PGF2α Levels Research Articles

Study of Some Parameters of Recovered Sperm from Vitamin Ctreated Endometriotic Rats Model

The present study was designed to reveal the biochemical and parameters changes of the retrieval sperms uteri of surgically-induced endometriotic rats post 30 min and 1h of coupling. Seventy-five adult female rats were divided into 3 groups as follows: Group 1: 25 shame-operated female rats were orally administrated with 0.1 ml of d.w for 35 days. Group 2: 25 endometriotic female rats were orally administrated with 0.1 ml of d.w for 35 days. Groups 3: 25 endometriotic female rats were orally administrated with 200 mg/Kg of vitamin C for 35 days. At the end of administration, 10 female rats of each groups were mated with fertile males to study sperm parameters. Whereas, others 15 endometriotic female rats were left without mating to use their uteri for leukocytes counts, oxidative markers and PGF2α assay. The retrieval sperms from uteri of endometriotic rats (G2) were showed a significant (p≤0.05) lower in their functions parameters "concentration, viability, progressive motility, and the activity of LDH-C4 enzyme depending on the remaining them in the uteri compared to those values of retrieval sperms from control (G1) and unmated endometriotic rats that treated with vitamin C (G3). Also, the uterine flushing fluids of endometriotic rats (G2) exhibited that had a significant (p≤0.01) increase in the levels of leukocyte counts compared to those values of control (G1) and unmated endometriotic rats with vitamin C (G3). Whereas, there was no significant (P>0.05) difference in the levels of PGF2α in the uterine tissue of all unmated endometriotic groups (G1, G2 and G3). Surgically-induced endometriotic passive influence affected uterine milieu via an appearance of oxidative stress that increased significantly (p≤0.01) as a result of leukocytes count elevation. Ditto, there passive variables in the uterine microenvironment was reflected on the sperm functions parameters leading to infertility. While, the role of vitamin C in ameliorate of all above passive variants was evident.

Physical fitness is associated with prostaglandin F2α isomers during pregnancy

IntroductionPregnancy and physical activity are associated with oxidative stress and immune changes. We hypothesized that pregnant women physically more active in early pregnancy will display a better oxidative stress management and inflammatory response later in pregnancy compared with less active pregnant women. Material and methodsMaternal physical activity using accelerometry monitors for 1 week and cardiorespiratory fitness (VO2 at anaerobic threshold) were assessed at 14–18 weeks in 58 pregnant women. Plasma and erythrocytes membrane samples were obtained from maternal blood samples at 14–18 and 34–37 weeks of pregnancy. Pro-inflammatory prostaglandin (PG) F2α and oxidative stress-derived F2-isoprostanes were measured by high-performance liquid chromatography coupled to tandem mass spectrometry. ResultsHigher physical activity levels at 14–18 weeks measured by mean counts per minute, >30 min/d of moderate to vigorous activity or >6500 steps/d at 14–18 weeks of pregnancy were associated with lower levels of total plasmatic PGF2α later in pregnancy. Concentrations of 5 F2-isomers in erythrocyte membranes in late pregnancy were significantly higher in the third (17.5–19.5 mL kg−1 min−1) and/or fourth (19.6–27.7 mL kg−1 min−1) quartiles of cardio-respiratory fitness compared to the first quartile (13.9–15.9 mL kg−1 min−1). ConclusionsOverall, higher cardio-respiratory fitness in early pregnancy is associated with enhanced erythrocyte membranes oxidation at 34–37 weeks reflecting a higher oxygen transfer capacity. Also, the most active women experienced lower circulating levels of pro-inflammatory PGF2α in plasma at 34–37 weeks, a marker associated with adverse antenatal inflammation-associated conditions. These results support the practice of physical activity by pregnant women.

Lipidomic analysis of lipid mediators derived from cyclooxygenase‐1 and ‐2 pathways reveals their new implications in skeletal muscle

