Effect of methanol extract of Anacardium occidentale stem bark on agonist-induced contraction in rat uterine muscle

Anacardium occidentale (Ao) is widely used in ethnomedicine and the effect on contractility of the gut and heart muscles of laboratory rodents have been established however, its effect on the contractility of uterine muscle has been rarely researched. The present study investigated the effect of stem bark methanolic extract of Anacardium occidentale (AoME) on Isolated Rat Uterus Strips (IRUS). The Anacardium occidentale methanol extract (AoME) was obtained by solvent extraction. The IRUS were suspended in a bath containing 15 ml of De Jalon's solution through a silk connected to a force transducer. A tension of 1.0 g was applied. The strips were allowed to equilibrate for a period of 60–90 min before the commencement of the experiment. The extract (0.025- 0.3mg/ml) added cumulatively did not modify the normal tonus in most of the IRUS used except in one where it feebly contracted it. However, the contractile responses induced by acetylcholine (10-9 -10-5 M) and oxytocin (10-9 -10-5 M) on the IRUS were almost completely blocked by pre-incubated AoME at a concentration of 0.2 mg/ml. Stem bark methanol extract of Anacardium occidentale shifted the log concentration curve of both agonists to the right. The present results report the antagonistic effects of stem bark methanol extract of Anacardium occidentale on oxytocin and acetylcholine-induced contraction in rat uterus strips. Keywords: Anacardium occidentale, Oxytocin, Acetylcholine, Uterus, Smooth muscle

Ibogaine-Mediated ROS/Antioxidant Elevation in Isolated Rat Uterus Is β-Adrenergic Receptors and KATP Channels Mediated

Ibogaine effects are mediated by cellular receptors, ATP depletion followed by ROS production and antioxidant enzyme activity elevation in a dose and time dependent manner. Since the role of KATP channels and β-adrenoceptors in ROS cellular circuit was established here we explored their role in ibogaine pro-antioxidant effectiveness. Single dose of ibogaine (10 mg/L i.e., 28.8 μmol/L) was applied to isolated rat uterus (spontaneous and Ca2+-stimulated) and contractility and antioxidant enzymes activity were monitored during 4 h. Ibogaine increased amplitude and frequency of spontaneous active uteri immediately after addition that was prevented by propranolol (β1 and β2 adrenoceptors selective antagonists) and glibenclamide (KATP sensitive channels inhibitor; only frequency) pre-treatment. In Ca2+-stimulated uteri, ibogaine decreased both amplitude and frequency after 4 h. Pre-treatment with propranolol abolished ibogaine induced amplitude lowering, while glibenclamide had no effect. In both types of active uterus, ibogaine induced a decrease in SOD1 and an increase in CAT activity after 2 h. In Ca2+-stimulated uterus, there was also a decrease of SOD2 activity after 2 h. After 4 h, SOD1 activity returned to the baseline level, but GSH-Px activity increased. Pre-treatment with both propranolol and glibenclamide abolished observed changes of antioxidant enzymes activity suggesting that ibogaine pro-antioxidative effectiveness is β-adrenergic receptors and KATP channels mediated.

Supplementation with Spirulina platensis Prevents Uterine Diseases Related to Muscle Reactivity and Oxidative Stress in Rats Undergoing Strength Training

Strength training increases systemic oxygen consumption, causing the excessive generation of reactive oxygen species, which in turn, provokes oxidative stress reactions and cellular processes that induce uterine contraction. The aim of this study was to evaluate the possible protective effect of Spirulina platensis (SP), an antioxidant blue algae, on the contractile and relaxation reactivity of rat uterus and the balance of oxidative stress/antioxidant defenses. Female Wistar rats were divided into sedentary (CG), trained (TG), and T + supplemented (TG50, TG100) groups. Reactivity was analyzed by AQCAD, oxidative stress was evaluated by the malondialdehyde (MDA) formation, and the antioxidant capacity was measured by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method. Strength training increased contractile reactivity and decreased the pharmaco-mechanical component of relaxing reactivity in rat uterus. In addition, training decreased oxidation inhibition in the plasma and exercise increased oxidative stress in the uterine tissue; however, supplementation with algae prevented this effect and potentiated the increase in antioxidant capacity. Therefore, this study demonstrated that food supplementation prevents changes in reactivity and oxidative stress induced by strength training in a rat uterus, showing for the first time, that the uterus is a target for this exercise modality and antioxidant supplementation with S. platensis is an alternative means of preventing uterine dysfunction.

