Role and mechanism of action of phytoestrogen biochanin A in improving liver fibrosis in ovariectomized mice / 临床肝胆病杂志

ObjectiveTo investigate the effect of phytoestrogen biochanin A （BCA） on liver fibrosis induced by CCl4 in female mice with bilateral oophorectomy （ovariectomized） and its mechanism. MethodsA total of 50 ovariectomized Kunming mice were selected and given intraperitoneal injection of CCl4 to establish a model of liver fibrosis， and then according to body weight， they were randomly divided into model group， positive control group， and low-， middle-， and high-dose BCA groups， with 10 mice in each group. In addition， 10 female mice in the same litter were given resection of a small amount of adipose tissue near both ovaries to establish the sham-operation group. The mice in the positive control group were given estradiol 2 mg/kg by gavage， and those in the low-， middle-， and high-dose BCA groups were given BCA by gavage at a dose of 25， 50， and 100 mg/kg， respectively， once a day for 7 consecutive weeks； the mice in the sham-operation group and the model group were given an equal volume of 0.5% sodium carboxymethyl cellulose solution by gavage. The mice were anesthetized and sacrificed after administration to collect samples. Liver index and uterus index were measured； HE staining and Masson staining were used to observe liver histopathological changes； the biochemical analysis was used to measure the activity of aspartate aminotransferase （AST） and alanine aminotransferase （ALT）； ELISA was used to measure the levels of interleukin-6 （IL-6） and tumor necrosis factor-α （TNF-α） in liver tissue， and Western blot was used to measure the relative protein expression levels of collagen Ⅰ， transforming growth factor-beta 1 （TGF-β1）， alpha-smooth muscle actin （α-SMA）， estrogen receptor beta （ERβ）， and p-NF-κBp65/NF-κBp65 in liver tissue. The t-test was used for comparison of continuous data between two groups； a one-way analysis of various was used for comparison between multiple groups， and the least significant difference t-test was used for further comparison between two groups. The Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between multiple groups and further comparison between two groups. ResultsCompared with the sham-operated group， the model group had a significant increase in liver index and a significant reduction in uterus index， as well as significant increases in the activities of serum AST and ALT， the levels of IL-6 and TNF-α in liver tissue， and the protein expression levels of collagen Ⅰ， TGF-β1， α-SMA， and p-NF-κBp65/NF-κBp65 in liver tissue （all P<0.05）， with no significant change in the expression of ERβ in liver tissue （P>0.05）， and the model group showed significant fibrosis lesions in the liver， such as hepatocyte edema， steatosis， and necrosis with inflammatory cell infiltration and hyperplasia， deposition， and staggered distribution of collagen fibers. Compared with the model group， the low-， middle-， and high-dose BCA groups had significant reductions in liver index， the activities of serum AST and ALT， the levels of IL-6 and TNF-α， and the protein expression levels of collagen Ⅰ， TGF-β1， α-SMA， and p-NF-κBp65/NF-κBp65 in liver tissue （all P<0.05）， with no significant change in uterine index （P>0.05）， as well as a significant increase in the protein expression level of ERβ in liver tissue （P<0.05） and varying degrees of improvement in liver fibrosis lesions. ConclusionBCA can effectively improve CCL4-induced liver fibrosis in ovariectomized female mice， possibly by upregulating ERβ to inhibit the NF-κB signaling pathway and then alleviating inflammatory response.

Lycopene enhances bone neoformation in calvaria bone defects of ovariectomized rats

Abstract Osteoporosis can affect a significant part of the population and fractures are the most common complications associated with this disease, leading to high public health costs. Thus, the prevention of fractures is relevant to individuals with signs and symptoms as well as to the health system. Postmenopausal osteoporosis has been associated with oxidative stress, emphasizing the importance of an efficient defense system to maintain bone health. Lycopene is a carotenoid with antioxidant properties that may stimulate osteoblastogenesis and inhibit osteoclastogenesis. The purpose of this investigation was to analyze the influence of lycopene in the bone neoformation of calvaria defects in ovariectomized rats utilizing the concentration of 45 mg/kg. Wistar Hannover female rats were divided into ovariectomized and sham groups. The ovariectomized animals received 45 mg/kg lycopene (OvxL) or water (Ovx) by daily gavage the day after ovariectomy/sham surgery for 16 weeks. Twelve weeks after ovariectomy, there were performed 5-mm calvaria defects followed by euthanasia after 4 weeks. Samples of bone tissue were collected to perform morphological and morphometrical analysis of the neoformed bone area, and percentage with Software Image J. Morphological evaluation showed mature bone with more osteocytes in the group OVxL when compared to the other groups. The morphometrical analysis demonstrated a significant increase of bone neoformation in the group OvxL (p<0.05). The data obtained suggest that lycopene benefits bone repair in the absence of estrogenic hormones.