Our published studies showed that treatment with pharmacological inhibitors of cyclooxygenase‐1 (COX‐1) or COX‐2 significantly inhibited myogenesis of C2C12 cells. COX‐1 and ‐2 are rate‐limiting enzymes in the synthesis of lipid mediators, including prostaglandin E2(PGE2), PGF2α, PGD2, PGI2, and thromboxane A2(TXA2), from arachidonic acid (AA). They share some similarities in regulating lipid mediator production, however, previous data suggest that they seem to function differently in the dynamic process of myogenic differentiation.We systemically compared the production of lipid mediators through COX‐1 and ‐2 during myogenesis of mouse primary myoblast using lipidomic analysis to provide new mechanistic insights for their functions in skeletal muscle. Myocytes were transfected with negative control siRNA or specific siRNA against COX‐1 or ‐2 in differentiation medium (DM). 72 h later, medium was collected for lipidomic analysis, and cells were fixed to determine Fusion Index using fluorescent tagged antibody myosin heavy chain.Knockdown of COX‐1 or ‐2 significantly attenuates myogenesis of mouse primary myoblast, demonstrated a significant reduction of by fusion index from 84.8% (negative control) to 74.3% (COX‐1 siRNA) and 73.8% (COX‐2 siRNA). Compared with 0 h of transfection, after 72 h, the levels of PGE2, PGF2α, and 6‐keto‐PGF1α(PGI2stable derivative) in the medium increased. In contrast, during the same period, the levels of PGD2and TXB2(TXA2stable metabolite) decreased. Knocking down COX‐1 significantly reduced the levels of PGE2and PGF2α, but did not alter the levels of 6‐keto‐PGF1α. Knocking down COX‐2 had a similar impact on PGE2levels, but the effect is less pronounced than knocking down COX‐1. Even though the levels of TXB2and PGA2(PGE2 metabolite) at 72 h are lower than at 0 h, after knocking down COX‐1, the concentrations of TXB2and PGA2 significantly decreased compared with control. Moreover, COX‐1 knockdown significantly reduced the release of DHA and OEA from myocytes/myotubes into medium, but COX‐2 knockdown significantly lowered the levels of EPA in culture medium.In addition, reduced COXs activities also affect the levels of AA metabolites through lipoxygenases (LOX) pathways. 5‐ and 12/15 LOXs synthesize 5‐, 12‐, and ‐15‐HETE from AA. We found that reduced COX‐1 or ‐2 expression significantly decreased the levels of 12‐ and 15‐HETE, but no effect on 5‐HETE.In conclusion, COX‐1 could play a predominant role in the production of lipid mediators from AA during mouse primary myoblast myogenesis, and COXs could interact with LOXs to regulate production or signaling of lipid mediators. These results point to a much more complex signaling machinery arising from the regulatory functions of COX‐1 and COX‐2 with implications that can be of long‐term interest for the area of muscle myogenesis and skeletal muscle regeneration. We are now investigating these signaling pathways in animal models and in humans.Support or Funding InformationNIH‐NIA P01 AG039355, the George W. and Hazel M. Jay Endowment (to MB), and the UT System STARS Program (to MB).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Edoxaban improves atrial fibrillation and thromboembolism through regulation of the Wnt-β-induced PI3K/ATK-activated protein C system.

Thromboembolism is a commonly observed condition in geriatrics that is caused by vascular endothelial injury, platelet activation, physiological coagulation processes, reduction of anticoagulant activity, decreased fibrinolytic activity and abnormal flow in the heart chamber, artery or vein. The protein C anticoagulant system serves a crucial role in anticoagulant therapy for the treatment of thromboembolism. Previous findings have suggested that edoxaban is an efficient oral anticoagulant in the acute treatment of venous thromboembolism. In the present study, the efficacy of edoxaban on thromboembolism induced by atrial fibrillation was investigated in a mouse model. Inflammatory factors interleukin (IL)-1, −4, −8 and tumor necrosis factor (TNF)-α were analyzed in the sera of mice with fibrillation induced by thromboembolism. Expression and activity of thymic stromal lymphopoietin (TSLP) and activated protein C resistance were investigated in platelets and vascular endothelial cells (VECs). TSLP-induced platelet viability, Wnt-β phosphorylation and integrin expression were analyzed in platelets. Furthermore, Wnt-β expression and the phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway in VECs were analyzed. Results demonstrated that the expression levels of IL-1, −4, −8 and TNF-α were significantly downregulated in the sera of mice with fibrillation and thromboembolism following treatment with edoxaban (P<0.01). Furthermore, the expression levels of prostacyclin (PGI2), prostaglandin (PG)E2, PGD2 and PGF2α were significantly increased in the sera of experimental mice that received edoxaban therapy (P<0.01). Results also indicated that edoxaban significantly stimulated the protein expression of TSLP and activated Wnt-β phosphorylation and integrin expression in platelets (P<0.01). In addition, edoxaban therapy significantly upregulated the expression levels of PI3K and AKT, and subsequently increased the activity of protein C and S in VECs (P<0.01). Notably, edoxaban treatment improved atrial fibrillation and thromboembolism, as determined by pathological analysis. In conclusion, these results suggested that edoxaban elicited beneficial effects for mice with atrial fibrillation induced by thromboembolism through the regulation of the Wnt-β-induced PI3K/ATK-activated protein C system.

Herbal-cake-partitioned moxibustion of "Shenque" (CV8) has a relative specific effect in relieving abdominal pain and in regulating neuroendocrine-immune network in primary dysmenorrhea rats

