Brain/metabolism* ; Oxytocin ; Light

This study aimed to investigate the therapeutic effect of Leonuri Herba aqueous decoction on primary dysmenorrhea(PD) and explore the underlying mechanism in conjunction with untargeted metabolomics. Forty adult female rats were randomly divi-ded into a normal group, a model control group, ibuprofen(0.12 g·kg~(-1)) group, and high-and low-dose Leonuri Herba aqueous decoction(5 and 2.5 g·kg~(-1)) groups, with eight rats in each group. The PD rat model was prepared using intramuscular injection of estradiol benzoate combined with intraperitoneal injection of pitocin. Drugs were administered by gavage from the 4th day of modeling for 7 d. After the last administration, pitocin was injected intraperitoneally, and the writhing latency and writhing times within 30 min were recorded. The uterine and ovarian coefficients were determined. Estradiol(E_2), progesterone(Prog), oxytocin(OT), cyclooxyge-nase 2(COX-2), prostaglandin E_2(PGE_2), prostaglandin F_(2α)(PGF_(2α)), and Ca~(2+) levels in uterine tissues were measured by ELISA and biochemical kits. Morphological changes in uterine and ovarian tissues were observed by hematoxylin-eosin(HE) staining. The protein expression of oxytocin receptor(OTR), prostaglandin E_2 receptor 3(EP3), and estrogen receptor alpha(ERα) in uterine tissues was detected by immunohistochemistry. The mRNA expression of OTR, PGE_2 receptors 1-4(EP1, EP2, EP3, and EP4), and PGF_(2α) receptor(FP) in uterine tissues was detected by quantitative real-time PCR. Untargeted metabolomics analysis was performed by ultra-high-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(LC-QTOF-MS) technology to screen potential biomarkers and enrich metabolic pathways. The results showed that Leonuri Herba was able to significantly reduce the writhing times in PD rats(P<0.05 or P<0.01), significantly reduce the uterine and ovarian coefficients(P<0.01), and improve their histomorphology. After treatment with Leonuri Herba, PGE_2 content was significantly increased(P<0.05), COX-2, PGF_(2α) and Ca~(2+) content, and PGF_(2α)/PGE_2 was significantly decreased(P<0.05 or P<0.01), and OT content was decreased, while E_2 and Prog content tended to further increase in uterine tissues of PD rats. Correspondingly, OTR and EP3 protein expression was significantly downregulated(P<0.05 or P<0.01) and ERα protein expression was upregulated(P<0.05) in uterine tissues. The mRNA expression of FP and EP4 in uterine tissues was significantly downregulated(P<0.01), and the mRNA expression of EP1, EP3, and OTR showed a decreasing trend. The untargeted metabolomics results showed that 10 differential metabolites were restored in the plasma of PD rats after Leonuri Herba treatment. The results indicate that Leonuri Herba is effective in the prevention and treatment of PD, and the underlying mechanism may be attributed to the regulation of PGs synthesis and corresponding receptor binding.
Humans ; Rats ; Female ; Animals ; Estrogen Receptor alpha ; Oxytocin ; Dysmenorrhea/metabolism* ; Cyclooxygenase 2 ; Dinoprostone ; RNA, Messenger/metabolism* ; Dinoprost

To study preliminarily the effect of Jiawei Bazhen decoction combined with oxytocin in promoting cervical ripening of full-term pregnancy women who were in the deficiency of qi and blood type through the syndrome differentiation of traditional Chinese medicine (TCM). 180 patients that met the inclusion criteria of the study were randomly divided into three groups: the control group(oxytocin group), the treatment group (Jiawei Bazhen decoction combined with oxytocin group), the blank control group (expected and observation group). Cervical maturity score (Bishop score), vaginal and cervical secretions fetal fibronectin (FFN), the result of induced labor, the result of mother and baby were observed in each group before and after treatment. The result comes out that the cervical Bishop score of pregnant women for treatment group were significantly higher than the control group and blank control group after treatment (P < 0.05). The FFN of pregnant women for the treatment group were significantly different from the control group and blank control group after treatment (P < 0.05). The pregnancy outcome of the three groups: the labor rate and rate of vaginal delivery of the treatment group were higher than the other two groups, and the difference was statistically significant (P < 0.05). The cesarean section rate of the treatment group was significantly lower than the other two groups, the difference was also statistically significant (P < 0.05). The three groups did not appear the phenomenon of neonatal asphyxia. Jiawei Bazhen decoction combined with oxytocin is effective in producing cervical ripening and induce labor. It is convenient, safe and reliable, for it is no obvious adverse effects on mother and fetus, but effective in reducing the rate of cesarean section, and playing a positive role in promoting natural delivery.
Adult ; Cervical Ripening ; drug effects ; metabolism ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Fibronectins ; secretion ; Humans ; Labor, Induced ; Oxytocin ; administration & dosage ; Pregnancy ; Pregnancy Complications ; drug therapy ; metabolism ; physiopathology ; Pregnancy Outcome ; Qi ; Vagina ; drug effects ; secretion ; Young Adult