A Robust and Highly Efficient Approach for Isolation of Mesenchymal Stem Cells from Wharton’s Jelly for Tissue Repair

BackgroundMesenchymal stem cells (MSCs) derived from umbilical cord Wharton’s Jelly (WJ-MSCs) are emerging as promising therapeutics for a variety of diseases due to their ability of regeneration and immunomodulation and their non-tumorigenic and non-immunogenic properties. Although multiple protocols have been developed for WJ-MSCs isolation, insufficient cell numbers, heterogeneous cell population, and variations in procedures between different laboratories impede further clinical applications.MethodsWe compared six widely used WJ-MSCs isolation methods regarding cell morphology, yield, purity, proliferation rate, and differentiation potential. Based on these analyses, we developed a new isolation approach called “Mince-Soak-Digest (MSD)”, and compared its efficiency with the existing methods. Furthermore, we transplanted WJ-MSCs isolated by different methods to the rat uterus to test their ability for tissue repair.ResultsBased on the comparison and analysis of the six widely used isolation protocols, we identified that the inefficiency of the digestion of the extracellular matrix results in low cell yield. Thus, we have developed a robust and highly efficient method to isolate MSCs from WJ by incorporating a soaking step to facilitate the digestion of the extracellular matrix and release of the cells. Our newly developed method generates significantly higher cell yield (4 to 10-fold higher) than six widely used methods that we tested with high purity and consistency. Importantly, by transplantation of WJ-MSCs isolated by MSD to the rat uterus, we repair the endometrial injury and restore the fertility of the rats.ConclusionOur results provide a robust and highly efficient approach for isolating WJ-MSCs to restore injured tissue. The higher efficiency of MSD assures the abundance of WJ-MSCs for clinical applications. Furthermore, the reliability of MSD contributes to the standardization of WJ-MSCs isolation, which eliminates the discrepancies due to isolation procedures, thus facilitates the evaluation of the efficacy of WJ-MSCs across various human clinical applications.

Determination of potential sources of drug development for menstrual disorders: A qualitative analysis of published literature of in-vitro rat uterus experimental studies

Many herbal plants have been recorded in medicine for their usefulness in menstrual disorders, however, a few have been extensively examined for their pharmacological activities. These plants have been recorded to have usefulness in the management of painful menses, preventing miscarriages, prolonging birth, or inducing birth. Therefore, the effects of herbal plants on the contractility of the uterus will be investigated using the in-vitro experiment of the isolated rat uterus. The study aims to assess the most outstanding plants that are used to treat menstrual disorders, their uterine contractility properties, and adverse effects. The study will therefore acknowledge the importance of medicinal plants in the study of new drugs for regulating uterine contractility and propose suggestions for improving experiments involving medicinal plants on uterine contractility for drug development. The study will provide a direction as to whether the plant extract has drug development potential.

A rat model of male pregnancy

Male pregnancy is a unique phenomenon in syngnathidae which refers to the incubation of embryos or fetuses by males. However, whether male mammalian animals have the potential to conceive and maintain pregnancy remains unclear. Here, we constructed a rat model of male pregnancy by a four-step strategy: a heterosexual parabiotic pair was firstly produced by surgically joining a castrated male rat and a female rat. Uterus transplantation (UTx) was then performed on the male parabiont 8 weeks later. After recovery, blastocyst-stage embryos were transplanted to the grafted uterus of male parabiont and the native uterus of female parabiont. Caesarean section was performed at embryonic day (ED) 21.5. The success rate of modeling was only 3.68%, but 10 pups could still be delivered from male parabionts and developed. Our experiment reveals the possibility of normal embryonic development in male mammalian animals, and it may have a profound impact on reproductive biology.