Rat Uterine Tissue Research Articles

The phytoestrogen, quercetin, in serum, uterus and ovary as a potential treatment for dehydroepiandrosterone-induced polycystic ovary syndrome in the rat.

The exact mechanisms of polycystic ovary syndrome (PCOS) are unknown and there is no effective cure for the disease. The aim of this study was to evaluate the alterations in serum oestradiol and adiponectin levels and in the expression of some important genes in the uterine and ovarian tissues of PCOS rats. The therapeutic effect of quercetin on PCOS was also assessed. Rats were divided into five groups: control, ethanol, quercetin (Q), PCOS and PCOS+Q. After 30 days of oral treatments, the rats' ovaries and uteri were removed and nesfatin-1, aromatase and adipoR1 expressions were quantified with real-time polymerase chain reaction. Serum adiponectin and oestradiol levels were evaluated using enzyme-linked immunosorbent assay technique. The results of this study showed that expression of nesfatin-1 and adipoR1 genes and adiponectin serum levels decreased in the PCOS rats, but aromatase expression and oestradiol level increased. Treatment with quercetin increased the adiponectin level and expression of adipoR1 and nesfatin-1 and decreased both the expression of aromatase and the oestradiol level. Quercetin improved PCOS by phytoestrogenic effects and mimicking oestrogen's function. Quercetin also affects important factors in both the uterus and ovary and could improve the obesity and the diabetic and infertility symptoms of PCOS.

LncRNA/mRNA profiling of endometriosis rat uterine tissues during the implantation window.

Endometriosis is associated with changes in long non-coding RNA (lncRNA) and mRNA expression, but the exact changes during the implantation window are unknown. Therefore, this study aimed to explore the lncRNA and mRNA expression profiles in the uterus of rats with endometriosis during the implantation window. A total of 35 non-pregnant female rats were randomized to the endometriosis (n=13), adipose tissue control (n=8) and blank control (n=14) groups. On the 5th day of pregnancy, the rats were sacrificed to obtain uterine tissues. lncRNA and mRNA were analyzed using gene chips. A total of five differentially expressed lncRNA and four mRNA were validated by reverse transcription-quantitative (RT-q)PCR. Immunohistochemistry and western blotting were used to determine the expression of the ADAM metal-lopeptidase with thrombospondin type 1 motif 7 (Adamts7), tumor protein p53 (Tp53), distal-less homeobox 3 (Dlx3) and pyrimidinergic receptor P2Y6 (P2ry6) proteins. There were 115 upregulated lncRNAs, 51 downregulated lncRNAs, 97 upregulated mRNAs and 85 downregulated mRNAs in the endometriosis group. RT-qPCR confirmed the trends for five lncRNAs and four mRNAs (Adamts7, Tp53, Dlx3 and P2ry6). The relative protein expression levels of Adamts7, P2ry6, Dlx3 and TP53 were significantly different in the endometriosis group (P<0.05 vs. controls). Bioinformatics predicted the co-expression relationship of the selected five lncRNA and four mRNA. Gene ontology and the Kyoto Encyclopedia of Genes and Genomes predicted that Adamts7, P2ry6, Dlx3 and TP53 were involved in endometriosis-related inflammation and reproductive pathways. In conclusion, the changes in the expression of lncRNAs, mRNAs and proteins (Adamts7, P2ry6, Dlx3 and TP53) may possibly affect endometrial receptivity in rats with endometriosis during the implantation window, probably resulting in implantation failure of the embryo.

Effect of electroacupuncture on NF-κB and NLRP3 inflammasome in uterine tissues of rats with primary dysmenorrhea