OBJECTIVETo analyze the level of the oxytocin (OT) and the expression of oxytocin receptor (OTR) in males with idiopathic infertility.

METHODSSixty-five infertile males aged 20 -45 years were divided according to their semen parameters into an idiopathic oligozoospermia group (OG, n = 20), an idiopathic asthenozoospermia group (AG, n = 25), and an idiopathic oligoasthenozoospermia group (OAG, n = 20). Another twenty 20-45 years old healthy male volunteers with a natural childbearing history were included in the control group (CG). All the subjects were detected for the contents of luteinizing hormone (LH), follicle stimulating hormone (FSH) , testosterone (T) and OT, and analyzed for the expression of OTR by sequencing the functional region of the OTR promoter (OTRP), OTR-mRNA, and OTR-COOH terminus. The gene sequences were compared using DNASTAR-MegAlign, Western blot values changed into enumeration data, and all the data analyzed by one-way ANOVA and Dunnette's multiple range t-test.

RESULTSA significantly lower content of OT was observed in CG ( [79.30 +/- 3.83] pg/ml) than in OG ([118.53 +/- 7.69] pg/ml, AG ([108.81 +/- 5.66] pg/ml) and OAG ([103.71 +/- 4.54] pg/ml) (F(0.05/2[2,82]) = 8.29, P < 0.01). The content of LH was significantly lower in AG ([4.26 +/- 0.31] IU/L) and OAG ([4.55 +/- 0.40] IU/L) than in OG ([6.77 +/- 0.57] IU/L) and CG ([7.19 +/- 0.50] IU/L) (F(0.05/2 [2,82]) = 11.64, P < 0.01), and so was the content of FSH in AG ( [5.02 + 0.39] IU/L) than in CG ([8.91 +/- 0.91] IU/L), OG ([11.86 +/- 1.76] IU/L) and OAG ([8.82 +/- 1.03] IU/L) (F(0.05/[2,82]) = 7.22, P < 0.01). There were no significant differences in the T content among the four groups (F(0.05/2[2,82] = 0.42, P = 0.739). No evident gene mutation was found in OTRP and OTR-mRNA gene sequencing. Human OTRs in the lymphocytes were monomers and oligomers, mostly tetramers and hexamers. There were obviously more monomers in AG (0.41 +/- 0.03) and OAG (0.13 +/- 0.01) than in OG (0.05 +/- 0.004) and CG (0.05 +/- 0.003) (F(0.05/2[2,82]) = 115.50, P < 0.01), while the number of oligomers was markedly decreased in 20% of the cases in AG.

CONCLUSIONSignificant differences in the content of OT and expression of OTR between fertile and infertile men suggested an association of OT with male infertility. The decreased expression of OTR oligomers and increased expression of monomers may be related to idiopathic asthenozoospermia, which has provided a new insight into the pathogenesis and treatment of male infertility.

Adult ; Case-Control Studies ; Humans ; Infertility, Male ; metabolism ; pathology ; Male ; Middle Aged ; Neuropeptides ; metabolism ; Oxytocin ; metabolism ; Receptors, Oxytocin ; metabolism ; Young Adult

OBJECTIVETo probe the mechanism of acupuncture in treatment of dysmenorrhea.

