Female SD Research Articles

Differential Effects of Clozapine and Haloperidol on the 40 Hz Auditory Steady State Response-mediated Phase Resetting in the Prefrontal Cortex of the Female Sprague Dawley Rat.

Neural synchrony at gamma frequency (~40 Hz) is important for information processing and is disrupted in schizophrenia. From a drug development perspective, molecules that can improve local gamma synchrony are promising candidates for therapeutic development. Given their differentiated clinical profile, clozapine, and haloperidol may have distinct effects on local gamma synchrony engendered by 40 Hz click trains, the so-called auditory steady-state response (ASSR). Clozapine and haloperidol at doses known to mimic clinically relevant D2 receptor occupancy were evaluated using the ASSR in separate cohorts of female SD rats. Clozapine (2.5-10 mg/kg, sc) robustly increased intertrial phase coherence (ITC), across all doses. Evoked response increased but less consistently. Background gamma activity, unrelated to the stimulus, showed a reduction at all doses. Closer scrutiny of the data indicated that clozapine accelerated gamma phase resetting. Thus, clozapine augmented auditory information processing in the gamma frequency range by reducing the background gamma, accelerating the gamma phase resetting and improving phase precision and signal power. Modest improvements in ITC were seen with Haloperidol (0.08 and 0.24 mg/kg, sc) without accelerating phase resetting. Evoked power was unaffected while background gamma was reduced at high doses only, which also caused catalepsy. Using click-train evoked gamma synchrony as an index of local neural network function, we provide a plausible neurophysiological basis for the superior and differentiated profile of clozapine. These observations may provide a neurophysiological template for identifying new drug candidates with a therapeutic potential for treatment-resistant schizophrenia.

Water tank scale: a reliable method for assessing motor function after spinal cord injury in rats

To analyze the reliability of the Water Tank Scale for assessing recovery of motor function after spinal cord injury (SCI) in rats. Thirty-six adult female SD rats were randomly divided into SCI and sham-operated groups (n= 18). The recovery of the hind limb motor function was assessed using Water Tank scoring, BBB scoring, and motor-evoked potentials (MEP) at 1, 3, 5, 7, 14 and 21 days after SCI. MEP was used as the gold standard for analyzing and comparing differences between the two scoring methods. The Water Tank scores of the rats were significantly higher than the BBB scores on day 3 (0.22±0.43 vs 0, P < 0.05) and also on days 5, 7 and 14 after SCI (0.67±0.49 vs 0.11±0.32, 4.33±1.19 vs 2.83±1.04, 8.61± 1.20 vs 7.06±1.0, P < 0.01). On day 21 after SCI, the scores of the Water Tank Scale of the rats did not significantly differ from the BBB scores (14.78±1.06 vs 14.50±1.47, P>0.05). Neurophysiological monitoring showed that both the Water Tank score and BBB score were significantly correlated with MEP latency, but the Water Tank score had a greater correlation coefficient with MEP latency (r=-0.90). Compared with the BBB scale, Water Tank scoring allows more objective and accurate assessment of functional recovery of the spinal cord in early stages following SCI in rats, and can thus be used as a reliable method for assessing functional recovery of the hind limbs in rat models of acute SCI.

Prophylactic Effects of Hemp Seed Oil on Perimenopausal Depression: A Role of HPA Axis.

Hemp seed, the dried fruit of Cannabis sativa L. (Moraceae), has been extensively documented as a folk source of food due to its nutritional and functional value. This study evaluated the antidepressant effect of hemp seed oil (HSO) during its estrogen-like effect in Perimenopausal depression (PMD) rats induced by ovariectomy combined with chronic unpredictable mild stress (OVX-CUMS). Female SD rats (SPF, 10 weeks, sham operated group, ovariectomy (OVX) model group, ovariectomy - chronic unpredictable mild stress (OVX-CUMS) group, HSO + OVX-CUMS group, fluoxetine (FLU) + OVX-CUMS group, n=8) were subjected to treatment with HSO (4.32 g/kg) or fluoxetine (10 mg/kg) for 28 days (20 mL/kg by ig). Sucrose preference test (SPT), forced swimming test (FST), open field test (OFT), estrogen receptor α (ERα) and estrogen receptor β (ERβ) expression, estradiol (E2), follicle stimulating hormone (FSH), luteinizing hormone (LH), cortisol (CORT), adrenocorticotropic hormone (ACTH), corticotropin releasing hormone (CRH), norepinephrine (NE), 5-hydroxytryptamine (5-HT) and 5-hydroxyindoleacetic acid (5HIAA) levels are measured to evaluate the function of the hypothalamic-pituitary-ovarian (HPO) and hypothalamic-pituitary-adrenal (HPA) axis. The results showed that OVX-CUMS significantly decrease sucrose preference rate in SPT, increase immobility time in FST and OFT, and decrease movement distance and stand-up times in OFT. HSO treatment significantly improves depression-like behaviors, upregulates the expression of ERα and ERβ, improves HPO axis function by increasing E2 levels and decreasing FSH and LH levels, reverses HPA axis hyperactivation by decreasing CORT, ACTH, and CRH levels, and upregulates NE, 5-HT, and 5HIAA levels in model rats. The findings suggested that HSO could improve depression-like behavior in OVX-CUMS rats by regulating HPO/HPA axis function and neurotransmitter disturbance.