To observe the effect of electroacupuncture (EA) on nuclear factor kappa B (NF-κB) and nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammasome in uterine tissues of rats with primary dysmenorrhea (PD), thus to explore the possible mechanism of EA for PD. Fifty female Sprague-Dawley (SD) rats were randomly divided into a normal group, a model group, an EA at non-acupoint group, an EA at acupoint group and a Western medicine group, with 10 rats in each group. Except for the normal group, rats in the other four groups were treated with estradiol benzoate combined with oxytocin for 11 d to establish PD rat models. From day 1 of the modeling, rats in the normal group and the model group were only properly grasped without any intervention; Guanyuan (CV 4) and Sanyinjiao (SP 6) were selected for EA treatment in the EA at acupoint group; rats in the EA at non-acupoint group were treated with EA at 5 mm away from the acupoints selected above; rats in the Western medicine group were treated with ibuprofen via gavage. Rats in each group were treated for 10-day successively. On the 11th day, except for the normal group, rats in the other groups were intraperitoneally injected with oxytocin (2 U/rat), and the writhing number within 30 min in each group was compared; the pathological changes in rat uteruses were observed by hematoxylin-eosin (HE) staining, and the pathological damage scores were evaluated. Protein expression levels of NF-κB p65, phospho-NF-κB p65, NLRP3, cysteine aspastic acid-specific protease 1 (caspase-1), interleukin (IL)-1β and IL-18 were detected by Western blot. Compared with the normal group, the writhing number increased significantly (P 0.05) in the Western medicine group; compared with the Western medicine group, the phospho-NF-κB p65/NF-κB p65 ratio, also the NLRP3, IL-1β and IL-18 protein levels of the uterine tissues decreased significantly in the EA at acupoint group (all P 0.05); there were no significant differences between the EA at non-acupoint group and the model group in any indicators (all P>0.05). EA at acupoints significantly improves the pain and pathological damages of PD rats. The mechanism may be related to the reduced uterine inflammation via inhibiting NF-κB phosphorylation and NLRP3 activation in uteruses of PD rats.

Can the negative effects of ketamine abuse on female genital organs be prevented by nimesulide? An experimental study.

The objective of this study is to investigate the effects of nimesulide on ketamine-induced ovarian and uterine toxicity by biochemical and histopathological examinations. Ketamine is an anesthetic agent whose use leads to overproduction of catecholamines. Nimesulide is a cyclooxygenase-2 inhibitor, which has also been reported to exert a significant antioxidant effect. Wistar albino female rats were randomly divided into three groups as follows: ketamine group (60 mg/kg), ketamine (60 mg/kg) + nimesulide (50 mg/kg) group, and a healthy control group. Then, the biochemical levels and histopathological findings in the ovaries and uteri of the rats were examined for malondialdehyde, myeloperoxidase, total glutathione and superoxide dismutase. The study demonstrated that, in the uterine and ovarian tissues of rats that have been administered ketamine, there was a decrease in the levels of total glutathione and superoxide dismutase, while malondialdehyde and myeloperoxidase was increased: however it was observed that these ratios were reversed in the ketamine+nimesulide group. It was also proved that the negative effects of ketamine can be corrected with nimesulide when the myometrial and endometrial thicknesses are compared. Antioxidants such as nimesulide may protect against the damage caused by ketamine to the genital organs in young women.

MicroRNA Dysregulation and Steroid Hormone Receptor Expression in Uterine Tissues of Rats with Endometriosis during the Implantation Window.

Background:Estrogen receptor (ER) and progesterone receptor (PR) are involved in endometriosis, but the involvement of microRNAs (miRNAs) is unknown. The aim of the study was to explore the correlation between miRNA and ER/PR in uterine tissues of rats with endometriosis during the implantation window.Methods:Twenty female Sprague-Dawley rats were randomized in three groups: endometriosis (n = 7), fat tissue control (n = 6), and normal (n = 7) groups. The female rats were mated and sacrificed on day 5 (implantation). Uterine tissues were obtained for hematoxylin-eosin staining, immunohistochemistry, and miRNA expression. Reverse transcription polymerase chain reaction (RT-PCR) was used to validate the expression of rno-miR-29c-3p, rno-miR-34c-5p, rno-miR-141-5p, rno-miR-24-1-5p, and rno-miR-490-5p.Results:The 475 miRNAs were found to differentially express between the endometriosis and normal control groups, with 127 being upregulated and 348 being downregulated. Expression of five miRNAs (rno-miR-29c-3p, rno-miR-34c-5p, rno-miR-141-5p, rno-miR-24-1-5p, and rno-miR-490-5p) were validated by RT-PCR and found to be differentially expressed among the three groups. Expression of ER and PR proteins (immunohistochemistry) in the glandular epithelium and endometrial stroma was significantly different among the three groups (all P < 0.05). Five miRNAs were involved in pathways probably taking part in implantation and fertility.Conclusions:The results suggested that miRNAs, ER, and PR could play important roles in the embryo implantation period of rats with endometriosis. These miRNAs might play a role in endometrial receptivity in endometriosis.