Resumo A osteoporose afeta grande parte da população e as fraturas são as complicações mais importantes relacionadas a essa doença, gerando altos gastos para o poder público. Dessa forma, a prevenção de fraturas decorrentes da osteoporose torna-se relevante tendo em vista que gera benefícios tanto para o indivíduo acometido pela doença quanto para o sistema de saúde. A osteoporose pós menoupasa tem sido associada ao estresse oxidativo, portanto, um eficiente sistema de defesa antioxidante é primordial para a manutenção da saúde óssea. O licopeno é um carotenoide antioxidante que aparentemente estimula a osteoblastogênese e inibe a osteoclastogênese. O objetivo deste estudo foi analisar a influência do licopeno na neoformação óssea em defeitos de calvária em ratas ovariectomizadas utilizando a concentração de 45 mg/kg. Foram utilizados 15 ratas Wistar Hannover pesando aproximadamente 200g, sendo que 10 animais foram submetidos à ovariectomia bilateral e 5 (Grupo Sham) foram submetidos à simulação da cirurgia de ovariectomia bilateral. Os animais ovariectomizados foram divididos aleatoriamente em 2 grupos: Ovariectomizado (Ovx) e Ovariectomizado Licopeno (OvxL) que receberam água e licopeno respectivamente, por sonda gástrica, diariamente. As administrações iniciaram-se no dia seguinte à cirurgia de ovariectomia e/ou da exposição dos ovários e foram mantidas por 120 dias, data de realização da eutanásia. O grupo Sham recebeu água diariamente. Noventa dias após a ovariectomia bilateral foram confecionados defeitos ósseos nas calvárias de todos os animais e após trinta dias as ratas foram eutanasiadas. As amostras de tecido ósseo foram coletadas e foi realizado o processamento para a obtenção das lâminas histológicas. Foram realizadas as análises morfológicas e morfométrica, onde foi estimada a área (mm2) e porcentagem (%) relativa de osso neoformado utilizando o Software Image J. A avaliação morfológica evidenciou a ação benéfica do licopeno pois os animais que receberam esse antioxidante apresentaram um tecido ósseo mais maduro, com maior presença de osteócitos quando comparados aos demais grupos. Por meio das análises morfométricas verificou-se maior neoformação óssea para os animais que receberam o licopeno (p<0,05). Diante dos resultados obtidos, concluiu-se que o licopeno na concentração de 45 mg/Kg teve efeito benéfico no processo de reparação, promovendo significante formação óssea frente à ausência de hormônios estrogênicos.

Effect of strength training versus raloxifene on bone weight, blood glucose, lipid and antioxidant profile in ovariectomized rats

We compared the effect of the treatment with strength training (ST) and raloxifene (RALOX) on bone weight, blood glucose, lipid, and antioxidant profile in ovariectomized rats. Twenty-four Wistar rats were distributed into four groups: ovariectomy + VEHICLE (control); ovariectomy + RALOX; ovariectomy + ST; ovariectomy + RALOX + ST. Thirty days after ovariectomy, the animals underwent the treatment with RALOX (750 µcg day-1) and/or ST (three sessions week-1). Thirty days after, all groups were scarified, tibia and femur were weighed, and the blood was collected for analysis of the lipid profile, glucose, and antioxidants catalase (CAT) and glutathione (GSH). The ST group showed greater femur weight (0.82 ± 0.18 g) and RALOX + ST had greater tibia weight (0.61± 0.17 g) than CONTROL with femur weight of 0.65 ± 0.08 g and tibia of 0.49 ± 0.08 g with no differences between treatments (p > 0.05). ST group showed significantly higher catalase (181.7 ± 15.4 µM g-1) compared to the other groups. In contrast, the GSH value was lower in ST group (89.2 ± 8.1 µM g-1) compared to RALOX (175.9 ± 17.1 µM g-1) and RALOX + ST (162.8 ± 12.1 µM g-1), but the values of these two groups did not differ from CONTROL(115.3 ± 21.1 µM g-1). Total cholesterol did not differ between groups (p > 0.05), but exercise alone(54.3 ± 2.5 mg dL-1) or with RALOX (53.0 ± 1.5 mg dL-1) resulted in higher HDL cholesterol than CONTROL (45.5 ± 2.5 mg dL-1). Only RALOX+ST presented lower glucose (140.3 ± 9.7 mg dL-1) values than CONTROL (201.7 ± 30.6 mg dL-1). In conclusion, ST promotes similar benefits on bone and metabolic parameters compared to pharmacological treatment in ovariectomized rats.(AU)

Effects of estrogen and raloxifene on synaptic density in the hippocampal CA1 region of ovariectomized rats

Abstract Introduction The CA1 region of the hippocampus has an important role in learning and memory. It has been shown that estrogen deficiency may reduce the synaptic density in the region and that hormone replacement therapy may attenuate the reduction. Objectives This study aimed to evaluate the effects of estrogen and raloxifene on the synaptic density profile in the CA1 region of the hippocampus in ovariectomized rats. Methods Sixty ovariectomized three-month-old virgin rats were randomized into six groups (n = 10). Treatments started either three days (early treatment) or sixty days (late treatment) after ovariectomy. The groups received propylene glycol vehicle (0.5 mL/animal/day), equine conjugated estrogens (50 μg/animal/day), or raloxifene (3 mg/kg/day) either early or late after ovariectomy. The drugs were administered orally by gavage for 30 days. At the end of the treatments, the animals were anesthetized and transcardially perfused with ether and saline solution. The brains were removed and prepared for analysis under transmission electron microscopy and later fixed. Results Results showed a significant increase in the synaptic density profile of the hippocampal CA1 region in both the early estrogen (0.534 ± 0.026 µ/m2) and the early raloxifene (0.437 ± 0.012 µ/m2) treatment groups compared to the early or late vehicle-treated control groups (0.338 ± 0.038 µ/m2 and 0.277 ± 0.015 µ/m2 respectively). Conclusions The present data suggest that the raloxifene effect may be lower than that of estrogen, even early or late treatment, on synaptic density in the hippocampus.