Effects of atorvastatin in suppressing pulmonary vascular remodeling in rats with chronic obstructive pulmonary disease

ObjectiveTo investigate the effects of atorvastatin calcium on pulmonary vascular remodeling, the authors explored the regulatory mechanism of Histone Deacetylation Enzyme-2 (HDAC2) in rats with Chronic Obstructive Pulmonary Disease (COPD), and provided a new direction for drug treatment in the progression of vascular remodeling. MethodsEighteen female SD rats were randomly divided into control (Group S1), COPD (Group S2), and atorvastatin calcium + COPD (Group S3) groups. A COPD rat model was established by passive smoking and intratracheal injection of Lipopolysaccharide (LPS). Haematoxylin and eosin staining and Victoria Blue + Van Gibson staining were used to observe pathological changes in the lung tissue. The pulmonary vascular inflammation score was calculated, and the degree of pulmonary vascular remodeling was evaluated. The ratio of Muscular Arteries in lung tissue (MA%), the ratio of the vessel Wall Area to the vessel total area (WA%), and the ratio of the vessel Wall Thickness to the vascular outer diameter (WT%) were measured using imaging software. The expression of HDAC2 was measured using western blotting, ELISA (Enzyme-Linked Immunosorbent Assay), and qPCR (Real-time PCR). ResultsCompared with the control group, the degree of pulmonary vascular inflammation and pulmonary vascular remodeling increased in rats with COPD. The WT%, WA%, and lung inflammation scores increased significantly; the expression of HDAC2 and HDAC2mRNA in the serum and lung tissue decreased, and the level of Vascular Endothelial Growth Factor (VEGF) in the lung tissues increased (p < 0.05). Compared with the COPD group, the lung tissues from rats in the atorvastatin group had fewer inflammatory cells, and the vascular pathological changes were significantly relieved. The WT%, WA%, and lung inflammation scores decreased significantly; the expression of HDAC2 and HDAC2mRNA in the serum and lung tissues increased, and the level of VEGF in the lung tissues decreased (p < 0.05). ConclusionThe present study revealed that atorvastatin calcium could regulate the contents and expression of HDAC2 in serum and lung tissues and inhibit the production of VEGF, thereby regulating pulmonary vascular remodeling in a rat model with COPD.

Effect of microsampling (50 μL) on toxicological evaluation of methapyrilene, a hepatotoxic substance, in a collaborative 28-day study in female rats.

Although microsampling of blood is recommended to promote the 3Rs in toxicokinetic (TK) evaluation, there are few reports applying microsampling in actual toxicity evaluation. Here, we assessed the effects of microsampling on toxicological evaluation of methapyrilene hydrochloride, a hepatotoxic substance. Female SD rats received methapyrilene hydrochloride orally at dose levels of 0 (vehicle), 10, and 30 mg/kg BW, once daily for 4 weeks. Each dose level included a microsampling group and a non-microsampling group (n = 5). In the microsampling groups, blood sampling (50 µL/time point) was performed at 6 time points on day 1 of administration and 7 time points on day 27-28; all the animals underwent necropsy on day 29. Toxicity studies and TK analysis were performed, and through these studies in 2 organizations, cross-organization validation of the effect on toxicity evaluation was conducted. In one organization, microsampling obscured changes in some parameters in hematology due to the administration of methapyrilene hydrochloride. In the other organization, although the relationship between the developing pattern of histopathological findings in the liver and the blood sampling was suspected, it was associated with poor reproducibility; this was considered as a change within a variation range of biological reactions. Each of these phenomena was observed in only one organization without consistency. In both organizations, no effect of blood microsampling was observed in other endpoints. In conclusion, microsampling is considered to be a technique applicable to safety studies of drugs showing hepatotoxicity, as it did not show a marked influence on the toxicological evaluation of methapyrilene hydrochloride.

