Animals ; Cell Hypoxia ; Glucose ; metabolism ; Mifepristone ; pharmacology ; Oxidative Stress ; PC12 Cells ; Progesterone ; pharmacology ; Rats

OBJECTIVETo observe the effect of electroacupuncture (EA) of different acupoints on abnormal electrohysterogram (EHG) in pregnant rats, so as to analyze their regularities in regulating dysfunction of the viscera.

METHODSA total of 48 Wistar pregnant rats (18-20 days) anesthetized with mixture solution of 1.5% chloralose and 25% urethane (i. p) were randomized into control (n=10), "Sanyinjiao" (SP 6, n=9), "Hegu" (LI 4, n=8), "Neiguan" (PC 6, n=0), and "SP 6 + LI 4" (n=11) groups. EHG was recorded by using a bipolar stainless steel electrode inserted in the sub-perimetrium layer of the left mid part of the uterus. The reference electrode was placed beneath the skin of the incision. Oxytocin and gesterol were given to the local uterus nearby the recording electrode to induce abnormal excitement and suppression of EHG respectively. EA (1-2 mA, 2/15 Hz) was applied to the above-mentioned acupoints separately for 20 min in the EA groups and the influences of EA on the amplitude and frequency of burst (fast) waves and slow waves of EHG were analyzed.

RESULTSCompared with the control group, EA of SP 6 + LI 4 and SP 6 groups had apparent inhibitory effects on oxytocin-induced increases of the frequency and amplitude of both fast and slow waves (P<0.05); and EA of LI 4 also had a markedly inhibitory effect on the amplitude of fast waves (P<0.05). No marked effects on both frequency and amplitude of fast waves and slow waves of EHG were found in the PC 6 group (P>0.05). In comparison with the control group, EA of SP 6+LI 4 and SP 6 could relieve or significantly relieve progesterone-induced suppression of the frequency and amplitude of both fast and slow waves (P<0.05); and the effects of SP 6 + LI 4 appeared earlier and lasted longer than those of SP 6; while EA of LI 4 and PC 6 had no obvious effect on progesterone-induced changes of the frequency and amplitude of both fast and slow waves (P>0.05).

CONCLUSIONEA different acupoints have their own relative specificity in regulating abnormal EHG.

Acupuncture Points ; Animals ; Electroacupuncture ; Female ; Myometrium ; physiology ; Oxytocin ; pharmacology ; Pregnancy ; Pregnancy, Animal ; physiology ; Progesterone ; pharmacology ; Random Allocation ; Rats ; Rats, Wistar

The acrosome reaction is a prerequisite for fertilization, and its signaling pathway has been investigated for decades. Regardless of the type of inducers present, the acrosome reaction is ultimately mediated by the elevation of cytosolic calcium. Inositol 1,4,5-trisphosphate-gated calcium channels are important components of the acrosome reaction signaling pathway and have been confirmed by several researchers. In this study, we used a novel permeabilization tool BioPORTER^® and first demonstrated its effectiveness in spermatozoa. The inositol 1,4,5-trisphosphate type-1 receptor antibody was introduced into spermatozoa by BioPORTER^® and significantly reduced the calcium influx and acrosome reaction induced by progesterone, solubilized zona pellucida, and the calcium ionophore A23187. This finding indicates that the inositol 1,4,5-trisphosphate type-1 receptor antibody is a valid inositol 1,4,5-trisphosphate receptor inhibitor and provides evidence of inositol 1,4,5-trisphosphate-gated calcium channel involvement in the acrosome reaction in human spermatozoa. Moreover, we demonstrated that the transfer of 1,4,5-trisphosphate into spermatozoa induced acrosome reactions, which provides more reliable evidence for this process. In addition, by treating the spermatozoa with inositol 1,4,5-trisphosphate/BioPORTER^® in the presence or absence of calcium in the culture medium, we showed that the opening of inositol 1,4,5-trisphosphate-gated calcium channels led to extracellular calcium influx. This particular extracellular calcium influx may be the major process of the final step of the acrosome reaction signaling pathway.
Acrosome Reaction/physiology* ; Calcimycin/pharmacology* ; Calcium/pharmacology* ; Calcium Ionophores/pharmacology* ; Drug Delivery Systems ; Humans ; Inositol 1,4,5-Trisphosphate Receptors/metabolism* ; Male ; Progesterone/pharmacology* ; Spermatozoa/metabolism* ; Zona Pellucida/metabolism*

OBJECTIVEObserving the effect of phenolic acids from Arnebia euchroma assist mifepristone in anti-early pregnancy of SD rattus norvegicus.