Ovarian hormones influence immune response to liver ischemia-reperfusion

Liver injury occurs after ischemia and reperfusion (IR), as seen in transplant settings. Sex hormones have been implicated in many pathophysiological mechanisms in females and this could lead to liver protection under inflammatory reperfusion conditions where an excessive immune response occurs. Despite such assumptions, this fact needs to be further investigated. To address this, female and male C57BL/6J mice (8-12 weeks old) were studied. Bilateral ovariectomy (OVX) was performed in females to decrease estradiol levels. IR was performed, and after two weeks, all animals underwent a sham control operation or IR with euthanasia at the following time points after reperfusion: 6, 12, 24, and 48 h. IR triggered an inflammatory process in the liver with recruitment of neutrophils into the parenchyma of male mice. The female sham mice were protected against liver IR presenting no alteration of aminotransferase (ALT) levels compared to males. OVX caused loss of protection, increasing hepatic injury as represented by increased ALT levels and myeloperoxidase (MPO) activity. Female sham mice showed increased Akt phosphorylation and activation, while males showed reduced Akt activation. Estradiol pretreatment recovered ALT levels after IR injury, which was associated with decreased liver injury.

Paeoniflorin ameliorates diabetic cognitive dysfunction by inhibiting TLR4/NF-κB signaling in ovariectomized mice / 中华内分泌代谢杂志

Objective:To investigate the effect of paeoniflorin on toll-like receptor 4(TLR4)/nuclear transcription factor(NF-κB) signaling pathway of streptozotocin combined with ovariectomized mice, and to explore whether it can improve the cognitive impairment of ovariectomized diabetic mice.Methods:Ninety female C57BL/6J mice were divided into SHAM group, ovariectomy group, diabetes group(intraperitoneal injection of STZ 50 mg·kg -1·d -1 for 5 consecutive days), dual model group(DM modeling and OVX operation), paeoniflorin low-dose intervention group(OVX+ STZ+ L-PF 50 mg·kg -1·d -1), paeoniflorin high-dose intervention group(OVX+ STZ+ H-PF 100 mg·kg -1·d -1; all groups n=15). After 8 weeks of paeoniflorin intervention, their cognitive function was tested by behavioral experiments(Morris water maze and Y maze). The pathological changes of hippocampal tissue were observed by HE and Nissl staining. The mRNA expressions of TLR4, tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β) and interleukin-6(IL-6) in hippocampal tissues were detected by real-time fluorescence quantitative PCR. The expression of TLR4, NF-κB P65, TNF-α, IL-6, IL-1β, β-amyloid protein(Aβ), tau proteins, and p-tau proteins were detected by Western blot. Results:Compared with SHAM group, the learning and memory ability of ovariectomy group, diabetes group and dual model group decreased, hippocampal cells were damaged, and the expression of related gene mRNA and protein were increased, especially in dual model group; Compared with dual model group, paeoniflorin intervention could delayed the learning and memory impairment, improve cognitive function, reduce the degree of hippocampal injury, and decrease the expression levels of related gene mRNA and protein, The above changes were the most pronounced at paeoniflorin high-dose intervention group.Conclusion:Paeoniflorin improves cognitive dysfunction in ovariectomized diabetic mice by inhibiting TLR4/NF-κB signaling pathway.

Ovariectomy exacerbates glycerol-induced acute kidney injury in rats / A ovariectomia intensifica a lesão renal aguda induzida pelo glicerol em ratas

Objective: This study aimed to evaluate the effects of ovariectomy on glycerol-induced renal changes in rats. Methods: Twenty-four female Wistar rats were submitted to ovariectomized (OVX) or sham surgery. One week after surgery, the animals received an intramuscular injection (8ml/kg) of 50% glycerol or saline (0.15 M) solution. These animals were divided into the following groups (n=6 per group): Sham, sham-operated female rats injected with saline; OVX, ovariectomized female rats injected with saline; Sham+Gly, sham-operated female rats injected with glycerol; OVX+Gly, ovariectomized female rats injected with glycerol. All rats were euthanized 3 days after the injections and the kidneys were removed for histological and immunohistochemical studies. Blood and urine samples were also collected for renal function studies. Results: The OVX+Gly group presented higher creatinine serum levels, as well as greater fractional excretion of sodium and urinary flow than the Sham+Gly group. Histological lesions and tubulointerstitial staining for macrophages, nuclear factor-kappa B, and nitrotyrosine were more pronounced in the renal cortex of the OVX+Gly group compared to the Sham+Gly group. Conclusion: We conclude that ovariectomy aggravated changes in renal function and structure in glycerol-induced acute kidney injury by the intensification of the proinflammatory tissue response.

Objetivo: Avaliar os efeitos da ovariectomia nas alterações renais induzidas pelo glicerol em ratas. Métodos: Vinte e quatro ratas Wistar foram submetidas à ovariectomia (OVX) ou cirurgia sham (intervenção falsa). Uma semana após a cirurgia, os animais receberam injeção intramuscular (8ml/kg) de glicerol a 50% ou solução salina (0,15 M). As ratas foram divididas nos seguintes grupos (n=6 por grupo): Sham, fêmeas sham-operadas e injetadas com solução salina; OVX, fêmeas ovariectomizadas e injetadas com solução salina; Sham+Gly, fêmeas sham-operadas e injetados com glicerol; OVX+Gly, fêmeas ovariectomizadas e injetadas com glicerol. Todas as ratas foram eutanasiadas 3 dias após as injeções e os rins foram removidos para estudos histológicos e imuno-histoquímicos. Amostras de sangue e urina também foram coletadas para estudos de função renal. Resultados: O grupo OVX+Gly apresentou maiores níveis séricos de creatinina, assim como maiores fração de excreção de sódio e fluxo urinário do que o grupo Sham+Gly. As lesões histológicas e imunomarcação tubulointersticial para macrófagos, fator nuclear-kappa B e nitrotirosina foram mais pronunciadas no córtex renal do grupo OVX+Gly em comparação ao grupo Sham+Gly. Conclusão: Concluímos que a ovariectomia agravou as alterações na função e estrutura renal, na lesão renal aguda induzida por glicerol, pela intensificação da resposta tecidual pró-inflamatória.