Effect of Magnetic Nanoparticles on Hormone Level Changes During Perimenopausal Period and Regulation of Bone Metabolism.

This work aimed to explore the effect of nerve magnetic stimulation based on superparamagnetic Fe3O4 nanoparticle (NP) on bone metabolism during the perimenopausal period. First, the multifunctional water-soluble polymer PTMP-PMAA was utilized as the ligand. PTMP-MAA@ Fe3O4 NP with high magnetization was prepared by the co-precipitation method, and NPX diffraction pattern analysis and in vitro stability analysis were implemented. Then, NPs were co-cultured with 293T cells, and the cytotoxicity was detected by the CCK-8 method. Subsequently, 3-month-old female young SD rats and 11~15-month-old natural menopausal SD rats were taken as the research objects. According to the vaginal smear, the rats were randomly rolled into a young control, perimenopausal period model, estrogen treatment, and osteoporosis prevention groups. Rats in the estrogen treatment group were given Premarin suspension by gavage. Rats in the osteoporosis prevention group were injected stereotaxically with PTMP-MAA@ Fe3O4 NP suspension, and a rotating magnetic field was applied to the brain for nerve magnetic stimulation. The rats were sacrificed three days after treatment and brain tissues were taken for pathological analysis. Rat humerus was weighted and dual-energy X-ray was utilized to determine bone density and bone mineral content. Serum was collected and radioimmunoassay and ELISA were employed to detect estradiol (E2), osteocalcin (Boneglaprotein, BGP), oxytocin (OT), bone alkaline phosphatase (BALP), type I collagen carboxy-terminated cross-linked peptide (CTX-I), and tartrate-resistant acid phosphatase (TRACP-5b) in the serum of rats in each group. The results showed that PTMP-MAA@ Fe3O4 NP had good biocompatibility, and the CCK-8 test results showed that PTMP-MAA@ Fe3O4 NP had low cytotoxicity. Compared with the young control group, the humeral dry weight, wet weight, bone density, and bone mineral content, serum E2, OT, and BGP content in the perimenopausal period model group were reduced, while the serum BALP, CTX-I, and TRACP -5b content was increased (P<0.05). It was verified that nerve magnetic stimulation based on PTMP-MAA@ Fe3O4 NP increased the serum estrogen level of female rats during the perimenopausal period, increased the bone density of rats, promoted bone formation, and regulated bone metabolism.

Relationship between mechanical and microstructural parameters of rat lumbar spine in different ages

Exploring the relationships between microstructure and mechanical properties of bones may provide effective suggestions for increasing bone strength and reducing osteoporotic fracture. In this research, the tissue-level mechanical parameters, microstructure parameters of cancellous bone, and apparent mechanical parameters of L6 vertebral body were calculated in female SD rats aged 1-, 3-, 5-, 7-, 9-, 11-, 13-, 15-, 16-, and 17-month-old. Data were processed with Kruskal–Wallis test, linear regression and Spearman’s rank correlation analysis. Appropriately increasing the plate Tb.N could enhance mechanical properties of bone. Tb.Th and Tb.N were two key factors in determining the tissue-level mechanical properties of cancellous bone. The microstructure could significantly predict mechanical parameters. Our findings may help to further understand the mechanism of osteoporotic fractures.

Hypoxia inducible factor-1α mediates the mechanism of the Hedgehog pathway in tendinopathy repair by Asperosaponin VI.

Our previous study found that asperosaponin VI (ASA VI) has a positive effect on the repair of tendinopathy. However, its molecular biological mechanism is unclear. To investigate the role of hypoxia inducible factor-1α (HIF-1α) in mediating the hedgehog (Hh) pathway in tendinopathy repair by ASA VI. A total of 36 2-month-old female SD rats were classified into the normal group (NG, n=10) and tendinopathy model group (n=26). The tendinopathy model group was further divided into the model group (MG), ASA VI group (AG), and triamcinolone acetonide+lidocaine group (TG). Compared with those in the MG group, IL-1 mRNA was significantly downregulated and IL-4 and IL-10 were increased in the AG group (P<0.01). The mRNA expression levels of MMP3, TIMP3, VEGF-A, KDR, and VWF mRNA decreased (P<0.01). Immunofluorescence staining revealed that CD31/endomucin levels were significantly attenuated. Scx, Mkx, EYA1, EYA2, COL1, COL3, and TNC mRNA levels showed significant differences (P<0.01). Immunofluorescence staining suggested the upregulation of Scx and the downregulation of Sox9. Shh, Ptch1, Smo, Gli1, Cyc-D1, Cyc-E1, and c-Myc mRNA levels were downregulated (P<0.01). The protein expression levels of Gli 1, Shh, and Ptch1 decreased significantly (P<0.01). The immunofluorescence staining levels of Shh, Ptch, and Gli 1 significantly decreased. ASA VI inhibits local vascular hyperproliferation and downregulates the HIF-1α/Hh pathway to promote the tendinous differentiation of tendon stem/progenitor cells and the repair of tendinopathy. The effect of ASA VI on HIF-1α levels may be an effective target in the treatment of tendinopathy.