Restoration of uterine redox-balance by methanolic extract of Camellia sinensis in arsenicated rats

Arsenic, an environmental and industrial pollutant causes female reproductive disturbances and female infertility. Several researchers found that the use of Camellia sinensis (CS) (green tea) is effective as an alternative therapeutic strategy in the management of several health ailments. This study explores the role of CS extract against arsenic-induced rat uterine tissue damage. Methanolic extract of CS (10 mg/kg BW) was tested concomitantly in arsenic-treated (10 mg/kg BW) rats for a duration of two-oestrous cycle length (8 days). CS effectively attenuated arsenic-induced antioxidantdepletion and necrosis in uterine tissue. Rats treated with sodium arsenite showed significantlyreduced activities of enzymatic antioxidants like superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) in uterine tissue as evidenced by the results of spectrophotometric and electrozymographic analysis. Co-administration of CS significantly reversed the above oxidative stress markers in uterine tissue along with the histopathological changes in ovarian and uterine tissue. Moreover, an increase in the level of transcription factor NF-κB in the uterine tissue in association with reduced serum levels of vitamin B12 and folic acid were mitigated in arsenic fed rats following CS co-administration.

The effect of repeated cadmium oral exposure on the level of sex hormones, estrous cyclicity, and endometrium morphometry in female rats

Cadmium (Cd) is regarded as a potential endocrine disruptor. However, the exact mechanism by which this metal may interfere with the reproductive system has not yet been elucidated. The present study aimed to investigate the effect of subacute Cd oral administration at daily doses of 0.09, 1.8, and 4.5 mgCd/kg b.w. and the impact of Cd on sex hormones (estradiol (E2) and progesterone (P)) in the plasma and uterus, as well as on estrous cyclicity and histopathological changes in uterine and ovary in female rats after terminating the exposure and after a prolonged observation period (3 months). Moreover, Cd bioaccumulation in the uterine and brain tissue of rats was analyzed. The study revealed that oral Cd exposure induced changes in the plasma levels of steroid hormones: decrease in E2 and increase in P after the highest dose of Cd. Probably, for the first time, it was evidenced that circulation sex hormone disturbances in Cd-exposed rats caused irregular estrous cycle, persisting for 3 months after exposure termination; no alterations in these hormone levels in uterine tissue were noted. Cd did not induce estradiol-like hyperplasia of endometrium, but resulted in endometrial edema irrespective of the dose, and caused damage of the ovaries after the highest dose. In summary, subacute oral exposure of female rats to Cd may lead to long-term disturbances in reproductive system.

The impact of electromagnetic radiation (2.45 GHz, Wi-Fi) on the female reproductive system: The role of vitamin C.

The present study investigated the effects of applied continuous 2.45 GHz electromagnetic radiation (EMR), which might cause physiopathological or morphological changes in the ovarian, fallopian tubal, and uterine tissues of rats. We proposed that the addition of vitamin C (Vit C) may reduce these severe effects. Eighteen female Sprague Dawley rats were randomly divided into three groups with six animals in each: Sham, EMR (EMR, 1 h/day for 30 days), and EMR + Vit C (EMR, 1 h/day for 30 days 250 mg/kg/daily). Total oxidant status (TOS) and oxidative stress index (OSI) levels increased ( p = 0.011 and p = 0.002, respectively) in the EMR-only group in ovarian tissues. In all tissues, TOS and OSI levels significantly decreased in the Vit C-treated group in ovarian, fallopian tubal, and uterine tissues ( p < 0.05). Anti-müllerian hormone levels significantly increased in the EMR group ( p < 0.05) and decreased in the Vit C-treated groups. Estrogen (E2) levels were unchanged in the EMR group, as the differences were not statistically significant. Immunohistochemical examination of the ovaries revealed significant increases in Caspase-3 expressions in the epithelial cells of the EMR group ( p < 0.05). In the EMR group, hyperemia was observed in uterine tissues. Also, Caspase-3 and Caspase-8 were significantly increased in the EMR group ( p < 0.001). Caspase-3 was significantly diminished with Vit C application in the ovarian and uterine tissues ( p < 0.05). Caspase-8 was significantly diminished only in uterine tissues ( p < 0.05). These results indicate that prolonged EMR exposure induced physiopathological changes in the ovarian, fallopian tubal, and uterine tissues due to oxidative damage. Under the conditions of this study, Vit C may have protective effects on female reproductive system against oxidative damage.