To observe the effect of herbal-cake-partitioned moxibustion (HCPM) of "Shenque" (CV8) and "Daheng" (SP15) on abdominal pain, plasma β-endorphin (β-EP), uterine prostaglandin E2 (PGE2) and prostaglandin F2α (PGF2α) levels, as well as splenetic natural killer cell (NK cell) activity in primary dysmenorrhea (PD) rats, so as to explore the specificity of acupoint function and the underlying mechanisms of moxibustion in relieving dysmenorrhea. A total of 40 female rats were randomized into blank control, model, CV8-direct moxibustion, CV8-HCPM and SP15-HCPM groups (n＝8 rats in each). The PD model was established by subcutaneous injection of estradiol benzoate injection (0.2-0.5 mg/rat) for 10 consecutive days and intraperitoneal injection of oxytocin (2 U) 24 h after the last subcutaneous injection. Moxibustion or herbal-cake (composed of Radix Angelicae Sinensis, Rhizoma Chuanxiong, Radix Paeoniae Rubra, Cortex Cinnamomi, etc.)-partitioned moxibustion was applied to CV8, SP15 or umbilicus respectively for 7 moxa-cones every time, once daily for 10 successive days. The rats of the control and model groups were also restrained as those in the moxibustion groups. The writhing times within 30 minutes was recorded and the contents of plasma β-EP, uterine PGE2 and PGF2α were detected by ELISA, and NK cell activity was detected using MTT. Compared with the control group, the writhing times and the content of PGF2α in the uterus tissue were significantly increased in the model group (P<0.01), while the contents of plasma β-EP, uterine PGE2 and splenetic NK cell activity were significantly decreased (P<0.01). In comparison with the model group, the writhing times and uterine PGF2α content were obviously down-regulated in the SP15-HCPM, CV8-direct moxibustion and CV8-HCPM groups (P<0.05, P<0.01), and the contents of plasma β-EP and uterine PGE2, and splenetic NK cell activity were significantly increased (P<0.05, P<0.01). The therapeutic effects of CV8-HCPM group were significantly superior to those of SP15-HCPM and CV8-direct moxibustion groups in lowering writhing times and PGF2α level, and in up-regulating β-EP, PGE2 (P<0.05, P<0.01). The NK cell activity of CV8-HCPM group was significantly increased compared with that in the SP15-HCPM group(P<0.05). No significant differences were found between the SP15-HCPM and CV8-direct moxibustion groups in the levels of writhing times, plasma β-EP and uterine PGE2, PGF2α contents and splenetic NK cell activity (P>0.05). Moxibustion of both CV8 and SP15 can relieve abdominal pain in PD rats, which may be closely associated with its effect in suppressing PD-induced decrease of plasma β-EP and uterine PGE2 levels and splenetic NK cell activity and increase of uterine PGF2α. The therapeutic effect of CV8-HCPM is obviously better than that of SP15-HCPM and CV8-direct moxibustion.

Prostaglandins E2 and F2α levels in human menstrual fluid by online Solid Phase Extraction coupled to Liquid Chromatography tandem Mass Spectrometry (SPE-LC-MS/MS)

This paper reports an online SPE-LC-MS/MS method for the simultaneous quantification of prostaglandins (PGE2 and PGF2α) in menstrual fluid samples. To meet this goal human peripheral serum was used as surrogate matrix. The analytes were trapped on an OASIS HLB cartridge for 3 min, for sample cleanup and enrichment, and then transferred during only 42 s to an HSS T3 C18 analytical column, for separation and analysis. Prostaglandins (PGs) were detected by selected reaction monitoring in negative ion mode, PGE2 (m/z 351 → 315) and PGF2α (m/z 353 → 193) using isotope-labeled internal standard (PGE2-d4, m/z 355 → 319). The concentration linear range was of 10.34–1.034 ng mL−1 and the lower limit of quantification (LLOQ) was 10.34 ng mL−1 for both PGs. Validation parameters were successfully assessed according to the European Medicines Agency guideline (EMA), also comprising the FDA normative. The method showed no matrix effect and process efficiency around 100%, in addition to only 15 min of analysis time with lower solvent consumption. The method application was carried out using two menstrual fluid sample groups: control (n = 15) and treatment group (n = 7; samples from women that used Tahiti lemon juice). The PGF2α levels were found to be higher in treated group than in control group (p ≤ 0.05), denoting an effect of the intake of Tahiti lemon juice on the menstrual inflammatory process. The on-line method herein reported could be useful for the analysis of PGs from large research studies.

90 Effects of treatment with the nonsteroidal anti-inflammatory drug Anafen prior to embryo recovery and the number of embryo flush procedures performed on prostaglandin levels in uterine fluid and pregnancy rate following embryo transfer

Some studies have shown that performing a second flush during embryo collection can increase the number of embryos recovered. However, this technique results in greater uterine manipulation that may increase PGF2α in the uterine fluid of donor cows and affect pregnancy rate in recipient females. Two experiments were conducted (1) to determine whether PGF2α concentration in uterine fluid was affected by treatment with a non-steroidal anti-inflammatory drug (NSAID) before embryo recovery and (2) to determine whether the percent of recipients becoming pregnant following embryo transfer (P/ET) was affected by flush number. In both experiments, lactating Holstein cows were used as donors (n=18 and 34, respectively) and subjected to a standard superovulation protocol (Baracaldo et al. 2000 Theriogenology 53, 1239-1250, DOI: 10.1016/S0093-691X(00)00268-5). Embryo recovery was performed using 2 catheters that were inserted simultaneously into the uterus, 1 into each horn. During the first flush, a volume of 300mL of flush medium was used in each horn. At the end of the first flush approximately 75mL of fluid was left and, after 1h, a second flush was performed to recover the remaining fluid. Samples were taken from each horn at the first and second flush to determine PGF2α concentration using a bovine PGFM ELISA kit. Data for PGF2α concentrations in uterine fluid were presented as the average of both uterine horns. In Exp. 1, donors were randomly assigned to be treated with or without 1.2g of the NSAID Anafen (ketoprofen) IM 20min before embryo recovery. In Exp. 2, recovered grade-1 embryos from the first and second flush procedures were frozen in ethylene glycol and subsequently transferred to synchronized Holstein heifers (n=106 and 49 for the first and second flushes, respectively). Pregnancy was diagnosed by rectal palpation at Day 60. All data were analysed using t-test. In Exp. 1, Anafen treatment 20min before embryo recovery reduced PGF2α in uterine fluid during the second flush procedure (105.9±11.4v. 43.6±5.7 pg/mL; P&amp;lt;0.001), but not during the first (54.7±7.3v. 44.2±5.6 pg/mL; P=0.34). In Exp. 2, PGF2α in uterine fluid was greater for the second flush as compared to the first flush (95.9±7.4v. 56.3±5.5 pg/mL; P&amp;lt;0.001). A total of 345 viable embryos were recovered from the 34 donors in Exp. 2 (n=276 and 69 for the first and second flushes, respectively). The P/ET at Day 60 was greater (P&amp;lt;0.05) for recipients that received embryos from the first flush as compared to recipients that received embryos from the second flush (64.2v. 49.0%). Taken together, the results of the present study indicate that the second flush procedure results in greater PGF2α levels in uterine fluid and that embryos recovered during the second flush are less likely to establish pregnancy following transfer. Moreover, pretreatment of donors with the NSAID Anafen reduced PGF2α levels in uterine fluid during the second flush. Further research is needed to determine whether treatment of donors with NSAID can improve the P/ET achieved following the transfer of embryos recovered following a second flush procedure.