Effect of olfactory ensheathing glia clone TEG3 on neuroprotection, microglial activation and gene expression profile of allotransplanted retina with and without immunosuppression

AbstractPurpose: Olfactory ensheathing glia (OEG) is a macroglia from the embryonic ectoderm, with unique functions in guiding sensory axons from the olfactory mucosa (PNS) to the olfactory bulb (CNS). These properties, together with their high secretion of trophic factors, have made its use a field of study in axonal regeneration. However, their in vivo administration in the retina had never been tested, so we set out to evaluate their actions in the retina without injury and after an optic nerve crush (ONC) model. In addition, to simulate the clinical conditions of an allogeneic cell transplant, we evaluated how their properties were altered when combined with immunosuppressive treatment.Methods: Wistar TEG3‐GFP+ cells were administered into the vitreous of SD female rats. Experimental groups: transplantation in intact retinas, in retinas just after ONC, and both with systemic immunosuppression. In some animals of each group, the retina was dissected for anatomical analysis and the total number of retinal ganglion cells (RGCs) was counted and the morphological activation of microglia was studied. The retinas of the remaining animals were dissected fresh for the study of gene expression by real‐time quantitative PCR. Animals injected with vehicles after ONC and intact animals without injury were used as controls. n = 3–8 animals/group/analysis.Results: TEG3 neuroprotected RGCs after ONC at 7 days (18% more survival compared to vehicle group p &lt; 0.05) but caused a large activation of microglia cells coupled with a 33% (p &lt; 0.01) reduction of RGCs 21 after transplantation in intact retinas. Immunosuppression had no effect on neuroprotective properties but did recover some of the death observed at 21 days in intact retinas (up to a 17%, p &lt; 0,05). Immunosuppression reduced the expression of pro‐inflammatory molecules (Tnf, Il6), macrogliosis markers (Gfap) and macrophage markers (Ptprc) in favour of anti‐inflammatory ones (Il10). However, markers of pyroptotic cell death (Casp4, Gsdmd) did not change after treatment, which would explain the elevated microglial activation and death observed at 21 days in intact and grafted retinas.Conclusions: TEG3 has a dual effect neurotoxic and neuroprotective in retina.

4D label-free quantitative proteomics analysis to screen potential drug targets of Jiangu Granules treatment for postmenopausal osteoporotic rats.

Background: Postmenopausal osteoporosis (PMOP) is a disease with a high prevalence in postmenopausal women and is characterized by an imbalance in bone metabolism, reduced bone mass, and increased risk of fracture due to estrogen deficiency. Jiangu granules (JG) is a compound prescription used in traditional Chinese medicine to treat PMOP. However, its definitive mechanism in PMOP is unclear. This study used a 4D label-free quantitative proteomics method to explore the potential therapeutic mechanism of JG in an ovariectomy (OVX) rats' model. Materials and methods: A rat model of PMOP was established by removing the ovaries bilaterally. Nine 3-month-old specific-pathogen-free female SD rats. The nine rats were randomly divided into 3 groups (n = 3 in each group): the sham-operated group (J), the ovariectomy group (NC), and the JG treatment (ZY) group. Proteins extracted from the bone tissue of the lumbar spine (L3, L4) of three groups of rats were analyzed by 4D label-free quantitative proteomics, and proteins differentially expressed after JG treatment and proteins differentially expressed after de-ovulation were intersected to identify proteins associated with the mechanism of PMOP by JG treatment. Result: There were 104 up-regulated and 153 down-regulated differentially expressed proteins (DEPs) in the J group vs. NC group, 107 up-regulated and 113 down-regulated DEPs in the J group vs. ZY group, and 15 up-regulated and 32 down-regulated DEPs in the NC group vs. ZY group. Six potential target proteins for JG regulation of osteoblast differentiation in OVX rats were identified by taking intersections of differential proteins in the J group vs. NC group and NC group vs. ZY group. Conclusion: JG may exert therapeutic effects by modulating the expression levels of target proteins associated with osteoblast differentiation to enhance osteoblast differentiation in OVX rats. These results further uncovered the target proteins and specific mechanisms of JG in treating PMOP, providing an experimental basis for the clinical application of JG in treating PMOP.