Comparison of the Thermal Spread of Three Different Electrosurgical Generators on Rat Uterus: A Preliminary Experimental Study

Background/Aims: The objective of this study was to compare the depth and width of thermal spread caused on rat uterine tissue after application of 3 different electrosurgical generators. Methods: Alsa Excell 350 MCDSe (Unit A), Meditom DT-400P (Unit M), and ERBE Erbotom VIO 300 D (Unit E) electrosurgical units (ESUs) were used. The number of Wistar Hannover rats required to obtain valid results was 10. The primary objective of the study was to compare the 3 ESUs using the same instrument and the same waveform. The secondary objective of the study was to compare the differences between monopolar and bipolar systems of each ESU separately using the same waveform. Results: The thermal spread caused by each ESU using monopolar instruments with continuous and interrupted waveforms was significantly different. Among the 3 devices, Unit A caused the largest thermal uterine tissue spread. On the other hand, Unit E caused the most superficial thermal tissue spread, and the smallest thermal spread among all ESUs. Conclusions: Surgeons should note that different ESUs used with the same power output might create different thermal effects especially in the monopolar configuration within the same waveform, for the same duration, and with the same instrument.

Mild moxibustion at different intervention times on the levels of ET-1 and NO in the uterine tissues of rats with cold-damp coagulation and stagnation type dysmenorrhea

ObjectiveTo observe the differences in analgesic effect of moxibustion at different intervention times on dysmenorrhea rats and explore its effect mechanism. MethodsForty-five female Wistar rats were randomly divided into blank control group (group A), model group (group B), pre-moxibustion group (group C), instant moxibustion group (group D) and pre-instant moxibustion group (group E), with 9 rats in each group. Cold-damp coagulation and stagnation type dysmenorrhea models were established. In group C, mild moxibustion on “Shénquè (神阙 CV 8) ” and “Guānyuán (关元 CV 4)” was carried out from the time after modeling on the 8th day for 3 consecutive days. In group D, mild moxibustion was given as the same methods with group C after injection with oxytocin on the 11th day. In group E, mild moxibustion was given as the same methods from the time after modeling on the 8th day to that after injection with oxytocin on the 11th day for 4 consecutive days. The writhing behavior and the changes in levels of ET-1 and NO in uterine tissues of rats with dysmenorrhea in each group were observed. ResultsComparison of the latent period: compared with (4.38 ± 1.06) min in group B, the latent period of rats in group C (9.67 ± 1.32) min, group D (11.78 ± 1.30) min and group E (15.00 ± 1.22) min obviously prolonged (all p < 0.01). Compared with group C, the latent period of group E obviously prolonged (p < 0.01). Compared with group D, the latent period of group E obviously prolonged (p < 0.01).Comparison of the writhing times: compared with (4.38 ± 1.06) in group B, the writhing times of rats in group C (9.67 ± 1.32), group D (11.78 ± 1.30) and group E (15.00 ± 1.22) reduced (all p < 0.01). Compared with group C, the writhing times of rats in group D and group E reduced (both p < 0.01). Compared with group D, the writhing times in group E reduced (p < 0.05). Comparison of the total writhing score: compared with (4.38 ± 1.06) in group B, the total writhing score of rats in group C (9.67 ± 1.32), group D (11.78 ± 1.30) and group E (15.00 ± 1.22) decreased (all p < 0.01). Compared with group C, the total writhing score of rats in group D and group E decreased (both p < 0.01). Compared with group D, the total writhing score of rats in group E decreased (p < 0.05). Comparison of ET-1 level: compared with (4.80 ± 0.47) in group A, the ET-1 level in uterine tissues of rats in group B (7.57 ± 0.69) significantly increased (P < 0.01). Compared with group B, the ET-1 level in uterine tissues of rats in group C (6.20 ± 0.50), group D (5.67 ± 0.29) and group E (5.16 ± 0.33) obviously decreased (all p < 0.01). Compared with group C, the ET-1 level in uterine tissues of rats in group D and group E obviously decreased (p < 0.05, p < 0.01). Compared with group D, the ET-1 level in uterine tissues of rats in group Eobviously decreased (p < 0.05). Comparison of NO level: compared with (6.63 ± 1.83) in group A, the NO level in uterine tissues of rats in group B (1.62 ± 0.58) significantly decreased (p < 0.01). Compared with group B, the NO level in uterine tissues of rats in group C (3.60 ± 0.59), group D (4.77 ± 0.67) and group E (5.99 ± 0.63) obviously increased (all p < 0.01). Compared with group C, the NO level in uterine tissues of rats in group Dand group E obviously increased (p < 0.05, p < 0.01). Compared with group D, the NO level in uterine tissues of rats in group E obviously increased (p < 0.01). ConclusionThe analgesic effect of mild moxibustion at different intervention times on cold-damp coagulation and stagnation type dysmenorrhea rats was different, which was the most significant in pre-instant moxibustion group. One of the mechanisms of action may be related with the adjustment of abnormal levels of ET-1 and NO.