Endometrial prostaglandin F2α in vitro production and its modulation regarding dominant follicle position in cattle

Prostaglandin F2α (PGF2α) determines luteolysis in cattle, and the ability to manipulate its endogenous synthesis is indispensible for large-scale animal breeding. Estradiol (E2) and progesterone (P4) modulate several molecular pathways in endometrial cells, including the synthesis of PGF2α; however, its specific mechanisms are still not totally known. This study investigated the production in vitro and possible modulation of endometrial PGF2α due to a local effect of endogenous E2 in the ipsilateral uterine horn (UH) containing the dominant follicle (DF) or from P4 in ipsilateral horn containing the corpus luteum (CL). The PGF2α stimulators oxytocin (OT) and phorbol 12,13-dibutyrate (PDBu) were incubated with endometrial explants, and PGF2α content was measured. For that, cycling cows were synchronized, the development of DF and CL was examined by ultrasonography and on the seventh day of the estrous cycle, endometrial explants were collected and cultured in medium supplemented with 10-6 M PDBu or 10-6 M OT or non-supplemented. Media samples were collected immediately after treatment and 60 min later. Radioimmunoassay showed that the PGF2α content of the UH ipsilateral to the DF was 49% less than that of the contralateral UH (8.22 ± 0.95 vs. 12.24 ± 0.95 pg/mL/mg tissue, respectively; P &lt; 0.01). However, the PGF2α levels did not differ between the UHs as a function of the CL position (9.46 ± 0.95 vs. 11 ± 0.95 pg/mL/mg; P &gt; 0.05). The cellular stimulators promoted an increase in PGF2α synthesis (P &lt; 0.02), and the effects differed among the animals (P &lt; 0.04). The PGF2a production was higher in the explants treated with PDBu rather than OT (13.68 ± 1.16 vs. 10.01 ± 1.16 pg/mL/mg tissue, respectively; P &lt; 0.05). In conclusion, PGF2α synthesis is modulated by the presence of the DF (local E2) but not the CL (local P4), and both PDBu and OT stimulated PGF2a synthesis.

F2-isoprostanes and fatty acids profile in early pregnancy complicated by pre-existing diabetes

BackgroundDiabetes and pregnancy are both associated with oxidative stress, characterized by an increase of F2-isoprostanes from the non-enzymatic oxidation of arachidonic acid, a n − 6 polyunsaturated fatty acid (PUFA). We hypothesized that pregnant women with pre-existing diabetes will be characterized by elevated levels of specific F2-isoPs isomers and altered PUFA composition in plasma early pregnancy when compared to normoglycemic controls. MethodsPlasma samples from 23 women with uncomplicated pregnancies and 11 women with pre-existing diabetes in pregnancy were collected between 12 and 18 weeks of pregnancy (MIROS cohort). Six F2-isoprostanes isomers were measured by high-performance liquid chromatography coupled to tandem mass spectrometry. Fatty acids concentrations in plasmatic phospholipids were measured by gas chromatography coupled to a flame ionization detector. ResultsF2-isoprostanes, specifically the 8-iso-15(R)-PGF2α levels, were 67% higher in diabetic than normoglycemic pregnancies (p = 0.026). The total n − 6 PUFA and arachidonic acid level did not differ between study groups. In contrast, total n − 3 level was 32% lower in diabetic pregnancies than in controls (p = 0.002); EPA(20:5) and DHA(22:6) being specifically reduced (p = 0.035 and p = 0.003 respectively). Delta-6-desaturase (D6D) activity index, calculated using fatty acid ratios, was 9% lower in pre-existing diabetes than in controls (p = 0.042). ConclusionsPre-existing diabetes in early pregnancy displays a distinctive F2-isoprostanes profile when compared to other pathologies of pregnancy, such as preeclampsia, as previously assessed in the same cohort.