Intervention effects of estradiol on myocardial ischemia- reperfusion injury of rat and its mechanisms

Objective: To study the effects of estradiol (E2) on alleviating myocardial ischemia/reperfusion(I/R) injury through estrogen receptorβ(ERβ) mediated extracellular regulated protein kinases(ERK) pathway activation. Methods: Eighty-four adult female SD rats were ovariectomized and randomly divided into control group, NC siRNA adeno-associated virus (AAV) group received sham operation, the myocardial I/R injury model was prepared by ligation of the left anterior descending coronary artery in I/R group, E2+I/R group, NC siRNA AAV+I/R group, NC siRNA AAV+E2+I/R group and ERβ-siRNA AAV+E2+I/R group. E2+I/R group, NC siRNA AAV+E2+I/R group and ERβ-siRNA AAV+E2+I/R group were treated with E2 0.8 mg/kg by gavage for 60 days before modeling. NC siRNA AAV+I/R group, NC siRNA AAV+E2+I/R group, and ERβ-siRNA AAV+E2+I/R group were treated with AAV by caudal vein injection 24 h before modeling. After 120 min of reperfusion, the contents of serum lactate dehydrogenase (LDH), phosphocreatine kinase (CK), phosphocreatine kinase isoenzyme (CK-MB), myocardial infarction area and the expressions of ERβ, p-ERK, the contents of tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1 β), malondialdehyde (MDA) and total antioxidant capacity (T-AOC) in myocardium were measured. Results: The contents of serum LDH, CK, CK-MB, myocardial infarction area and the contents of TNF-α, IL-1 β, MDA in myocardium of I/R group were higher than those of the control group, the expression levels of ERβ and p-ERK and the content of T-AOC were lower than those in the control group (P＜0.05). The contents of serum LDH, CK and CK-MB, myocardial infarction area and the contents of TNF-α, IL-1 β and MDA in myocardium of E2+I/R group were lower than those of the I/R group, the expression levels of ERβ and p-ERK and the content of T-AOC were higher than those of the I/R group(P＜0.05). After knockdown ERβ by caudal vein injection of ERβ-siRNA AAV, the contents of serum LDH, CK and CK-MB, myocardial infarction area and the contents of TNF-α, IL-1 β and MDA in myocardium of ERβ-siRNA AAV+E2+I/R group were higher than those of NC-siRNA AAV+E2+I/R, the expression levels of ERβ and p-ERK and the content of T-AOC were lower than those of NC-siRNA AAV+E2+I/R(P＜0.05). Conclusion: E2 has protective effects on myocardial I / R injury in ovariectomized rats, which are related to the promotion of ERβ mediating the activation of ERK pathway, reducing inflammatory and oxidative stress responses.

Jian-Pi-Yi-Shen decoction inhibits mitochondria-dependent granulosa cell apoptosis in a rat model of POF.

As a widely applied traditional Chinese medicine (TCM), Jian-Pi-Yi-Shen (JPYS) decoction maybe applied in curing premature ovarian failure (POF) besides chronic kidney disease (CKD). In vivo experiments, 40 female SD (8-week-old) rats were randomized into four groups, namely, control group (negative control), POF model group, JPYS treatment group, and triptorelin treatment group (positive control). JPYS group was treated with JPYS decoction (oral, 11 g/kg) for 60 days, and the triptorelin group was treated with triptorelin (injection, 1.5 mg/kg) for 10 days before the administration of cyclophosphamide (CTX) (50 mg/kg body weight) to establish POF model. We examined apoptosis, mitochondrial function, and target gene (ASK1/JNK pathway and mitochondrial fusion/fission) expression. In vitro experiments, the KGN human granulosa cell line was used. Cells were pretreated with CTX (20, 40, and 60 μg/mL) for 24 h, followed by JPYS-containing serum (2, 4, and 8 %) for 24 h. Thereafter, these cells were employed to assess apoptosis, mitochondrial function, and target gene levels of protein and mRNA. In vivo, JPYS alleviated injury and suppressed apoptosis in POF rats. In addition, JPYS improved ovarian function. JPYS inhibit apoptosis of granulosa cells through improving mitochondrial function by activating ASK1/JNK pathway. In vitro, JPYS inhibited KGN cell apoptosis through inhibited ASK1/JNK pathway and improved mitochondrial function. The effects of GS-49977 were similar to those of JPYS. During POF, mitochondrial dysfunction occurs in the ovary and leads to granulosa cell apoptosis. JPYS decoction improves mitochondrial function and alleviates apoptosis through ASK1/JNK pathway.