Brief hypoxic cycles improve uterine contractile function after prolonged hypoxia in term-pregnant but not in nonpregnant rats in vitro

During labour, the uterus itself is vulnerable to hypoxia/ischemia that can occur with each strong contraction and this may ultimately cause dysfunctional labour in some women. Periods of Intermittent re-oxygenations are beneficial to tissues subjected to hypoxia to wash out metabolic by-products that have been accumulated during hypoxic stresses which may affect the tissue viability. We proposed that short intermittent hypoxic episodes may protect the uterus from subsequent sustained long hypoxia. To investigate this, two sets of experiments were performed on term-pregnant and nonpregnant rat uterine tissues. In one set of experiment the uterus was subjected to sustained long hypoxia for 40 min and then allowed to recover in 100% O2. In the other set of experiment the uterus was subjected to 3 cycles of 2 min hypoxia each separated by 20 min reoxygenation and followed by a sustained long hypoxia for 40 min and then allowed to recover. We found that challenging the uterine tissues with intermittent short hypoxic episodes improved the uterine contractility significantly after the sustained long hypoxia in term-pregnant but not in non-pregnant tissues. These results suggest that a mechanism of uterine tolerance (preconditioning) is confined to uterine tissues very close to labour and it is a protective phenomenon to improve the uterine activity despite the long-lasting paradoxical metabolic challenges that occur during the repeated strong labour contractions.

Application of a perfusion bioreactor to promote decellularization and recellularization for rat uterine tissue

Application of a perfusion bioreactor to promote decellularization and recellularization for rat uterine tissue

Vitex agnus-castus dry extract BNO 1095 (Cyclodynon®) inhibits uterine hyper-contractions and inflammation in experimental models for primary dysmenorrhea

Background. For many women, the monthly suffering induced by menstrual “cramps” is severe enough to profoundly disrupt their quality of life. In the case of primary dysmenorrhea, a condition related to premenstrual syndrome (PMS), intense uterine contractions are thought to trigger moderate to intense pain despite the absence of an underlying infection or other medically-identifiable disease states. The associated uterine hyper-contractility is reminiscent of labor, and associated pain is likely to be mediated by the release of prostaglandins, leukotrienes and the infiltration of leukocytes that normally accompany the breakdown of the endometrial lining.Standardized extracts of Vitex agnus-castus berries (VAC extracts of chaste tree, or chaste berries) are clinically effective in treating the symptoms of PMS, yet the mechanisms of how the chemically complex mixture acts are largely unknown.Methods. Using an in vivo dysmenorrhea model rats were treated with 10 mg/kg estradiol-benzoate i.p. once daily for 12 days and with 2.1, 10.3 or 20.7 mg/kg VAC dry extract p.o. once daily for 7 days prior to induction of convulsions. Uterine contractions where induced with 2 IU/kg oxytocin i.p., followed by monitoring of abdominal convulsions and signs of pain on the last day of the experiment. Moreover, in vitro methods were applied that are described in the methods section.Results. Here, we show that the VAC herbal dry extract BNO 1095 (commercially available as Cyclodynon®) targets the uterine myometrial tissue and inflammatory signaling molecules of associated migratory/inflammatory cells. Specifically, BNO 1095 dose-dependently inhibited oxytocin-induced uterine contractions in a rat dysmenorrhea model in vivo and drug-induced contractions in isolated human and rat uterine tissue in vitro. Furthermore, BNO 1095 showed a promising anti-inflammatory capacity by potently inhibiting 5-lipoxygenase activity and leukotriene production and by reducing the production of reactive oxygen species and inflammatory cytokines in vitro.Conclusion. These results provide evidence that BNO 1095 effectively treats menstruation-related complaints including primary dysmenorrhea.

Possible effects of melatonin against rat uterus exposure to bisphenol A during neonatal period.

The aim of this study was to investigate the possible effects of melatonin on rat uterine tissue against exposure with bisphenol A (BPA) in the neonatal period. Twenty-four female rats were divided into four groups, (n=6) per group. Group I was used as a control (sesame oil + ethanol), group II was injected daily with (100mg/kg) BPA by subcutaneously (sc) daily postnatal days (PND 0-10), group III was injected daily with (10mg/kg) melatonin by sc for 10days (PND 20-30), and group IV was injected daily with (100mg/kg) BPA (PND 0-10) and (10mg/kg) melatonin (PND 20-30). All rats were sacrificed in the same day of metestrus cycle, approximately PND 70. Histological analyses, immunostaining of B cell lymphoma 2 (Bcl-2), and cytochrome c and TUNEL assays were performed. According to our results, neonatal exposure to BPA accelerates onset of puberty, causes degenerative and morphometric changes on rat uterus, and increases apoptotic reaction rates. The immunoreactivity of Bcl-2 was decreased after BPA administration. In addition, immunoreactivity of Bcl-2 showed an increase after melatonin treatment. However, cytochrome c immunoreactivity was decreased after melatonin administration. Our results suggest that melatonin may have positive effects against BPA-induced degenerative changes on rat uterus.