Influencia de diferentes materiales de relleno en alvéolos post exodoncia de ratas ovariectomizadas (OVX) / Influence of different graft materials on post extraction sockets of ovariectomized rats (OVX)

RESUMEN: El objetivo del presente estudio fue establecer la influencia de diferentes materiales en el proceso de regeneración ósea de alveolos post exodoncia de ratas hembra adultas ovariectomizadas (OVX). Para ello, se utilizaron 40 ratas sprague dawley, divididas en grupo experimental (OVX) (n=20) y grupo control (Sin ovariectomía) (n=20). Todas las ratas del grupo experimental fueron sometidas a ovariectomía bilateral para simular un estado de osteoporosis inducida por déficit de estrógeno. Posterior a 12 semanas post OVX, las ratas de ambos grupos fueron divididas en 4 subgrupos, en los cuales fue extraído el primer molar superior derecho de cada rata. Posteriormente, las terapias realizadas en los alveolos post-exodoncia fueron: A: (N=5) Alveolo no rellenado para ser utilizado como control negativo. B: (N=5) Aplicación de injerto bifásico (HA+BTCP). C: (N=5) Aplicación de PRF. D: Aplicación de una combinación de injerto bifásico + PRF. Luego de tres semanas se realizó la eutanasia de los animales y obtención de las muestras para los análisis respectivos. Todos los animales sobrevivieron al final del estudio sin ninguna complicación postoperatoria. Los resultados cuantitativos del área ósea interradicular del segundo molar superior, mostraron diferencias estadísticamente significativas entre grupo control y grupo OVX. Mientras que no se observaron diferencias en la descripción histológica ni en el análisis cuantitativo de fibras colágenas tipo I y III. Es posible concluir que el modelo de osteoporosis inducida por déficit de estrógeno modificaría también la microarquitectura ósea de la Maxila. No obstante, nuevos estudios son necesarios para continuar con el estudio de biomateriales para regeneración ósea en modelos de osteoporosis inducida.

SUMMARY: The aim of the present study was to establish the influence of different materials on the process of bone regeneration in post-extraction sockets of ovariectomized (OVX) adult female rats. For this, 40 Sprague Dawley rats were used, divided into an experimental group (OVX) (n=20) and a control group (without ovariectomy) (n=20). All rats in the experimental group underwent bilateral ovariectomy to simulate a state of estrogen deficiency osteoporosis. After 12 weeks post OVX, rats from both groups were divided into 4 subgroups, in which the upper right first molar of each rat was extracted. Subsequently, the therapies performed in the post-extraction sockets were A: (N=5) Unfilled alveolus to be used as a negative control. B: (N=5) Biphasic graft application (HA+BTCP). C: (N=5) PRF application. D: Application of a combination of biphasic graft + PRF. After three weeks, the animals were euthanized, and the samples were obtained for the respective analyses. All animals survived to the end of the study without any postoperative complications. The quantitative results of the interradicular bone area of ??the upper second molar showed significant differences between the control group and the OVX group. While no differences were observed in the histological description or in the quantitative analysis of collagen fibers type I and III. It is possible to conclude that the model of osteoporosis induced by estrogen deficiency would modify the bone microarchitecture of the Maxilla. However, new studies are necessary to continue with the study of biomaterials for bone regeneration in models of induced osteoporosis.

Protective effect of overexpression of thioredoxin reductase 1 on hippocampal cells of ovariectomized rats / 中华行为医学与脑科学杂志

Objective:To investigate the effects of thioredoxin reductase 1(TR1) overexpression on hippocampus in ovariectomized SD rats.Methods:Totally 54 female SD rats were divided into normal group, ovariectomized group and ovariectomized over-expressioned TR1 group (ovariectomy-TR1 group) according to the random number table method with 18 in each group. The overexpressed TR1 vector was constructed by lentivirus, and the recombinant lentivirus was injected into the hippocampus by a brain stereotactic instrument.The mRNA levels and protein levels of TR1, Bcl-2, p53 and p21 in the hippocampus of SD rats were detected by qRT-PCR and Western blot.The expression of Caspase-3 in the hippocampus of SD rats was detected by Western blot. The activity of SOD was measured by the WST-1 cell proliferation and cytotoxicity method, the content of GSH was measured by the microplate method, and the content of MDA in the hippocampus of SD rats was measured by the TBA method. The behavioral changes of SD rats were detected by the open field test and water maze test. GraphPad Prism 9.0 was used for statistical analysis.One-way analysis of variance was used for comparisons among the three groups, and the LSD test was used for further pairwise comparisons, the t-test was used to compare the mean number of samples between the two groups. Results:(1) The mRNA and protein levels of TR1 in hippocampus of ovariectomized rats were lower than those of normal rats ( t=3.125, 4.402, both P<0.05). The mRNA and protein levels of TR1 in hippocampus of ovariectomized-TR1 rats were higher than those of ovariectomized rats ( t=4.945, 4.845, both P<0.05). (2) There were significant differences among the three groups in the escape latency in water maze test, the movement distance and the stay time in central area in the open field test ( F=44.73, 33.67, 6.51, all P<0.05), the movement distance in the open field test of ovariectomized rats was more than that of the normal group ((4 700±141) mm, (3 967±163) mm, P<0.05), the stay time in the central area was longer than that of the normal group ((87.33±3.93) s, (80.83±2.48) s, P<0.05), the movement distance ((4 267±150) mm) and the stay time in the central area ((82.17±3.43) s) of ovariectomized-TR1 group were lower than that of ovariectomized group ( P<0.05). In the water maze test, the escape latency of ovariectomized rats was longer than that of the normal group ((28.67±2.50) s, (19.50±2.59) s, P<0.05), and the escape latency in the ovariectomy-TR1 group((25.00±1.67) s) was shorter than that of ovariectomized TR1 group ( P<0.05). (3)There were significant differences in the levels of MDA, SOD and GSH in the hippocampus oxidative stress injury indexes among the three groups ( F=87.41, 91.38, 28.69, all P<0.01). The level of MDA in hippocampus of ovariectomized group was higher than that of normal group, and that in the ovariectomized-TR1 group was lower than that of ovariectomized rats group ( P<0.05). And what's more the levels of SOD and GSH in ovariectomized group were lower than those of normal group ( P<0.05), and the ovariectomized-TR1 group was higher than that of ovariectomized group ( P<0.05). (4) The results of Western blot and RT-PCR showed that the levels of p21 and p53 in hippocampal tissue of ovariectomized group were higher than those of normal group ( P<0.05), while the level of aging-related protein p21 and p53 in ovariectomized-TR1 group were significantly lower than those in ovariectomized group ( P<0.05). The level of apoptotic protein Caspase-3 in the hippocampus of ovariectomized rats was higher than that in the normal group ( P<0.05), while the level of Caspase-3 in ovariectomized-TR1 group was significantly lower than that in ovariectomized rats ( P<0.05). The level of anti-apoptotic protein Bcl-2 in hippocampus of ovariectomized group was lower than that of normal rats ( P<0.05), while the level of Bcl-2 in ovariectomized-TR1 group was significantly higher than that in ovariectomized rats ( P<0.05). Conclusion:Overexpression of TR1 can reduce apoptosis of hippocampal cells by regulating oxidative damage and reducing cell senescence.