METHODFeed the SD rattus norvegicus with phenolic acids from A. euchroma during the 7 th to 9 th day, and then we observe the restaining rate of pregnancy. At the same time, we determine the progesterone level in blood serum in the ways of radioimmunoassay.

RESULT720 g x kg(-1) enolic aids from A. euchroma can markedly increase the restaining rate of pregnancy (P < 0.05) than that only mifepristone dose (8.0 g x kg(-1)). In addition, the number of everage still bith increase, however, to the pogesterone level in blood serum. It has little effect.

CONCLUSIONThe effect of phenolic acids from A. euchroma assist mifepristone in anti-early pregnancy of SD rattus norvegicus is clear, and it dosen't work in the ways of decreasing the pogesterone level.

Abortifacient Agents ; chemistry ; pharmacology ; Animals ; Boraginaceae ; chemistry ; Female ; Hydroxybenzoates ; chemistry ; pharmacology ; Male ; Mifepristone ; pharmacology ; Pregnancy ; drug effects ; Progesterone ; blood ; Radioimmunoassay ; Rats ; Rats, Sprague-Dawley

OBJECTIVE: This study aimed to compare 9 perfluoroalkyl sulfonic acids (PFSA) with carbon chain lengths (C4-C12) to inhibit human placental 3β-hydroxysteroid dehydrogenase 1 (3β-HSD1), aromatase, and rat 3β-HSD4 activities. METHODS: Human and rat placental 3β-HSDs activities were determined by converting pregnenolone to progesterone and progesterone secretion in JEG-3 cells was determined using HPLC/MS-MS, and human aromatase activity was determined by radioimmunoassay. RESULTS: PFSA inhibited human 3β-HSD1 structure-dependently in the order: perfluorooctanesulfonic acid (PFOS, half-maximum inhibitory concentration, IC ₅₀: 9.03 ± 4.83 μmol/L) > perfluorodecanesulfonic acid (PFDS, 42.52 ± 8.99 μmol/L) > perfluoroheptanesulfonic acid (PFHpS, 112.6 ± 29.39 μmol/L) > perfluorobutanesulfonic acid (PFBS) = perfluoropentanesulfonic acid (PFPS) = perfluorohexanesulfonic acid (PFHxS) = perfluorododecanesulfonic acid (PFDoS) (ineffective at 100 μmol/L). 6:2FTS (1H, 1H, 2H, 2H-perfluorooctanesulfonic acid) and 8:2FTS (1H, 1H, 2H, 2H-perfluorodecanesulfonic acid) did not inhibit human 3β-HSD1. PFOS and PFHpS are mixed inhibitors, whereas PFDS is a competitive inhibitor. Moreover, 1-10 μmol/L PFOS and PFDS significantly reduced progesterone biosynthesis in JEG-3 cells. Docking analysis revealed that PFSA binds to the steroid-binding site of human 3β-HSD1 in a carbon chain length-dependent manner. All 100 μmol/L PFSA solutions did not affect rat 3β-HSD4 and human placental aromatase activity. CONCLUSION Carbon chain length determines inhibitory potency of PFSA on human placental 3β-HSD1 in a V-shaped transition at PFOS (C8), with inhibitory potency of PFOS > PFDS > PFHpS > PFBS = PFPS = PFHxS = PFDoS = 6:2FTS = 8:2FTS.
Humans ; Pregnancy ; Female ; Rats ; Animals ; Placenta ; Progesterone/pharmacology* ; Aromatase/pharmacology* ; Cell Line, Tumor ; Fluorocarbons ; Alkanesulfonic Acids ; Structure-Activity Relationship ; Hydroxysteroid Dehydrogenases/pharmacology*

Action Potentials ; drug effects ; physiology ; Animals ; Female ; Guinea Pigs ; Heart Ventricles ; drug effects ; Papillary Muscles ; drug effects ; physiology ; Progesterone ; pharmacology

OBJECTIVETo investigate the effects of progesterone on the growth and migration of breast cancer cells.

