota of high-fat and high-sugar- (HFHS-) induced POF mice. The development of mice follicles was observed by hematoxylin and eosin (HE) staining. The serum levels of estrone (E1), estrogen (E2), estriol (E3), and 21-deoxycortisol (21D) were measured by the HPLC-MS/MS method. The concentrations of Fe2+, superoxide dismutase (SOD), hydroxyl radical (·OH), glutathione (GSH), superoxide anion, and malondialdehyde (MDA) were measured by spectrophotometry. The 16S-rDNA sequencing was used to measure many parameters related to the host gut bacteriome and mycobiome composition, relative abundance, and diversity. mRNA expression levels of ferroptosis-related genes were determined by RT-qPCR. After 4 weeks of EA intervention in POF mice, mature follicles were increased and the levels of the sex hormone were improved. SOD activities, antisuperoxide activities, and GSH increased while MDA, ·OH, and Fe2+ decreased. In addition, EA also altered the intestinal microbiota. These results reveal that EA can effectively inhibit ovarian oxidative stress and the accumulation of Fe2+ in POF mice. It may be that the alteration in the intestinal microbiota is one of the potential mechanisms of EA treatment. These findings suggest that EA has clinical potential as a safe treatment for POF.
BackgroundOvarian cancer stem cells (OCSCs) are the main cause of relapse and drug resistance in patients with ovarian cancer. Anisomycin has been shown to be an effective antitumor agent, but its mechanism of action in ovarian cancer remains elusive.MethodsCD44+/CD133+ human OCSCs were isolated from human ovarian cancer tissues. OCSCs were interfered with using anisomycin and specific small‐interfering RNA (siRNA). Microarray assay, MTT, in vivo tumorigenic experiments, transwell assay, cell cycle assay, colony formation assay, angiogenesis assay, and hematoxylin and eosin staining were used to detect the mechanism of anisomycin with respect to inhibiting the activity of OCSCs. Expression of the NCBP2‐AS2/mitogen‐activated protein kinase kinase (MEK)/extracellular signal‐regulated kinase (ERK)/signal transducer and activator of transcription 3 (STAT3) pathway was examined using western blotting, a quantitative real‐time PCR (RT‐qPCR) and immunofluorescence staining. Bioinformatics analysis was used for predictive analysis of NCBP2‐AS2 expression in urogenital tumors.ResultsMicroarray analysis showed that treatment with anisomycin significantly decreased the expression of antisense RNA NCBP2‐AS2 in OCSCs. In vitro cellular experiments showed that interfering with endogenous antisense RNA NCBP2‐AS2 using siRNA distinctly inhibited the proliferation, migration and angiogenesis of OCSCs, whereas in vivo animal experiments revealed decreased tumorigenesis in nude mice. Moreover, the results of RT‐qPCR and western blotting demonstrated that both anisomycin treatment and NCBP2‐AS2 silencing led to significant reductions in the mRNA and protein expression levels of NCBP2‐AS2, MEK, ERK and STAT3. From a bioinformatic point of view, antisense RNA NCBP2‐AS2 exhibited significantly differential expression between urogenital tumors and normal controls, and a similar expression pattern was found in the genes NCBP2, RPL35A, DNAJC19 and ECE2, which have similarity to NCBP2‐AS2.ConclusionsAnisomycin suppresses the in vivo and in vitro activity of human OCSCs by downregulating the antisense RNA NCBP2‐AS2/MEK/ERK/STAT3 signaling pathway, whereas the antisense RNA NCBP2‐AS2 and genes with similarity have the potential to serve as markers for clinical diagnosis and prognosis of urogenital tumors.
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