Persea americana attenuates inflammatory response associated with hyperlipidemia in ovariectomized and irradiated rats by regulating MMP-3/TIMP-1 levels

Objective: To explore the effect of Persea americana supplementation on inflammation, oxidative stress, and lipid profiles in ovariectomized rats fed with a high-fat diet and exposed to radiation. Methods: The control group was sham operated, while groups 2-5 were ovariectomized and fed a high-fat diet. Groups 4 and 5 were exposed to γ-radiation (1 Gy/week for 5 weeks) after ovariectomy. Groups 3 and 5 were treated with 1 mL/250 g/day of Persea americana for one month. Serum levels of estrogen, alanine aminotransferase, aspartate aminotransferase, cholesterol, triglycerides and lipoproteins were measured. Additionally, hepatic oxidative stress, inflammatory and fibrogenic markers were evaluated. Results: Persea americana treatment reduced the oxidative stress markers as well as the levels of triglyceride, total cholesterol, and low-density lipoprotein cholesterol, which in turn lowered hepatic fat accumulation. Moreover, it suppressed hepatic inflammatory mediators (interleukin-6, tumor necrosis factor-α, and C-reactive protein) and downregulated pro-fibrogenic markers (transforming growth factor-β and tissue inhibitor of metalloproteinase-1). Conclusions: Persea americana provides protection against ovariectomy, and gamma radiation-mediated hepatic inflammation not only through its antioxidant, anti-inflammatory, lipid-lowering effect but also by modulating the fibrogenic markers.

Mechanism of Jiawei Bazhen Yimu Capsule in Treatment of Premature Ovarian Failure Based on Estrogen Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the effect of Jiawei Bazhen Yimu capsule on serum sex hormones， sexual organs and estrogen signaling pathway in female rats with premature ovarian failure. MethodThe key target proteins of Jiawei Bazhen Yimu capsule in the treatment of premature ovarian failure were screened out by network pharmacology analysis. Female healthy SD rats were selected, and the rat model of premature ovarian failure was established by ovariectomy. Fifty ovariectomized rats were randomly divided into a model group, an estradiol (E2) valerate group, and Jiawei Bazhen Yimu capsule low, medium, and high-dose groups. Another 10 healthy female rats were set as a sham operation group. The sham operation group and the model group were given distilled water by gavage, and other administration groups were given corresponding doses of drugs by gavage. After 21 d, the serum hormone levels of female rats were measured, including E2, progesterone (P), follicle stimulating hormone (FSH), and luteinizing hormone (LH). Immunofluorescence staining (IF) was used to detect the protein expression levels of estrogen receptor 1 (ESR1), estrogen metabolism P4503A4 enzyme (CYP3A4), and P45019A1 enzyme (CYP19A1) in the uterine tissues of female rats. ResultAs compared with the model group, the serum E2 and P levels of female rats in the Jiawei Bazhen Yimu capsule low, medium, and high-dose groups were significantly increased. Jiawei Bazhen Yimu capsule improved the endometrial status of female rats and increased positive expression of ESR1, CYP3A4, and CYP19A1 in the uterine tissues of female rats (P<0.05). ConclusionThe mechanism of Jiawei Bazhenyimu capsule in the treatment of premature ovarian failure may be related to its hormone-like effect and activation of the estrogen signaling pathway.