Long-term environmental exposure of darkness induces hyperandrogenism in PCOS via melatonin receptor 1A and aromatase reduction.

Polycystic ovary syndrome (PCOS) is a common and complex disorder impairing female fertility, yet its etiology remains elusive. It is reported that circadian rhythm disruption might play a crucial role in PCOS pathologic progression. Here, in this research, we investigated the effect of environmental long-term circadian rhythm dysfunction and clarified its pathogenic mechanism in the development of PCOS, which might provide the targeted clinical strategies to patients with PCOS. Female SD rats were used to construct a circadian rhythm misalignment model with constant darkness (12/12-h dark/dark cycle), and the control group was kept under normal circadian rhythm exposure (12/12-h light/dark cycle) for 8 weeks. We measured their reproductive, endocrinal, and metabolic profiles at different zeitgeber times (ZTs). Different rescue methods, including melatonin receptor agonist and normal circadian rhythm restoration, and in vitro experiments on the KGN cell line were performed. We found that long-term darkness caused PCOS-like reproductive abnormalities, including estrous cycle disorder, polycystic ovaries, LH elevation, hyperandrogenism, and glucose intolerance. In addition, the expression of melatonin receptor 1A (Mtnr1a) in ovarian granulosa cells significantly decreased in the darkness group. Normal light/dark cycle and melatonin receptor agonist application relieved hyperandrogenism of darkness-treated rats. In vitro experiments demonstrated that decreased MTNR1A inhibited androgen receptor (AR) and CYP19A1 expression, and AR acted as an essential downstream factor of MTNR1A in modulating aromatase abundance. Overall, our finding demonstrates the significant influence of circadian rhythms on PCOS occurrence, suggests that MTNR1A and AR play vital roles in pathological progression of hyperandrogenism, and broadens current treatment strategies for PCOS in clinical practice.

Aloe polysaccharide promotes osteogenesis potential of adipose-derived stromal cells via BMP-2/Smads and prevents ovariectomized-induced osteoporosis

BackgroundAloe polysaccharide (AP) is a type of an active macromolecule of Aloe vera, which contributes to its function. However, whether AP possesses anti-osteoporosis properties is unknown.MethodsAdipose-derived stromal cells were treated with different concentrations of AP. Early and late osteogenesis were, respectively, evaluated by ALP and Alizarin Red S staining. The effect of AP on the processes of adipogenesis inhibition in ADSCs was analyzed by oil red O staining. Western blot was used to assess the expression of osteogenic and adipogenic related factors. Then, Noggin was administered to further confirm the mechanism by which AP promotes the osteogenesis of ADSCs. Finally, 40 female SD rats were classified into a bilateral laparotomy group (Sham group) and three bilateral ovariectomy groups: OVX group, OVX + AP group, and OVX + AP + Noggin group. The bilateral rat femurs were collected to perform micro-CT scanning, HE, Masson trichrome, and Oil red O staining.ResultsThe results indicated that AP could increase ALP expression and calcium deposition. Through molecular mechanisms, AP promotes the protein expression of COL1A1, OPN, and ALP in ADSCs, but downregulates the expression of PPARγ. Also, AP directs ADSCs’ fate by stimulating the BMP2/Smads signaling pathway. In vivo, the rat AP-treated had more trabecular bone than the OVX rat, indicating partial protection from cancellous bone loss after treatment with AP.ConclusionOur results show that AP may promote osteogenesis of ADSCs through BMP-2/Smads signaling pathway and inhibits lipogenic differentiation. Thus, AP might be a promising alternative medicine to treat postmenopausal osteoporosis.

Gender-related differences in the effects of Inonotus obliquus polysaccharide on intestinal microorganisms in SD rats model.

Natural edible fungal polysaccharides are of research and application value for the prevention of diseases by improving the microenvironment within the intestine. Inonotus obliquus polysaccharide (IOP) extracts have strong antioxidant, anti-inflammatory, and other biological activities, and as such, it could be used as prebiotics to improve the viability of intestinal microbes, maintain intestinal homeostasis and improve intestinal immunity. The effects of sex on intestinal microbiota after IOP absorption was determined. In this study, IOP had different effects on the intestinal flora of male and female rats, with the diversity and richness showing opposite changes. At the same time, after IOP intervention, changes in the dominant intestinal flora of female rats was less compared with that of males. In addition, while Clostridia, Lactobacillus and Roseburia were the dominant intestinal microbes in female rats, males had mainly Bacteroidota from different families and genera, along with an increasing proportion of Muribaculaceae from different families and genera. IOP could further regulate the intestinal microenvironment of male and female SD rats by enhancing the vitality of their dominant microorganisms, and for both sexes, this enabled the screening of dominant microflora that were conducive to the balance of the intestinal flora. These results help to understand the effects of sex-related differences on the composition of the intestinal microbiota as well as on diseases.