METHODSMCF-7 and T-47D cells were cultured in DMEM and stimulated with 100 nmol/L progesterone for 48 h, and the cell proliferation was evaluated by MTT assay, cell migration by wound-healing assay and E-catherin expression by Western blotting.

RESULTSProgesterone stimulated the cell proliferation and migration and down-regulated the expression of E-catherin in both MCF-7 and T-47D cells.

CONCLUSIONSProgesterone stimulates the cell proliferation and migration of cultured breast cancer cells, suggesting the clinical significance of anti-progesterone therapy in breast cancer.

Breast Neoplasms ; pathology ; Cadherins ; metabolism ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Female ; Humans ; Progesterone ; pharmacology ; Tumor Cells, Cultured

Animals ; In Vitro Techniques ; Muscle Contraction ; drug effects ; Muscle, Smooth ; drug effects ; physiology ; Progesterone ; pharmacology ; Rabbits ; Trachea ; drug effects

OBJECTIVETo observe the relative activity of sperm mitochondria and the proportion of ROS-positive sperm before and after capacitation and progesterone (Pg)-induced hyperactivation, and investigate the functional characteristics of sperm mitochondria.

METHODSWe collected 20 samples of normal human spermatozoa that met the criteria of WHO Laboratory Manual for the Examination and Processing of Human Semen (5th ed) and cultured them with the swim-up method in a CO2 incubator at 37 degrees C for 1 hour. We divided the sperm into a pre-capacitation and a capacitated group, and further incubated the capacitated sperm in an upright tube with (Pg-induced group) or without (control group) slow-releasing Pg at 37 degrees C for another hour. Then we determined the relative activity of mitochondria and the percentage of ROS-positive cells in the sperm samples using JC-1 and DCF staining.

RESULTSThe relative activities of mitochondria were significantly increased in the capacitated, control and Pg-induced groups (6.23, 14.36 and 12.33) as compared with the pre-capacitation group (1.42) (P < 0.05), while the percentages of balanced mitochondria (mitochondria with equal amount of high and low electric potentials) remarkably reduced (4.27%, 5.03% and 8.57% vs 21.64%, P < 0.05). The percentages of ROS-positive sperm in the pre-capacitation, capacitated, control and Pg-induced groups were 2.89%, 0.70%, 4.25% and 1.90%, respectively, significantly lower in the capacitated than in the pre-capacitation group (P < 0.01), but dramatically increased in the control group after another hour of swim-up incubation and markedly higher than in the Pg-induced group (P < 0.01).

CONCLUSIONProgesterone induction can hyperactive human sperm motility, inhibit the relative activity of mitochondria, keep mitochondria potential at a more balanced level, and reduce the production of ROS, which may help to raise the rate of in vitro fertilization and improve the quality of embryos.

Adult ; Humans ; Male ; Mitochondria ; drug effects ; metabolism ; Progesterone ; pharmacology ; Reactive Oxygen Species ; metabolism ; Spermatozoa ; drug effects ; physiology

OBJECTIVETo find a convenient and exact method for evaluating acrosome reaction in human spermatozoa.

METHODSThe semen of the normal male was mixed and then divided into 6 groups. Coomassie brilliant blue (CBB) staining, chlortetracycline (CTC) fluorescence staining and acid phosphatase (ACP) detection were used for morphological observation and data analysis of the acrosome status of the human sperm treated with or without progesterone.

RESULTSThere were obvious morphological differences between the acrosome-reaction and acrosome-intact spermatozoa in CBB staining and CTC fluorescence staining, and significant differences were observed between the experimental and control spermatozoa by the three methods (P < 0.05).

CONCLUSIONAll the three methods can be used to assess acrosome reaction in human spermatozoa, but Coomassie brilliant blue (CBB) staining is much more convenient and stable.

Acid Phosphatase ; Acrosome Reaction ; drug effects ; Cells, Cultured ; Chlortetracycline ; Humans ; Male ; Progesterone ; pharmacology ; Rosaniline Dyes ; Spermatozoa ; cytology ; Staining and Labeling ; methods