High levels of circulating prostaglandin F2α associated with ovulation stimulate female sexual receptivity and spawning behavior in the goldfish (Carassius auratus)

This study tested the hypothesis that blood-borne prostaglandin F2α (PGF2α) produced at the time of ovulation by female goldfish, a typical scramble-spawning, egg-laying cyprinid fish, functions as a hormone which stimulates female sexual receptivity, behavior, and pheromone release, thereby synchronizing female mating behavior with egg availability. We conducted 5 experiments. First, we tested whether PGF2α is found in the blood of female fish and if it increases at the time of ovulation. Using gas chromatography–mass spectrometry, we found that circulating PGF2α was approximately 1 ng/ml prior to ovulation, increased over 50-fold within 3 h of ovulation and returned to preovulatory values after spawning and egg release. Ovulated fish also released over 2 ng/h of PGF2α and 800 ng/h of 15-keto-PGF2α, a metabolite of PGF2α – both compounds with known pheromonal function. Second, we tested how closely levels of circulating PGF2α tracked the timing of ovulation by sampling fish at the time of ovulation and discovered that PGF2α increased within 15 min of ovulation, peaked after 9 h, and fell to basal levels as fish spawned and released their eggs. Third, we tested whether an interaction between eggs and the reproductive tract serves as a source of circulating PGF2α and its relationship with female sexual receptivity by injecting ovulated eggs (or an egg-substitute) into the reproductive tract of females stripped of ovulated eggs. We found both of these treatments elicited measurable increases in plasma PGF2α as well as female sexual behavior. A fourth experiment showed that indothemacin, a PG synthase inhibitor, blocked both PGF2α increase and female sexual behavior in egg-substitute-injected fish. Finally, we tested the relationship between the expression of female behavior and PGF2α in PGF2α-injected fish and found that circulating PGF2α levels closely paralleled behavior, rising within 15 min and peaking at 45 min. Together, these experiments establish that PGF2α functions as a behavioral blood-borne hormone in the goldfish, suggesting it likely has similar activity in other related, externally-fertilizing fishes.

Attracting Common Carp to a bait site with food reveals strong positive relationships between fish density, feeding activity, environmental DNA, and sex pheromone release that could be used in invasive fish management.

Measurement of environmental DNA (eDNA) is becoming a common technique to survey for rare and invasive fish due to its sensitivity and specificity. However, its utility is limited by an incomplete understanding of factors governing its sources and fates. Failure to detect eDNA is especially difficult to interpret so surveillance techniques often collect large numbers of samples across broad regions. If, however, fish could be reliably attracted to a single location where their eDNA could be easily measured that would be useful. We conducted a proof‐of‐concept study of this idea using invasive Common Carp. We monitored the distribution of radio‐tagged Carp and their eDNA across a 67 ha lake focusing at the bait site while a pheromone (Prostaglandin F2α; PGF 2α) was also measured to determine their reproductive condition. Prior to baiting, Carp were patchily distributed and while eDNA was occasionally detectable, it was patchy and only loosely associated with moderately dense groups of fish. Further, neither Carp, nor their eDNA were consistently measurable at the bait site and surrounding region, and the pheromone was not measurable at all. However, once baiting commenced, Carp started visiting the bait site and feeding, especially at night, where eDNA levels increased 500‐fold as fish densities doubled and PGF 2α became detectable. Fish presence, eDNA and pheromone concentrations peaked at night after 6 days, strongly suggesting feeding activity was the main driver. While the presence of eDNA precisely coincided with this aggregation, levels had dropped dramatically within 5 m. PGF 2α levels dropped less rapidly and demonstrated the presence of live mature fish. We suggest that food could be used to train fish to come to locations where they otherwise are too scarce to be reliably measured, increasing their eDNA release, making them measurable, and their reproductive condition also discernable by measuring pheromones.

Prostaglandin F2α Receptor Modulation Affects Eye Development in Guinea Pigs.

Retinal arachidonic acid (ARA) levels in form-deprived eyes decline in guinea pigs. As prostaglandin F2α (PGF2α) is an ARA metabolite and endogenous agonist of prostaglandin F receptor (FP), we have been suggested that down-regulation of PGF2α-FP receptor signalling pathway contributes to myopia onset. To test this hypothesis, this study determines whether: (i) retinal PGF2α levels decline during the development of form deprivation myopia (FDM) in guinea pigs; (ii) FP receptor agonism and antagonism alter emmetropization and myopia development. Pigmented guinea pigs were randomly assigned to normal vision and form-deprived groups. Ultraperformance liquid chromatography coupled with a mass spectrometer (UPLC-MS) measured retinal PGF2α levels 2weeks after form deprivation (FD). The selective FP agonist, latanoprost acid (LAT) and its corresponding antagonist, AL8810, were peribulbarly injected into each group. An eccentric infrared photorefractor (EIR) monitored refraction. A-scan ultrasonography measured axial elongation (AL) and vitreous chamber depth (VCD). Tonometry measured the intraocular pressure (IOP). Retinal PGF2α levels declined in form-deprived eyes compared to those in normal eyes. Neither LAT nor AL8810 affected IOP with or without FD. On the other hand, after 4weeks of daily 0.5μg AL8810 treatment, a myopia of -1.99±0.34dioptre (D) developed, but LAT had no effect on emmetropization in a normal visual environment. Nevertheless, daily 30μg LAT treatment for 4weeks inhibited FDM development by 41% (vehicle control: -8.39±0.45D; LAT: -4.95±0.39D; two-way anova with repeated measures, p<0.05). Down-regulation of PGF2α-FP receptor signalling pathway may contribute to myopia onset as retinal PGF2α declined in myopic eyes and antagonism of FP receptor by AL8810 induced a myopic shift in normal vision environment. Meanwhile, up-regulation of this pathway by LAT inhibited FDM development. However, the mechanism underlying LAT-induced FDM inhibition needs further clarification. This uncertainty exists because its inhibition of FDM suggests that LAT strengthens the scleral framework which reduces axial elongation. On the other hand, its IOP-lowering effect is attributed to thinning and weakening the scleral framework in glaucoma treatment.