Ceiling effect of Postconditioning and Atrial Natriuretic Peptide in Cardioprotection against Ischemia Reperfusion Injury in Ovariectomized rat hearts

Abstract Ischemic postconditioning (IPTC) brings cardioprotection endogenously, Atrial natriuretic peptide (ANP) produces the same effect. It happens due to down expression of endothelial nitric oxide synthase (eNOS). Thus, experimental protocol associating IPTC has been formulated to find the role of ANP in the cardioprotection of heart in OVX rats. For this experiment, heart was isolated from OVX rat and held tightly on Langendorff's apparatus in a manner that ischemia of 30 min and reperfusion of 120 min were also given. Simultaneously, IPTC with four cycles of 5 min ischemia and 5 min reperfusion of each was applied. Parameters like size of myocardial infarct, levels of lactate dehydrogenase (LDH) and release of creatine kinase- MB (CK-MB) in coronary effluent were noted after each stage of experiment for ensuring the extent of myocardial injury. Some significant changes were also seen in the histopathology of cardiovascular tissues. The cardio-protection has been made by four cycles of IPTC. It was confirmed by decline in the size of myocardial infarct. It diminishes the release of LDH and CK-MB in heart of OVX rat. Thus, IPTC induces cardio-protection in the isolated heart from OVX rat. Perfusion of ANP associating with IPTC favors the cardioprotection which is further confirmed by rise in the NO release and heart rate. The level of myocardial damage changes using IPTC, IPTC+OVX, IPTC+OVX+ANP, IPTC+ OVX+ANP+L-NAME and other groups were observed significantly and were found to be less than those in I/R control group. Thus, it is recommended that ANP involving IPTC restores attenuated cardio-protection in OVX rat heart. Therefore, Post-conditioning is useful in various clinical aspects.

Role of ACE and ACE-2 in abrogated cardioprotective effect of ischemic preconditioning in ovariectomized rat heart

Abstract Ischemic heart disease is the leading cause of death in postmenopausal women. The activity of heart ACE increases whereas the activity of ACE-2 decreases after menopause. The present study was designed to investigate the role of ACE and ACE-2 in the abrogated cardioprotective effect of IPC in OVX rat heart. The heart was isolated from OVX rat and mounted on Langendorff's apparatus for giving intermittent cycles of IPC. The infarct size was estimated using TTC stain, and coronary effluent was analyzed for LDH, CK-MB, and nitrite release. IPC induced cardioprotection was significantly attenuated in the ovariectomized rat heart as compared to the normal rat heart. However, this attenuated cardioprotection was significantly restored by perfusion of DIZE, an ACE-2 activator, and captopril, an ACE inhibitor, alone or in combination noted in terms of decrease in myocardial infarct size, the release of LDH and CK-MB, and also increase in the release of NO as compared to untreated OVX rat heart. Thus, it is suggested that DIZE and captopril, alone or in combination restore the attenuated cardioprotective effect of IPC in OVX rat heart which is due to an increase in ACE-2 activity and decrease in ACE activity after treatment.

Alendronate combined with Lactobacillus rhamnosus to combat bone loss in ovariectomized mice / 中华创伤骨科杂志

Objective:To study the protective effect of alendronate combined with Lactobacillus rhamnosus on bone loss in ovariectomized mice.Methods:Fifty female C57BL/6 mice were divided into 5 equal groups ( n=10). Ovariotomy was performed in groups A, B, C and D while a sham operation was performed in group E. Group A was subjected to combined administration of alendronate and Lactobacillus rhamnosus, group B to administration of alendronate, group C to administration of Lactobacillus rhamnosus, and groups D and E to administration of physiological saline only. At 3 months after operation, all the mice were sacrificed to harvest their femurs. Micro CT scanning was performed to detect the bone mineral density (BMD), trabecular relative volume, bone surface area/bone volume, and trabecular thickness and number of trabecular bone. Three-point bending test was used to detect the maximum load, stiffness, ultimate load, Young＇s modulus, and fracture energy. Osteocalcin and alkaline phosphatase levels were measured using blood samples from the mice eyeballs. The 2 groups were compared in terms of all the above indexes. Results:The BMD [(669.87±67.87) mg/cm 3], maximum load [(14.35±0.75) N] and fracture energy [(1,497.43±38.29) J/m 2] in group A were significantly higher than those in group B [(520.07±9.01) mg/cm 3, (11.94±0.82) N and(1,277.61±35.12) J/m 2] and group C [(388.15±25.61) mg/cm 3, (11.10±0.93) N and (1,115.27±63.24) J/m 2] (all P<0.05). The osteocalcin level in group A [(22.25±1.78) ng/mL] was significantly higher than that in group B [(19.08±1.45) ng/mL] and group D [(19.33±1.66) ng/mL] (both P<0.05). The alkaline phosphatase level in group A [(83.21±9.69) ng/mL] was significantly lower than that in group C [(113.16±14.44) ng/mL] and group D [(137.96±14.01) g/mL] (both P<0.05). Conclusion:Alendronate combined with Lactobacillus rhamnosus may play a synergistic role in prevention of bone loss in ovariectomized mice, because combined administration of the two is more effective than administration of either of the two.

Tumores ováricos en pediatría: revisión de los casos en un servicio de pediatría durante 10 años / Pediatric ovarian tumors: a 10-year review in a pediatric service

Los tumores ováricos, a diferencia de lo que sucede en la edad adulta, son infrecuentes en la población pediátrica. Predomina la estirpe germinal, con altas tasas de supervivencia. El objetivo de este estudio es presentar la epidemiología, clínica, diagnóstico y tratamiento de las pacientes de 0-15 años con diagnóstico, entre 2007 y 2017, de tumor ovárico en nuestro centro. Fueron 8 los casos encontrados de 171 tumores diagnosticados (el 4,7 %), con edad media de presentación de 12,5 años. Predominaban, al momento del debut, alteraciones menstruales, dolor abdominal y aumento de perímetro abdominal. Fueron de tipo germinal 6/8, y el teratoma maduro fue el más frecuente. Todas se diagnosticaron con ecografía abdominal, y se confirmó el diagnóstico en 7/8 con resonancia magnética. Se intervinieron todos los casos; predominó la salpingo-ooforectomía, y una paciente precisó quimioterapia adyuvante. La supervivencia libre de enfermedad fue del 100 %.