Effects of moxibustion at different times on prostaglandin and vasopressin levels in uterine tissues of rats with dysmenorrhea due to cold-dampness retention

To observe the effects of moxibustion at different times on prostaglandin E2 (PGE2), prostaglandin F2α (PGF2α) and arginine vasopressin (AVP), in the uterine tissues of rats with dysmenorrhea due to cold-dampness retention, and to explore the differences and possible mechanisms of moxibustion at different times in easing pain in dysmenorrhea due to cold-dampness retention. Forty-three female Wistar rats were randomly divided into a blank control group (n=7), a model group (n=9), a pre-moxibustion group (n=9), an immediate-moxibustion group (n=9) and a pre-moxibustion plus immediatemoxibustion group (n=9). Rat models of primary dysmenorrhea due to cold-dampness retention were established using (0±1) °C ice water immersion method combined with injection of estradiol benzoate for 10 d, followed by injection of oxytocin on the 11th day. Rats in the 3 intervention groups received moxibustion to Shenque (CV 8) and Guanyuan (CV 4), 10 min for each acupoint, once a day. Rats in pre-moxibustion group were given mild moxibustion, beginning on the 8th day during modeling, for 3 continuous days; rats in immediate-moxibustion group were given one time mild moxibustion, immediately after injection of oxytocin on the 11th day during modeling; rats in pre-moxibustion plus immediate-moxibustion group were given mild moxibustion, beginning on the 8th day during modeling till immediately after injection of oxytocin on the 11th day during modeling, for 4 continuous days. The level of PGF2α in the rat uterine tissues was measured by enzyme-linked immunosorbent assay (ELISA), and the levels of PGE2 and AVP in rat uterine tissues were measured by radioimmunoassay. Compared with the blank control group, the levels of PGF2α and AVP, the PGF2α/PGE2 ratio in the model group were significantly increased (P<0.01), and the PGE2 level was significantly decreased (P<0.01) in the rat uterine tissues in the model group. Compared with the model group, the writhing latency was significantly prolonged, the writhing number and the total writhing score were all decreased in the pre-moxibustion group, the immediate-moxibustion group and the pre-moxibustion plus immediate-moxibustion group (all P<0.01); the levels of PGF2α and AVP, and the PGF2α/PGE2 ratio were all significantly decreased (P<0.05, P<0.01), and the PGE2 level was significantly increased (P<0.01) in the rat uterine tissues of the 3 treatment groups. Compared with the pre-moxibustion group, the writhing number and the total writhing score were all decreased in the immediate-moxibustion group and the pre-moxibustion plus immediate-moxibustion group (all P<0.01), the writhing latency was significantly prolonged in the pre-moxibustion plus immediate-moxibustion group(P<0.01); the levels of PGF2α and PGF2α/PGE2 ratio were significantly decreased (P<0.05, P<0.01), and the PGE2 level was significantly increased (P<0.01) in rat uterine tissues in the immediate-moxibustion group and the pre-moxibustion plus immediate-moxibustion group. Compared with the immediate-moxibustion group, the writhing latency was significantly prolonged and the writhing number was decreased (all P<0.05), and the total writhing score was decreased (P<0.01) in the pre-moxibustion plus immediatemoxibustion group; the PGF2α level and the PGF2α/PGE2 ratio were significantly decreased (P<0.01), and the PGE2 level was significantly increased (P<0.01) in rat uterine tissues in the pre-moxibustion plus immediate-moxibustion group. Moxibustion at different times all can produce obvious analgesic effects on dysmenorrhea due to cold-dampness retention in rats, and pre-moxibustion plus immediate-moxibustion ranks the top. The mechanism of this analgesic effect may be via the regulation of abnormal PGF2α, PGE2 and AVP levels, to effectively inhibit the spastic contraction of uterine smooth muscle in dysmenorrhea rat, thereby improving the ischemia and hypoxia in uterus.