Improvement effect of Lycium barbarum polysaccharide on the intestinal flora of pregnant rats and their offspring under chronic stress

Objective: To understand the improvement effect of Lycium barbarum polysaccharide (LBP) on the intestinal flora of mother mice during pregnancy and their offspring who experienced chronic stress, and provide new ideas for improving the effect of stress on the intestinal tract. Methods: From July to October 2019, 24 SPF-grade female SD rats were selected and divided into control group, stress group, and stress+LBP group, with 8 rats in each group. A chronic unpredictable mild stimulation model during pregnancy was established (21 days) , and 40 mg/kg LBP solution was administered by gavage on the 8th day of stress. Venous blood from the medial canthus of the female mice was collected on the 1st day before stress and on the 1st, 7th, 14th and 21st days, respectively. Cortisol was measured and corticosterone concentration was calculated. The fresh feces of famale mice after stress and 20-day postnatal offspring mice were collected, and Illumina Miseq sequencing technology, alpha diversity and community composition were used to analyze the diversity and structure of intestinal flora. Results: On the 7th and 14th days of stress, the plasma corticosterone concentration of female mice in the stress group and stress+LBP group was higher than that in the control group (P<0.05) . In the Alpha diversity of female mice, the Ace index of the stress group was lower than that of the control group (P<0.05) . The analysis of intestinal flora structure showed that at the species level, the proportions of Lachnospiraceae and Lactobacillus in the stress+LBP group were higher than those in the stress group and control group. At the order level, the proportion of Clostridiales in the stress+LBP group was higher than that in the stress group and lower than that in the control group, while the proportion of Lactobacillales was higher than that in the stress group and control group. In the Alpha diversity of the offspring group, the Shannon index, Ace index and Chao index of the stress+LBP offspring group were higher than those of the stress offspring group (P<0.05) . The proportion of Lactobacillus in the stress+LBP offspring group was higher than that in the control offspring group and stress offspring group, and the proportions of Lachnospiraceae and Ruminococcaceae in the stress+LBP offspring group were higher than those in the stress offspring group, the proportion of Bacteroidales in the stress+LBP offspring group was lower than that in the stress offspring group, and the proportion of Clostridiales in the stress+LBP offspring group was higher than that in the stress and control offspring groups. Conclusion: The intervention of LBP may improve the changes in the intestinal flora diversity, abundance and flora structure of mother mice and offspring caused by pregnancy stress, thereby maintaining the balance of intestinal flora.

Simultaneous determination and pharmacokinetic study of five compounds from total extract of Clinopodium chinense in abnormal uterine bleeding rat plasma by UPLC-MS/MS

Clinopodium chinense, a traditional folk medicinal herb, has been used to treat abnormal uterine bleeding(AUB) for many years. Saponins and flavonoids are the main active components in C. chinense. To study the pharmacokine-tics of multiple components from the total extract of C. chinense(TEC), we established a sensitive and rapid method of ultra-perfor-mance liquid chromatography coupled with tandem mass spectrometry(UPLC-MS/MS) for simultaneous determination of five compounds in the plasma of AUB rats. After validation, the AUB model was established with SD female rats which got pregnant on the same day by gavage with mifepristone(12.4 mg·kg~(-1)) and misoprostol(130 μg·kg~(-1)). The established method was applied to the detection of hesperidin, naringenin, apigenin, saikosaponin a, and buddlejasaponin Ⅳb in AUB rats after the administration of TEC. The pharmacokinetic parameters were calculated by DAS 2.0. The five compounds showed good linear relationship within the detection range. The specificity, accuracy, precision, recovery, matrix effect, and stability of the method all matched the requirements of biolo-gical sample detection. The above 5 compounds were detected in the plasma of AUB rats after the administration of TEC. The C_(max) va-lues of hesperidin, naringenin, apigenin, saikosaponin a, and clinoposide A were 701.6, 429.5, 860.7, 75.1, and 304.1 ng·mL~(-1), respectively. All the compounds owned short half-life and quick elimination rate in vivo, and the large apparent volume of distribution indicated that they were widely distributed in tissues. Being rapid, accurate, and sensitive, this method is suitable for the pharmacokinetic study of extracts of Chinese herbal medicines and provides a reference for the study of pharmacodynamic material basis of C. chinense in treating AUB.