METHODSAdult mice with no pregnancy were randomly divided into a normal group, a model group, an acupuncture group and a medication group. The model group, the acupuncture group and the medication group were modeled by Diethylstilbestrol and Ocytocin. For the acupuncture group, at the 7th day of modeling, acupuncture was given at "Sanyinjiao" (SP 6), "Diji" (SP 8), once a day, for 5 days; and at the 7th day of modeling, Yimucao Gao 0.6 mg/g was given intragastrically to the medication group for 5 days. The stretching latent period and the number of stretching within 30 min were observed, and mRNA levels of ocytocin receptor (OctR) and vasopressin receptor (VasR) in the uterus tissue were detected with RT-PCR method.

RESULTSCompared with the model group, the stretching latent period extended (P < 0.05) and the number of stretching within 30 min significantly decreased (P < 0.05); and there were significant differences in the mRNA levels of ocytocin receptor and vasopressin receptor in the uterus tissue in the model group as compared with those in other 3 groups (P < 0.05, P < 0.01).

CONCLUSIONAcupuncture can improve the dysmenorrheal symptom to a certain extent, and the mechanism is possibly related to regulative effects of acupuncture on hormones-mediating receptors in mice.

Acupuncture Therapy ; Animals ; Dysmenorrhea ; metabolism ; therapy ; Female ; Mice ; RNA, Messenger ; analysis ; Receptors, Oxytocin ; genetics ; Receptors, Vasopressin ; genetics

OBJECTIVETo observe the influence of silencing Connexin43 (Cx43) expression of partial acupoints on acupuncture effect, so as to probe into the mechanism of acupuncture treatment for primary dysmenorrhea.

METHODSThe primary dysmenorrheal rat model made by oxytocin and RNA interference (RNAi) technology was used to silence the expression of Cx43 in acupoints. Fifty SD female rats were divided into five groups, a normal group (N), a model group (M), an acupuncture group (A), an acupuncture plus interference group (A+I), an acupuncture plus interference control group (A+IC). RT-PCR method was used to observe the oxytocin receptor (OTR) and vasopressin receptor (VPR) mRNA expressions in the uterus in each group. Plasma prostaglandin E2 (PGE2) and PGF2alpha levels were detected by radioimmunoassay and ELISA, respectively.

RESULTS(1) The times of writhing body (9.43 +/- 3.87 and 10.28 +/- 4.23) were significantly lower and the latency period of writhing body (12.43 +/- 3.46, 11.00 +/- 3.65) were longer in the group A and the group A+IC as compared with (15.43 +/- 5.13, 17.00 +/- 3.87) and (7.57 +/- 1.99, 8.43 +/- 2.57) in the group M and group A+I (P < 0.05), respectively. (2) The levels of Cx43 mR NA level and protein expression of acupoint in the group A+I were significantly lower than those of the group N (P < 0.05). (3) OTR and VPR mRNA in the uterus in the group A and the group A+IC were significantly lower than those in the group M and the group A+I (P < 0.05), with no significant difference between the group M and the group A+I (P > 0.05). (4) As compared with the group M, PGE2 level increased and PGF2alpha level decreased in the group A and the group A+IC (P < 0.05).

CONCLUSIONSilencing Cx43 expression of partial acupoint can inhibit effectively the effect of acupuncture through decreasing OTR and VPR in endometrium of the dysmenorrheal rat and adjusting the prostaglandins (PGs) synthesis system, which possibly is one of the mechanisms of acupuncture for treatment of primary dysmenorrhea.

Acupuncture Points ; Acupuncture Therapy ; Animals ; Connexin 43 ; genetics ; metabolism ; Dinoprost ; blood ; Dinoprostone ; blood ; Dysmenorrhea ; genetics ; metabolism ; therapy ; Female ; Gene Expression ; Humans ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Oxytocin ; genetics ; metabolism ; Receptors, Vasopressin ; genetics ; metabolism ; Uterus ; metabolism

