Sex and strain-specific changes of macrophage cytokine profile in female rats during reproductive aging

Starenje se povezuje sa razvojem sistemskog, sterilnog hroničnog zapaljenja (engl. „inflammaging”). Malo je podataka o uticaju genetskih faktora i pola na sposobnost makrofaga (Mφ), kao ključnih ćelija urođenog imunskog odgovora, da tokom starenja “kontrolišu” rezoluciju akutnog zapaljenja i time razvoj hroničnog zapaljenja. Ciljevi ove disertacije su bili da se ispita 1) uticaj rane faze reproduktivnog starenja na fenotipske osobine (ekspresija markera povezanih sa aktivacijom i poreklom/funkcijom) i funkcijska svojstva (fagocitoza, sinteza inflamatornih medijatora) Mφ “mirne” i inflamirane (delovanjem tioglikolata) peritonealne duplje ženki Albino Oxford (AO) pacova, 2) značaj genetskih faktora za reproduktivnim starenjem uslovljene promene peritonealnih Mφ od značaja za uspešnu rezoluciju akutnog zapaljenja i 3) uloga polnih steroida u nastanku ovih promena uporednom analizom promena kod mužjaka i ženki AO pacova, njihovom analizom kod ženki AO pacova kojima su na kraju reproduktivnog perioda uklonjeni jajnici i ispitivanjem delovanja estradiola na Mφ mladih i sredovečnih ženki AO pacova in vitro. Rezultati su pokazali da: 1) se sposobnost Mφ ženki AO pacova da “kontrolišu” rezoluciju akutnog zapaljenja menja već tokom rane faze reproduktivnog starenja, kao i da su ove promene sojno specifične (Mφ sredovečnih ženki AO pacova koje “uspešnije” stare od ženki Dark Agouti pacova pokazuju svojstva koja se mogu povezati sa boljom “kontrolom” inflamacije); 2) su ove promene polno specifične (Mφ sredovečnih ženki imaju svojstva koja ukazuju na veći kapacitet da “kontrolišu” inflamaciju od Mφ mužjaka istog uzrasta) i 3) u nastanku promena relevantnih za sposobnost Mφ ženki AO pacova da “kontrolišu” akutnu inflamaciju važnu ulogu imaju promene u delovanju i estradiola i progesterona. Dodatno, ispitavanja in vitro su ukazala da su za uzrasno zavisne promene u sposobnosti Mφ da “kontrolišu” inflamaciju pored promene u koncentraciji estradiola važne i one u samim Mφ, koje menjaju njihov odgovor na delovanje estradiola.Aging is associated with the development of systemic, sterile chronic inflammation ("inflammaging"). Little is known about the influence of genetic factors and sex on the ability of macrophages (Mφ), as key innate immune cells, to "control" the resolution of acute inflammation and thus the development of chronic inflammation during aging. The objectives of this dissertation were to examine 1) the influence of the early phase of reproductive aging on phenotypic characteristics (expression of markers associated with activation and origin/function) and functional characteristics (phagocytosis, synthesis of inflammatory mediators) of Mφ isolated from “naive” and inflamed (thioglycollate-induced) peritoneal cavity of females Albino Oxford (AO) rats; 2) the significance of genetic factors for reproductive aging-related changes of peritoneal Mφ, especially those important for successful resolution of acute inflammation and 3) the role of sex steroids in the occurrence of these changes by comparative analysis in males and females of AO rats, their analysis in female AO rats whose ovaries were removed at the end of the reproductive period and by examining in vitro effect of estradiol on Mφ from young and middle-aged female AO rats. The results showed that: 1) the ability of Mφ from AO females to “control” the resolution of acute inflammation changes during the early phase of reproductive aging, and these changes are strain-specific (Mφ from middle-aged AO females that “age more successful” than Dark Agouti females, show properties which may be associated with better "control" of inflammation); 2) these changes are sex-specific (Mφ from middle-aged females have a greater capacity to “control” inflammation than Mφ from males of the same age group) and 3) both estradiol and progesterone play important roles in the ability of Mφ from AO females to “control” acute inflammation. In vitro studies revealed that, in addition to changes in estradiol concentration, intrinsic changes in Mφ that regulate their response to estradiol action are important for age- dependent changes in Mφ ability to „control“ inflammation

Sex and strain-specific changes of macrophage cytokine profile in female rats during reproductive aging