Protective Effect of Rho-Associated Protein Kinase (ROCK) Activated by Bone Marrow Mesenchymal Stem Cells on Bone Strength and Osteoblasts in Rats with Osteoporosis

This study assesses BMSCs’ effect on bone strength and osteoblast activity in rats with osteoporosis. 40 SD female rats were assigned into normal (A) group, model (B) group, puerarin (C) group, and BMSCs (D) group followed by analysis of bone strength by small animal bone strength analyzer and the expression of ROCK1 protein in tibia by immunohistochemistry. The bone strength of group B was lower than group A (P &lt; 0.05), and higher in groups C and D than group B (P &lt; 0.05) with further higher in group D than group C (P &lt; 0.05). Group A showed obvious mesh cancellous bone trabecular bone and continuity, compact structure, and group B cells appeared loose and irregular distribution, parts cancellous bone trabeculae decrease and part of the fracture, and arranged in sparse irregular distribution. The decrease of trabecular bone in group C and D is not more obvious than group B and the morphology in group D got better improved than in group C. Group B showed lower proliferation of osteoblasts and ALP activity than group A (P &lt; 0.05) and the proliferation and ALP activity in groups C and D was increased significantly compared with group B (P &lt; 0.05). ROCK1 expression was significantly lower in group B than group A (P &lt; 0.05) higher in groups C and D than group B (P &lt; 0.05). Bone marrow mesenchymal stem cells (BMSCs) can effectively increase the bone strength and activity of osteoblasts in rats with osteoporosis, and promote the activation of ROCK signal.

Sexual dysfunction in university students: prevalence and association with quality of life

Background: Female sexual dysfunctions impacts QoL, affecting personal relationships and decreasing self-esteem, self-confidence and libido. Aims: To establish the prevalence of SD in university students of the Physiotherapy course at ULBRA Canoas/RS and to verify the association with QoL. Method: Cross-sectional study, through questionnaires on the Google Forms, through the FSFI (Female Sexual Function Index) and the WHOQOL-bref. Results: 69 students were included, mean age 25.5 years. Total prevalence of SD was 36%, with pain and lubrication being the most affected domains (average of 4.9). QoL was very good, however, when comparing the risk of SD with the WHOQOL-bref domains, it was observed that the psychological, social relationship and general quality of life domains had the lowest scores in the WHOQOL-bref. high risk group for DS. The domains of satisfaction and general quality of life showed a statistically significant and moderate positive correlation (r=0.40). Conclusion: Female SD is prevalent in young university students and has a negative impact on some aspects of QoL.

Effects of ferric ammonium citrate on iron accumulation, bone turnover and bone density in ovariectomized rat models with osteoporosis.

To analyze the effect of ferric ammonium citrate on iron accumulation, bone turnover and bone density in ovariectomized rat models with osteoporosis, 40 female SD rats were randomized into four groups, that were, sham-operated, model, low and high-dose ferric ammonium citrate groups (i.e. low and high-dose groups) respectively, each of them had ten rats. Except for the sham-operated group, bilateral ovariectomy was performed in the other groups to establish models with osteoporosis; one week after the operation, those in the low and high-dose groups were given 90 mg/kg and 180 mg/kg ferric ammonium citrate, respectively. Those in the other two groups received isodose saline for nine weeks, with the frequency of twice per week. The changes in bone tissue morphology, serum ferritin concentration, tibial iron content, serum osteocalcin, carboxyl terminal peptide (β - CTX), bone density, bone volume fraction and trabecular thickness were compared. Results showed that the rats in the low and high-dose groups contained a higher concentration of serum ferritin and tibial iron content compared to the other groups (P < 0.05). In contrast to the model group, the bone trabeculae in the low and high-dose groups were sparse in morphology and increased in spacing. It was obvious that the rats contained more osteocalcin and β - CTX in the model group, the low and high-dose groups versus the sham-operated group (P < 0.05), and those had more β - CTX in the high-dose group versus the model group and the low-dose group (P < 0.05). The bone density, bone volume fraction and trabecular thickness of the rats in the model group, the low and high-dose groups showed lower versus the sham-operated group (P < 0.05); those in the low and high-dose groups significantly presented lower bone density and bone volume fraction versus the model group (P < 0.05). Iron accumulation can aggravate osteoporosis in ovariectomized rats, and its mechanism may be associated with accelerating bone turnover, promoting bone absorption, reducing bone density and sparsely trabecular structure. Therefore, it is particularly important to understand iron accumulation in postmenopausal osteoporosis patients.