Organotypic slice cultures have been developed as an alternative to acute brain slices because the neuronal viability and synaptic connectivity in these cultures can be preserved well for a prolonged period of time. This study evaluated a stationary organotypic slice culture developed for the hypothalamic paraventricular nucleus (PVN) of rat. The results showed that the slice cultures maintain the typical shape of the nucleus, the immunocytochemical signals for oxytocin, vasopressin, and corticotropin-releasing hormone, and the electrophysiological properties of PVN neurons for up to 3 weeks in vitro. The PVN neurons in the culture expressed the green fluorescent protein gene that had been delivered by the adenoviral vectors. The results indicate that the cultured slices preserve the properties of the PVN neurons, and can be used in longterm studies on these neurons in vitro.
Adenoviridae ; Animals ; Cell Culture Techniques/*methods ; Corticotropin-Releasing Hormone/metabolism ; Electrophysiology ; Genetic Vectors ; Green Fluorescent Proteins/metabolism ; Immunohistochemistry ; Neurons/*cytology/metabolism ; Oxazines ; Oxytocin/metabolism ; Paraventricular Hypothalamic Nucleus/*anatomy & histology/cytology/metabolism ; Rats ; Vasopressins/metabolism

Chromosome Aberrations ; Diagnosis, Differential ; Endometrial Neoplasms ; genetics ; metabolism ; pathology ; Endometrial Stromal Tumors ; genetics ; metabolism ; pathology ; Female ; Histone Deacetylases ; metabolism ; Humans ; Immunohistochemistry ; Neprilysin ; metabolism ; Receptors, Oxytocin ; metabolism ; Repressor Proteins ; metabolism ; Sarcoma, Endometrial Stromal ; genetics ; metabolism ; pathology ; Uterine Neoplasms ; metabolism ; pathology

OBJECTIVE: Oxytocin antagonists maybe useful in inhibiting the uterine contractions of preterm labor. One such compound is TT-235. The purpose of this study was to compare the resistance of TT-235 and oxytocin to enzymatic degradation by oxytocinase in pregnant human. METHODS: Blood samples from pregnant women not in labor were incubated in vitro with known amounts of oxytocin and TT-235. Samples were collected at 0, 15, 30, 45 and 60 minute intervals for oxytocin analysis and at 0, 10, 60 and 360 minutes for TT-235 analysis. Oxytocin was analyzed by radioimmunoassay after extraction while TT-235 was analyzed by radioreceptor assay. RESULTS: In human blood, oxytocin was readily metabolized with greater than 83% disappearance over the 60 minute incubation period. In contrast, TT-235 was stable up to 360 minutes of incubation. CONCLUSION: This study suggests that: (1) blood from pregnant human does contain oxytocinase at least in vitro; and (2) TT-235 was resistant to enzymatic degradation by human blood, implying that this oxytocin antagonist may have prolonged activity in vivo in humans.
Cystinyl Aminopeptidase ; Female ; Humans ; Metabolism* ; Obstetric Labor, Premature ; Oxytocin* ; Plasma* ; Pregnancy ; Pregnant Women* ; Radioimmunoassay ; Radioligand Assay ; Uterine Contraction ; Uterus

This study was performed to determine the action mode of oxytocin antagonist. In Study 1, the duration of in vivo action of oxytocin antagonist I (AI) was examined. After infusing AI, oxytocin was given and repeated every hour for 5 hr. Uterine activities were monitored with a polygraph. Study 2 determined the effect of AI on uterine oxytocin receptor number (Rn) and binding affinity (Kd). AI treated rats were sacrificed at 0.5 and 4 hr later for receptor assay. In Study 1, the uterine contractile response to oxytocin was significantly inhibited (p>0.05) compared to controls at five min, 1 and 2 hr after injection of AI. No differences in response were detected compared to controls (p>0.05) at later hours. In Study 2, no differences (p>0.05) between the AI and control animals in either oxytocin receptor number or binding affinity was found. These data suggest that the major mode of AI action is via competitive inhibition at the uterine oxytocin receptor and not by altering receptor number or binding affinity. AI is suggested to have the potential of being a potent and specific tocolytic agent for prevention of preterm labor in human.
Animal ; Female ; Oxytocin/pharmacology ; Oxytocin/metabolism ; Oxytocin/antagonists & inhibitors* ; Rats ; Receptors, Oxytocin/metabolism ; Uterus/physiology ; Uterus/drug effects*