Effect of instant moxibustion on the levels of prostaglandin and arginine vasopressin in the uterine tissues of dismenorrhea rats with cold-damp congealing and stagnation type

ObjectiveTo observe the effect of instant moxibustion on the levels of prostaglandin E2 (PGE2), prostaglandin F2α (PGE2α) and arginine vasopressin (AVP) in the uterine tissues of dismenorrhea rats with cold-damp congealing and stagnation type and to explore its possible mechanism. MethodsFemale Wistar rats were randomly divided into blank group, model group, herble medicine group, pre-moxibustion group and instant moxibustion group, with 9 rats in each group. Cold-damp congealing and stagnation type primary dismenorrhea models were established by adopting (0±1)°C ice water-extraction method combined with estradiol benzoate injection method. After modeling on the 8th day, in herble medicine group, Tongjingbao granules was given to the rats by intragastric administration. In pre-moxibustion group, mild moxibustion was carried out at “Shénquè” (▪ CV 8) and “Guānyuán” (▪ CV 4) of the rats for 10 min at each acupoint. In instant moxibustion group, moxibustion as that in pre-moxibustion group was conducted for once after injection with oxytocin on the 11th day. ELISA was adopted to detect the levels of PGE2 and PGE2α in the uterine tissues of rats, and radioimmunoassay was used for detection of AVP level in the uterine tissues of rats. ResultsCompared with the model group, the latent period of rats in herbal medicine group, pre-moxibustion group and instant moxibustion group obviously prolonged, the number of times of torsion reduced, and the total score of torsion decreased (P<0.01); compared with herbal medicine group, the latent period of rats in instant moxibustion group obviously prolonged, and the total score of torsion decreased (P<0.05 or P<0.01); compared with pre-moxibustion group, the number of times of torsion of rats in instant moxibustion group reduced, and the total score of torsion decreased (P<0.01). Compared with blank group, the levels of PGE2α and AVP and the ratio of PGE2α and PGE2 in the uterine tissues of rats in model group significantly increased (P<0.01), and the PGE2 level significantly reduced (P<0.01); compared with model group, the PGE2α level and the ratio of PGE2α and PGE2 in the uterine tissues of rats in herble medicine group, pre-moxibustion group and instant moxibustion group obviously reduced (P<0.05 or P<0.01), the PGE2 level obviously increased (P<0.01), and the AVP level in the uterine tissues of rats in pre-moxibustion group and instant moxibustion group obviously reduced (P<0.05 or P<0.01); compared with herbal medicine group, the levels of PGE2α and AVP and the ratio of PGE2α and PGE2 in the uterine tissues of rats in instant moxibustion group significantly reduced (P<0.05 or P<0.01); compared with pre-moxibustion group, the PGE2α level and the ratio of PGE2α and PGE2 in the uterine tissues of rats in instant moxibustion group obviously reduced (P<0.05), and the PGE2 level obviously increased (P<0.01). ConclusionBoth pre-moxibustion and instant moxibustion can obviously inhibit spasmodic uterine smooth muscle contraction of rats with dismenorrhea, regulate imbalanced levels of PGE2α and PGE2, reduce the AVP level, so as to improve the uterine hypoxia-ischemia, and play a role in alleviating pain. The efficacy of instant moxibustion was superior to that of pre-moxibustion.

Selective Estrogen Receptor Modulator (SERM)-like Activities of Diarylheptanoid, a Phytoestrogen from Curcuma comosa, in Breast Cancer Cells, Pre-osteoblast Cells, and Rat Uterine Tissues.

Diarylheptanoids from Curcuma comosa, of the Zingiberaceae family, exhibit diverse estrogenic activities. In this study we investigated the estrogenic activity of a major hydroxyl diarylheptanoid, 7-(3,4 -dihydroxyphenyl)-5-hydroxy-1-phenyl-(1E)-1-heptene (compound 092) isolated from C. comosa. The compound elicited different transcriptional activities of estrogen agonist at low concentrations (0.1-1 μM) and antagonist at high concentrations (10-50 μM) using luciferase reporter gene assay in HEK-293T cells. In human breast cancer (MCF-7) cells, compound 092 showed an anti-estrogenic activity by down-regulating ERα-signaling and suppressing estrogen-responsive genes, whereas it attenuated the uterotrophic effect of estrogen in immature ovariectomized rats. Of note, compound 092 promoted mouse pre-osteoblastic (MC3T3-E1) cell differentiation and the related bone markers, indicating its positive osteogenic effect. Our findings highlight a new, nonsteroidal, estrogen agonist/antagonist of catechol diarylheptanoid from C. comosa, which is scientific evidence supporting its potential as a dietary supplement to prevent bone loss with low risk of breast and uterine cancers in postmenopausal women.