Extracts of compound Muniziqi granule suppressed uterus contraction and ameliorated oxytocin-induced primary dysmenorrhea

Ethnopharmacological relevanceCompound Muniziqi granule (CMG) is usually used as a traditional Uighur medicine to treat acne, chloasma, skin inflammation, primary dysmenorrhea (PDM), and menopausal syndrome. However, there are no sufficient data to support the clinic uses of CMG in PDM. Aim of the studyThis work aims to examine the effect of CMG as a treatment for PDM and reveal its possible therapeutic mechanism. Materials and methodsIn vivo and in vitro mouse PDM models were utilized in this study. The mouse uterine contraction was induced by oxytocin after progynova or estradiol benzoate pretreatment. CMG, alkaloid extracts from seeds of Peganum harmala (AEP), and 10% and 95% ethanol extracts from seeds of Nigella glandulifera (EEN10 and EEN95) were given to mice in three doses by gavage. The writhing times within 30 min after oxytocin treatment were recorded to evaluate the analgesic effect, and the glutathione peroxidase (GSH-Px), malondialdehyde (MDA), 6-keto-prostaglandin F1α (6-k-PGF1α), prostaglandin F2α (PGF2α), thromboxane B2 (TXB2), and nitric oxide (NO) levels in uterine tissues and PGF2α and MDA in serum were determined. The effects (contractile curve) of CMG, AEP, EEN10, and EEN95 on uterus contraction induced by oxytocin in isolated mouse uterus were recorded. ResultsIn contrast to the control group, CMG, AEP, N10, and N95 could display analgesic activities dose dependently by reducing the writhing response of the PDM model mice. CMG, AEP, EEN10, and EEN95 could also remarkably decrease the level of PGF2α, 6-k-PGF1α, TXB2, NO and MDA in uterine tissues and PGF2α and MDA in serum, whereas the activity of GSH-Px in uterine tissues was increased. Furthermore, CMG, AEP, EEN10, and EEN95 could significantly inhibit the frequency and amplitude of isolated uterus induced by oxytocin in a concentration-dependent manner. ConclusionsCMG exhibited a significant protective effect on experimental PDM. The mechanisms are probably associated with abating lipid peroxidation and over-inflammatory reaction, and alleviating the contraction of isolated mouse uterus. The seeds of P. harmala and N. glandulifera in the CMG may play an important role in exerting protective effects on PDM. This study provides pre-clinic proof to the use of CMG in clinical practice of PDM.

PGF2α and gonadal steroid plasma levels of successful and unsuccessful spawning Piaractus mesopotamicus (Teleostei, Characiformes) females

Gonadal steroid and prostaglandin F2α (PGF2α) plasma levels were evaluated in successfully (SP) and unsuccessfully ovulated (UN) female Piaractus mesopotamicus. Forty-one females were injected with crude carp pituitary extract (0.6 and 5.4 mg kg−1 with a 24-h interval between the doses) and sampled to determine the plasma concentration of 17β-estradiol (E2), 17α-hydroxyprogesterone (17α-OHP), 17α,20β-dihydroxy-4-pregnen-3-one (DHP), PGF2α, and testosterone (T) after each injection (first—A1 and second—A2), and at the time of ovulation for SP and UN. Two clusters were obtained using multivariate analysis: 1—composed of all A1, all A2, and some UN; and 2—composed of all SP and some UN. Median values of E2 plasma levels were similar between clusters; however, plasma levels of T, 17α-OHP, DHP, and PGF2α of cluster 2 (predominantly formed by SP) were higher than those of cluster 1. Since cluster 2 contained all SP and females of this cluster presented higher levels of PGF2α, T, 17α-OHP, and DHP, here we evidently shown in an unprecedented manner that concomitant increased levels of these substances were associated with successful ovulation in this species, but such an increase was not determinant for successful ovulation due to the presence of some UN females in the same cluster 2. These findings highlight the unexplored potential of PGF2α to be used as an accessory tool for inducing successful ovulation for fish farming purposes.