Unlike adults, ovarian tumors are infrequent in the pediatric population, predominating the germ line at this age, with high survival rates. The objective is to present the epidemiological, clinical, diagnosis and therapeutic characteristics of 0 to 15-year-old patients diagnosed with ovarian tumor in our center between 2007 and 2017.Eight cases out of 171 diagnosed tumors (4.7 %) were found, with a mean age of presentation of 12.5 years. At the moment of diagnosis, menstrual disturbances, abdominal pain and an increase in abdominal circumference predominated. Six out of eight were germ cell tumors, being the mature teratoma the most frequent one. All cases were diagnosed with abdominal ultrasound scan, confirmed in 7/8 cases with magnetic resonance imaging. All cases underwent surgery, predominating salpingo-oophorectomy with one patient requiring adjuvant chemotherapy. Disease-free survival was 100 %.

Análisis descriptivo de un modelo de osteoporosis inducida en ratas (Rattus norvegicus) / Descriptive analysis of an induced osteoporosis model in rats (Rattus norvegicus)

RESUMEN: El objetivo del presente estudio fue realizar un análisis histopatológico de un modelo animal en ratas, que simule las características observables en huesos de pacientes con osteoporosis. Para ello, se utilizaron 10 ratas hembras (Rattus norvegicus) Sprague Dawley de 12 semanas de desarrollo y aproximadamente 200-250 g. De las cuales, a 5 se le realizó ovariectomía (OVX) bilateral, mientras que las 5 restantes fueron utilizadas como control. Posterior a 12 semanas de realizadas las ovariectomías en el grupo experimental, se realizó la eutanasia de los animales y la obtención de ambos fémur, los cuales fueron posteriormente seccionados para procesar su porción distal para su procesamiento histológico de rutina. Todos los animales sobrevivieron al final del estudio sin ninguna complicación postoperatoria, las imágenes histológicas evidenciaron en el grupo experimental (OVX), una disminución del grosor del hueso cortical, mayor cantidad de hueso esponjoso, pérdida de la continuidad de periostio y endostio alrededor de la matriz ósea además de mayor cantidad de tejido adiposo en la médula ósea, al ser comparados con el grupo control. Se puede concluir que a las 12 semanas post ovariectomía se observa un fenotipo histopatológico compatible con características oseteoporóticas en ratas adultas.

SUMMARY: The aim of the present study was to conduct a histopathological analysis of an animal model in rats, which simulates the characteristics observable in bones of patients with osteoporosis. To this end, 10 female rats (Rattus norvegicus) Sprague Dawley of 12 weeks of development and approximately 200-250 g were used. Of these, 5 underwent bilateral ovariectomy (OVX), whereas the remaining 5 were used as control. After 12 weeks of ovariectomy in the experimental group, the animals were euthanized and the two femurs were collected, which were then sectioned to process their distal portion. All the animals survived at the end of the study without any signs of postoperative complications. In the experimental group (OVX), the histological images showed a decrease in the thickness of the cortical bone, a greater amount of cancellous bone, loss of the continuity of the periosteum and endostium around the bone matrix in addition to a greater amount of adipose tissue in the bone marrow, when compared with the control group. It can therefore be inferred that a histopathological phenotype can be found at 12 weeks post- ovariectomy that would be consistent with osteoporotic characteristics in adult rats.

Effect of Lepidium meyenii (maca) on spatial memory and brain oxidative damage of ovariectomised-rats exposed to mobile phone / Efecto de Lepidium meyenii (maca) sobre la memoria espacial y el daño oxidativo cerebral de ratas ovariectomizadas expuestas a teléfonos móviles

Background: Oxidative stress, genetic expression changes, and decomposition of the blood-brain barrier have been shown to be caused by exposure to radiofrequency radiation (RFR); therefore, the search for drugs or food products that offer some protection from these effects has attracted interest. Lepidium meyenii (maca) is a native Andean plant known for its effects on the female reproductive system as well as its immunomodulating, energizing, antioxidant, and nutritive properties due to its high concentration of alkaloids, amino acids, glucosinolates, fatty acids, and macamides. Objectives: We aim to determine the effect of dietary maca supplementation on spatial memory and cerebral oxidative damage in rats with induced menopause and/or with exposure to mobile phone radiation. Method:Female rats divided into 6 groups (six rats in each group): control or not ovariectomized (O-), ovariectomized (O+), ovariectomized + estrogen (O+E+), ovariectomized + estrogen + irradiated (O+E+I+), ovariectomized + maca (O+M+), and ovariectomized + maca + irradiated (O+M+I). Rat spatial memory was tested using the Morris water navigation task. Rats were sacrificed, and cerebral malondialdehyde was measured. Results: After exposure to mobile phone radiation, the group supplemented with maca showed improved spatial memory and lower levels of malondialdehyde than the estrogenized group. Conclusions: Our results suggest that consumption of maca has a positive effect on the maintenance of memory and decreases oxidative stress caused by mobile phone radiation. However, it is necessary to extend this work using a larger sample size