Vitex agnus-castus dry extract BNO 1095 (Agnucaston\xae) inhibits uterine hyper-contractions and inflammation in experimental models for primary dysmenorrhea

BackgroundFor many women, the monthly suffering induced by menstrual “cramps” is severe enough to profoundly disrupt their quality of life. In the case of primary dysmenorrhea, a condition related to premenstrual syndrome (PMS), intense uterine contractions are thought to trigger moderate to intense pain despite the absence of an underlying infection or other medically-identifiable disease states. The associated uterine hyper-contractility is reminiscent of labor, and associated pain is likely to be mediated by the release of prostaglandins, leukotrienes and the infiltration of leukocytes that normally accompany the breakdown of the endometrial lining.Standardized extracts of Vitex agnus-castus berries (VAC extracts of chaste tree, or chaste berries) are clinically effective in treating the symptoms of PMS, yet the mechanisms of how the chemically complex mixture acts are largely unknown.MethodsUsing an in vivo dysmenorrhea model rats were treated with 10 mg/kg estradiol-benzoate i.p. once daily for 12 days and with 2.1, 10.3 or 20.7 mg/kg VAC dry extract p.o. once daily for 7 days prior to induction of convulsions. Uterine contractions where induced with 2 IU/kg oxytocin i.p., followed by monitoring of abdominal convulsions and signs of pain on the last day of the experiment. Moreover, in vitro methods were applied that are described in the methods section.ResultsHere, we show that the VAC herbal dry extract BNO 1095 (commercially available as Agnucaston®) targets the uterine myometrial tissue and inflammatory signaling molecules of associated migratory/inflammatory cells. Specifically, BNO 1095 dose-dependently inhibited oxytocin-induced uterine contractions in a rat dysmenorrhea model in vivo and drug-induced contractions in isolated human and rat uterine tissue in vitro. Furthermore, BNO 1095 showed a promising anti-inflammatory capacity by potently inhibiting 5-lipoxygenase activity and leukotriene production and by reducing the production of reactive oxygen species and inflammatory cytokines in vitro.ConclusionThese results provide evidence that BNO 1095 effectively treats menstruation-related complaints including primary dysmenorrhea.

Selenium Plays a Protective Role in Staphylococcus aureus-Induced Endometritis in the Uterine Tissue of Rats.

The essential trace element selenium (Se) modulates the functions of many regulatory proteins in signal transduction, conferring benefits in inflammatory diseases. Endometritis is a reproductive obstacle disease both in humans and animals. Staphylococcus aureus is the major pathogen that causes endometritis. The present study analyzes the protection and mechanism of Se-methylselenocysteine (MSC) and methylseleninic acid (MSA) on S. aureus-induced endometritis. An atomic fluorescence spectrophotometry study showed that the uterine Se content increased with the addition of MSC and MSA. Histopathology observation and TUNEL detection showed that Se supplementation displayed a greater defense against uterine inflammatory damage. The quantitative PCR (qPCR) and ELISA analyses showed that the expressions of tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6) increased with S. aureus infection and decreased with the addition of MSC and MSA. The Toll-like receptor 2 (TLR2) expression showed the same status as the inflammatory cytokines. The Western blot results showed that the increased phosphorylation of IκBα and NF-κB p65 was also reduced by the addition of MSC and MSA. The qPCR and Western blot results also showed that the transcription expressions and the protein dissociation of caspase-9, caspase-3, caspase-7, caspase-6, and poly(ADP-ribose) polymerase (PARP), which were increased by S. aureus infection, were inhibited by Se supplementation. All of the results displayed that the protection conferred by MSC was stronger than MSA. The present study indicated the Se supplementation might be a potential prevention and control measure for S. aureus-induced endometritis.

Morphological Changes in Rat Uterine Tissues and Possibility of Spontaneous Labor as a Result of Injection of Multipotent Mesenchymal Stromal Cells against the Background of Hydrometra.

The possibility of pregnancy and labor was evaluated and tissue changes after injection of autologous bone marrow multipotent mesenchymal stromal cells with transfected GFP gene were studied in rats with experimental hydrometra. Injection of stromal cells to the uterine cicatrix increased the number of vessels (vascular walls or their elements) formed de novo with participation of injected cells. The animals produced progeny 2 estrous cycles earlier, the percentage of "puerperal" rats in this group was higher, their progeny was more numerous and they had the maximum numbers of little rats. The maternal mortality was lower after injection of stromal cells. Injection of stromal cells led to development of a trend to more rapid reparative processes in the uterus in animals with cicatricial stenosis of its lumen.