Evaluation of mitochondrial redox status and energy metabolism of X-irradiated HeLa cells by LC/UV, LC/MS/MS and ESR

To evaluate the metabolic responses in tumour cells exposed to ionizing radiation, oxygen consumption rate (OCR), cellular lipid peroxidation, cellular energy status (intracellular nucleotide pool and ATP production), and mitochondrial reactive oxygen species (ROS), semiquinone (SQ), and iron–sulphur (Fe−S) cluster levels were evaluated in human cervical carcinoma HeLa cells at 12 and 24 h after X-irradiation. LC/MS/MS analysis showed that levels of 8-iso PGF2α and 5-iPF2α-VI, lipid peroxidation products of membrane arachidonic acids, were not altered significantly in X-irradiated cells, although mitochondrial ROS levels and OCR significantly increased in the cells at 24 h after irradiation. LC/UV analysis revealed that intracellular AMP, ADP, and ATP levels increased significantly after X-irradiation, but adenylate energy charge (adenylate energy charge (AEC) = [ATP + 0.5 × ADP]/[ATP + ADP + AMP]) remained unchanged after X-irradiation. In low-temperature electron spin resonance (ESR) spectra of HeLa cells, the presence of mitochondrial SQ at g = 2.004 and Fe–S cluster at g = 1.941 was observed and X-irradiation enhanced the signal intensity of SQ but not of the Fe–S cluster. Furthermore, this radiation-induced increase in SQ signal intensity disappeared on treatment with rotenone, which inhibits electron transfer from Fe–S cluster to SQ in complex I. From these results, it was suggested that an increase in OCR and imbalance in SQ and Fe–S cluster levels, which play a critical role in the mitochondrial electron transport chain (ETC), occur after X-irradiation, resulting in an increase in ATP production and ROS leakage from the activated mitochondrial ETC.

Impact of dietary ω3 polyunsaturated fatty acid supplementation on brown and brite adipocyte function

The recent characterization of functional brown adipose tissue in adult humans has opened new perspectives for regulation of energy expenditure with respect to obesity and diabetes. Furthermore, dietary recommendations have taken into account the insufficient dietary intake of ω3 PUFAs and the concomitant excessive intake of ω6 PUFA associated with the occurrence of overweight/obesity. We aimed to study whether ω3 PUFAs could play a role in the recruitment and function of energy-dissipating brown/brite adipocytes. We show that ω3 PUFA supplementation has a beneficial effect on the thermogenic function of adipocytes. In vivo, a low dietary ω6:ω3 ratio improved the thermogenic response of brown and white adipose tissues to β3-adrenergic stimulation. This effect was recapitulated in vitro by PUFA treatment of hMADS adipocytes. We pinpointed the ω6-derived eicosanoid prostaglandin (PG)F2α as the molecular origin because the effects were mimicked with a specific PGF2α receptor agonist. PGF2α level in hMADS adipocytes was reduced in response to ω3 PUFA supplementation. The recruitment of thermogenic adipocytes is influenced by the local quantity of individual oxylipins, which is controlled by the ω6:ω3 ratio of available lipids. In human nutrition, energy homeostasis may thus benefit from the implementation of a more balanced dietary ω6:ω3 ratio.

In Vitro and In Vivo Antioxidant Activities of the Flowers and Leaves from Paeonia rockii and Identification of Their Antioxidant Constituents by UHPLC-ESI-HRMSn via Pre-Column DPPH Reaction.

The genus Paeonia, also known as the “King of Flowers” in China, is an important source of traditional Chinese medicine (TCM). Plants of this genus have been used to treat a range of cardiovascular and gynecological diseases. However, the potential pharmacological activity of one particular species, Paeonia rockii, has not been fully investigated. In the first part of the present study, 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis-(3-ethylbenzothiazoline-6-sulfonic) acid (ABTS), reducing power assays, and metal ion chelating assays were used to investigate the in vitro antioxidant activities of Paeonia rockii. In the second portion of the study, a mouse model of d-galactose-induced aging was used to validate the antioxidant effects of the flowers from Paeonia rockii in vivo. Lastly, potential antioxidant constituents were screened and identified by ultra-high pressure liquid chromatography and electrospray ionization coupled with high-resolution mass spectrometry (UHPLC-ESI-HRMSn) combined with the DPPH assay. Results indicated that the flowers and leaves exhibited stronger antioxidant activity than ascorbic acid in vitro. The therapeutic effect of Paeonia rockii was determined in relation to the levels of biochemical indicators, such as 8-iso-prostaglandin F2α (8-iso PGF2α) in the serum, superoxide dismutase (SOD), protein carbonyl, malondialdehyde (MDA), and glutathione (GSH) in the liver and brain, after daily intra-gastric administration of different concentrations of extracts (100, 200 and 400 mg/kg) for three weeks. The levels of 8-iso PGF2α (p < 0.01) and protein carbonyl groups (p < 0.01) were significantly reduced, whereas those of SOD (p < 0.05) had significantly increased, indicating that components of the flowers of Paeonia rockii had favorable antioxidant activities in vivo. Furthermore, UHPLC-ESI-HRMSn, combined with pre-column DPPH reaction, detected 25 potential antioxidant compounds. Of these, 18 compounds were tentatively identified, including 11 flavonoids, four phenolic acids, two tannins, and one monoterpene glycoside. This study concluded that the leaves and flowers from Paeonia rockii possess excellent antioxidant properties, highlighting their candidacy as “new” antioxidants, which can be utilized therapeutically to protect the body from diseases caused by oxidative stress.