Antecedentes: El estrés oxidativo, los cambios en la expresión genética y la descomposición de la barrera hematoencefálica son causados por la exposición a la radiación de radiofrecuencia (RFR); por tanto, ha despertado interés la búsqueda de fármacos o productos alimenticios que ofrezcan alguna protección frente a estos efectos. Lepidium meyenii (maca) es una planta andina nativa conocida por sus efectos sobre el sistema reproductor femenino así como por sus propiedades inmunomoduladoras, energizantes, antioxidantes y nutritivas debido a su alta concentración de alcaloides, aminoácidos, glucosinolatos, ácidos grasos y macamidas. Objetivos: Determinar el efecto de la suplementación dietética con maca sobre la memoria espacial y el daño oxidativo cerebral en ratas con menopausia inducida y / o con exposición a la radiación de teléfonos móviles. Método: Ratas hembras divididas en 6 grupos (seis ratas en cada grupo): control o no ovariectomizadas (O-), ovariectomizadas (O +), ovariectomizadas + estrógeno (O + E +), ovariectomizadas + estrógeno + irradiadas (O + E + I +), ovariectomizado + maca (O + M +), y ovariectomizado + maca + irradiado (O + M + I). La memoria espacial de las ratas se probó utilizando la tarea de navegación acuática de Morris. Se sacrificaron ratas y se midió el malondialdehído cerebral. Resultados: Después de la exposición a la radiación del teléfono móvil, el grupo suplementado con maca mostró una memoria espacial mejorada y niveles más bajos de malondialdehído que el grupo estrogenizado. Conclusiones: Nuestros resultados sugieren que el consumo de maca tiene un efecto positivo en el mantenimiento de la memoria y disminuye el estrés oxidativo causado por la radiación del teléfono móvil. Sin embargo, es necesario ampliar este trabajo utilizando un tamaño de muestra más grande

Neuroinflammatory effects of tDCS in ovariectomized rats with chronic inflammation

Introduction: Postmenopausal women are more susceptible to chronic conditions, such as osteoporosis, arthritis, and other inflammatory diseases. We investigated the effects of transcranial direct current stimulation (tDCS) on biomarker levels in ovariectomized rats subjected to an inflammatory model. Methods: Twenty adult female Wistar rats underwent ovariectomy and complete Freund's adjuvant (CFA)-induced inflammation. We divided them into 2 groups: OAS (sham tDCS) and OAT (active tDCS). Fifteen days later, the rats underwent bimodal tDCS treatment (20 min, 0.5 mA, 8 days). After 24 h of the last tDCS session, we killed the rats and collected tissue samples (hypothalamus, cerebral cortex, and brainstem) for biomarker analysis by ELISA. We removed the paws for histological analysis. Results: Active tDCS increased hypothalamic and cortical TNF-α and NGF levels, hypothalamic and brainstem IL-1ß levels, and hypothalamic IL-10 levels. Histology of paws showed an inflammatory profile. We observed a small tDCS effect, not statistically significant. Discussion: Bimodal tDCS had an effect on the central inflammatory axis, with a small effect on the peripheral site as evaluated by histology in the current study. (AU)

Do avanafil and zaprinast exert positive effects on bone tissue via the nitric oxide/cyclic guanosine monophosphate/protein kinase-G signaling pathway in rats with ovariectomy-induced osteoporosis?

Phosphodiesterase-5 inhibitors (PDE-5Is) exert positive effects on bone healing and mineralization by activation the nitric oxide/cyclic guanosine monophosphate/protein kinase-G (NO/cGMP/PKG) signaling pathway. In this study, the effects of zaprinast and avanafil, two PDE-5Is, on the NO signaling pathway, estrogen levels, selected bone formation and destruction marker levels, whole-body bone mineral density (WB-BMD), right femur trabecular bone thickness (RF-TBT) and epiphyseal bone width, angiogenesis in the bone-marrow, and selected oxidative stress parameter levels were investigated in rats with ovariectomy-induced osteoporosis. Twenty four adult rats (8 months old) were equally divided into four groups. The first group was the sham operated group. Groups 2, 3 and 4 included ovariectomized rats. At six months after ovariectomy, the 3rd and 4th groups were administered 10 mg/kg zaprinast and avanafil daily as a single dose for 60 days, respectively. Increases in the activity of the NO/cGMP/PKG signalling-pathway, C-terminal collagen peptide levels, angiogenesis in the bone marrow, RF-TBT, epiphyseal bone width and WB-BMD were observed compared to the ovariectomized positive control group (OVX), while the pyridinoline and deoxypyridinoline levels were decreased in the OVX+zaprinast and OVX+avanafil groups (p<0.05). The malondialdehyde, ubiquinone10/ubiquinol10 and 8-hydroxy-2-deoxyguanosine/106deoxyguanosine levels were also increased in the ovariectomized groups compared to the sham group (p<0.05). Based on these results, the levels of bone atrophy and some markers of oxidative stress were increased due to acute estrogen deficiency induced by ovariectomy, but zaprinast and avanafil administration significantly prevented these changes

6-hydroxydopamine and ovariectomy has no effect on heart rate variability parameters of females

OBJECTIVES: In addition to the classic motor symptoms of Parkinson's disease (PD), patients also present with non-motor symptoms, such as autonomic dysfunction, which is present in almost 90% of patients with PD, affecting the quality of life and mortality. Regarding sex differences in prevalence and presentation, there is increasing concern about how sex affects autonomic dysfunction. However, there are no previous data on autonomic cardiac function in females after 6-hydroxydopamine (6-OHDA) striatal injection. METHODS: Wistar female rats were ovariectomized. After 20 days, the animals received bilateral injections of 6-OHDA (total dose per animal: 48 µg) or a vehicle solution in the striatum. Thirty days after 6-OHDA injection, subcutaneous electrodes were implanted for electrocardiogram (ECG) recording. Ten days after electrode implantation, ECG signals were recorded. Analyses of heart rate variability (HRV) parameters were performed, and the 6-OHDA lesion was confirmed by analyzing the number of tyrosine hydroxylase-positive neurons in the substantia nigra pars compacta (SNpc). RESULTS: A high dose of 6-OHDA did not affect HRV of females, independent of ovariectomy. As expected, ovariectomy did not affect HRV or lesions in the SNpc after 6-OHDA injection. CONCLUSIONS: We suggest that females with 6-OHDA present with cardioprotection, independent of ovarian hormones, which could be related to female vagal predominance.