Effects of electroacupuncture on luteal regression and steroidogenesis in ovarian hyperstimulation syndrome model rat

AimsElectroacupuncture (EA) is an effective and safe therapeutic method widely used for treating clinical diseases. Previously, we found that EA could decrease serum hormones and reduce ovarian size in ovarian hyperstimulation syndrome (OHSS) rat model. Nevertheless, the mechanisms that contribute to these improvements remain unclear. Materials and methodsHE staining was used to count the number of corpora lutea (CL) and follicles. Immunohistochemical and ELISA were applied to examine luteal functional and structural regression. Immunoprecipitation was used for analyzing the interaction between NPY (neuropeptide Y) and COX-2; western blotting and qRT-PCR were used to evaluate the expressions of steroidogenic enzymes and PKA/CREB pathway. Key findingsEA treatment significantly reduced the ovarian weight and the number of CL, also decreased ovarian and serum levels of PGE2 and COX-2 expression; increased ovarian PGF2α levels and PGF2α/PGE2 ratio; decreased PCNA expression and distribution; and increased cyclin regulatory inhibitor p27 expression to have further effect on the luteal formation, and promote luteal functional and structural regression. Moreover, expression of COX-2 in ovaries was possessed interactivity increased expression of NPY. Furthermore, EA treatment lowered the serum hormone levels, inhibited PKA/CREB pathway and decreased the expressions of steroidogenic enzymes. Hence, interaction with COX-2, NPY may affect the levels of PGF2α and PGE2 as well as impact the proliferation of granulosa cells in ovaries, thus further reducing the luteal formation, and promoting luteal structural and functional regression, as well as the ovarian steroidogenesis following EA treatment. SignificanceEA treatment could be an option for preventing OHSS in ART.

Effects of the Acupoint Catgut Embedding on Nerve-Endocrine-Immune Network in Dysmenorrhea Rats

To explore the underlying mechanism of acupoint catgut embedding in improving primary dysme-norrhea (PD) in rats based on functional activities of the neuro-endocrine-immune (NEI) network. Forty female rats were equally randomized into blank control, PD model, medication, and acupoint catgut embedding groups. The PD model was established by subcutaneous injection of estradiol benzoate (0.5 mg/rat on the 1st and 10th d, and 0.2 mg/rat from 2nd to 9th d) and oxytocin (2 U/rat, i．p.). Rats of the medication group were treated by intragastric perfusion of fenbid (0.8 mL/rat, 125 mg/100 mL), once daily for 10 days. The catgut embedding was applied to bilateral "Ciliao" (BL 32), "Sanyinjiao" (SP 6) and "Guanyuan" (CV 4) before modeling. The body writhing times in 30 minutes were recorded, plasma β-endorphin(β-EP) content, and prostaglandin E 2 (PGE2) and prostaglandin F 2 α (PGF2α) contents in the uterus tissue were assayed using ELISA, and the activity of natural killer cell (NK cell) in the spleen tissue was detected using 3-(4，5-dimethyl-2-thiazolyl)-2，5-diphenyl-2-H-tetrazolium bromide (MTT) method after isolation and co-culture with K 562 cells. The body writhing times were no-tably more in the model group than in the control group (P<0.01), and obviously fewer in both medication and catgut embedding groups than in the model group (P<0.01). After modeling, the plasma β-EP and uterus PGE2 contents and splenic NK cell activity were significantly decreased (P<0.01), while the uterus PGF2α content was evidently increased in the model group relevant to the control group (P<0.01). Following the treatment, plasma β-EP and uterus PGE2 contents and splenic NK cell activity were considerably up-regulated (P<0.01), and uterus PGF2α content was markedly down-regulated in both medication and acupoint catgut embedding groups (P<0.01), suggesting an involvement of the NEI network in catgut embedding-induced improvement of PD. The therapeutic effect of catgut embedment was markedly superior to that of medication in up-regulating splenic NK cell activity (P<0.01). No significant differences were found between the medication and catgut embedding groups in the body writhing times within 30 min, and in the levels of plasma β-EP and uterus PGE2 and PGF2α (P>0.05). The acupoint catgut embedding has a significant efficacy in relieving PD in rats, which may be related to its effect in up-regulating plasma β-EP, uterus PGE2 contents and splenic NK cell activity and in down-regulating uterus PGF2α level.

Heat stress affects prostaglandin synthesis in bovine endometrial cells

Heat stress (HS) negatively affects reproduction in cattle; however, its effect on endocrine function in bovine endometrial cells remains unclear. In this study, we examined the effects of HS on the production of prostaglandin (PG) E2 and PGF2α in the cultured bovine endometrial epithelial and stromal cells separately. To evaluate the effect of HS on endocrine function, the cells were cultured at 38.5°C (control) or 40.5°C (HS). After treatment, PGE2 and PGF2α levels were measured via enzyme immunoassay (EIA) and mRNA expressions of enzymes involved in PG synthesis were examined via quantitative reverse transcription polymerase chain reaction (RT-PCR). HS did not influence the production of PGE2 or PGF2α in the epithelial cells; however, HS significantly enhanced the production of both PGE2 and PGF2α in the stromal cells (P < 0.05). In addition, HS significantly increased phospholipase A2 (PLA2), cyclooxygenase 2 (COX2), prostaglandin F synthase (PGFS), prostaglandin E synthase (PGES), and carbonyl reductase 1 (CBR1) mRNA expression in the stromal cells (P < 0.05). The overall results suggest that HS induces mRNA expression of enzymes involved in PG synthesis, resulting in the upregulation of PGE2 and PGF2α production in the stromal cells, but not in the epithelial cells. The HS-induced increase of PGE2 and PGF2α secretion in bovine endometrial stromal cells may disrupt the normal estrous cycle and cause infertility in cows during summer.