Starenje se povezuje sa razvojem sistemskog, sterilnog hroničnog zapaljenja (engl.„inflammaging”). Malo je podataka o uticaju genetskih faktora i pola na sposobnost makrofaga(Mφ), kao ključnih ćelija urođenog imunskog odgovora, da tokom starenja “kontrolišu”rezoluciju akutnog zapaljenja i time razvoj hroničnog zapaljenja. Ciljevi ove disertacije su bilida se ispita 1) uticaj rane faze reproduktivnog starenja na fenotipske osobine (ekspresijamarkera povezanih sa aktivacijom i poreklom/funkcijom) i funkcijska svojstva (fagocitoza,sinteza inflamatornih medijatora) Mφ “mirne” i inflamirane (delovanjem tioglikolata)peritonealne duplje ženki Albino Oxford (AO) pacova, 2) značaj genetskih faktora zareproduktivnim starenjem uslovljene promene peritonealnih Mφ od značaja za uspešnurezoluciju akutnog zapaljenja i 3) uloga polnih steroida u nastanku ovih promena uporednomanalizom promena kod mužjaka i ženki AO pacova, njihovom analizom kod ženki AO pacovakojima su na kraju reproduktivnog perioda uklonjeni jajnici i ispitivanjem delovanjaestradiola na Mφ mladih i sredovečnih ženki AO pacova in vitro. Rezultati su pokazali da: 1) sesposobnost Mφ ženki AO pacova da “kontrolišu” rezoluciju akutnog zapaljenja menja većtokom rane faze reproduktivnog starenja, kao i da su ove promene sojno specifične (Mφsredovečnih ženki AO pacova koje “uspešnije” stare od ženki Dark Agouti pacova pokazujusvojstva koja se mogu povezati sa boljom “kontrolom” inflamacije); 2) su ove promene polnospecifične (Mφ sredovečnih ženki imaju svojstva koja ukazuju na veći kapacitet da“kontrolišu” inflamaciju od Mφ mužjaka istog uzrasta) i 3) u nastanku promena relevantnih zasposobnost Mφ ženki AO pacova da “kontrolišu” akutnu inflamaciju važnu ulogu imajupromene u delovanju i estradiola i progesterona. Dodatno, ispitavanja in vitro su ukazala da suza uzrasno zavisne promene u sposobnosti Mφ da “kontrolišu” inflamaciju pored promene ukoncentraciji estradiola važne i one u samim Mφ, koje menjaju njihov odgovor na delovanjeestradiola.Aging is associated with the development of systemic, sterile chronic inflammation("inflammaging"). Little is known about the influence of genetic factors and sex on the abilityof macrophages (Mφ), as key innate immune cells, to "control" the resolution of acuteinflammation and thus the development of chronic inflammation during aging. The objectivesof this dissertation were to examine 1) the influence of the early phase of reproductive agingon phenotypic characteristics (expression of markers associated with activation andorigin/function) and functional characteristics (phagocytosis, synthesis of inflammatorymediators) of Mφ isolated from “naive” and inflamed (thioglycollate-induced) peritonealcavity of females Albino Oxford (AO) rats; 2) the significance of genetic factors forreproductive aging-related changes of peritoneal Mφ, especially those important forsuccessful resolution of acute inflammation and 3) the role of sex steroids in the occurrence ofthese changes by comparative analysis in males and females of AO rats, their analysis infemale AO rats whose ovaries were removed at the end of the reproductive period and byexamining in vitro effect of estradiol on Mφ from young and middle-aged female AO rats. Theresults showed that: 1) the ability of Mφ from AO females to “control” the resolution of acuteinflammation changes during the early phase of reproductive aging, and these changes arestrain-specific (Mφ from middle-aged AO females that “age more successful” than Dark Agoutifemales, show properties which may be associated with better "control" of inflammation); 2)these changes are sex-specific (Mφ from middle-aged females have a greater capacity to“control” inflammation than Mφ from males of the same age group) and 3) both estradiol andprogesterone play important roles in the ability of Mφ from AO females to “control” acuteinflammation. In vitro studies revealed that, in addition to changes in estradiol concentration,intrinsic changes in Mφ that regulate their response to estradiol action are important for age-dependent changes in Mφ ability to „control“ inflammation

Sex and strain-specific changes of macrophage cytokine profile in female rats during reproductive aging

Starenje se povezuje sa razvojem sistemskog, sterilnog hroničnog zapaljenja (engl. „inflammaging”). Malo je podataka o uticaju genetskih faktora i pola na sposobnost makrofaga (Mϕ), kao ključnih ćelija urođenog imunskog odgovora, da tokom starenja “kontrolišu” rezoluciju akutnog zapaljenja i time razvoj hroničnog zapaljenja. Ciljevi ove disertacije su bili da se ispita 1) uticaj rane faze reproduktivnog starenja na fenotipske osobine (ekspresija markera povezanih sa aktivacijom i poreklom/funkcijom) i funkcijska svojstva (fagocitoza, sinteza inflamatornih medijatora) Mϕ “mirne” i inflamirane (delovanjem tioglikolata) peritonealne duplje ženki Albino Oxford (AO) pacova, 2) značaj genetskih faktora za reproduktivnim starenjem uslovljene promene peritonealnih Mϕ od značaja za uspešnu rezoluciju akutnog zapaljenja i 3) uloga polnih steroida u nastanku ovih promena uporednom analizom promena kod mužjaka i ženki AO pacova, njihovom analizom kod ženki AO pacova kojima su na kraju reproduktivnog perioda uklonjeni jajnici i ispitivanjem delovanja estradiola na Mϕ mladih i sredovečnih ženki AO pacova in vitro. Rezultati su pokazali da: 1) se sposobnost Mϕ ženki AO pacova da “kontrolišu” rezoluciju akutnog zapaljenja menja već tokom rane faze reproduktivnog starenja, kao i da su ove promene sojno specifične (Mϕ sredovečnih ženki AO pacova koje “uspešnije” stare od ženki Dark Agouti pacova pokazuju svojstva koja se mogu povezati sa boljom “kontrolom” inflamacije); 2) su ove promene polno specifične (Mϕ sredovečnih ženki imaju svojstva koja ukazuju na veći kapacitet da “kontrolišu” inflamaciju od Mϕ mužjaka istog uzrasta) i 3) u nastanku promena relevantnih za sposobnost Mϕ ženki AO pacova da “kontrolišu” akutnu inflamaciju važnu ulogu imaju promene u delovanju i estradiola i progesterona. Dodatno, ispitavanja in vitro su ukazala da su za uzrasno zavisne promene u sposobnosti Mϕ da “kontrolišu” inflamaciju pored promene u koncentraciji estradiola važne i one u samim Mϕ, koje menjaju njihov odgovor na delovanje estradiola.Aging is associated with the development of systemic, sterile chronic inflammation ("inflammaging"). Little is known about the influence of genetic factors and sex on the ability of macrophages (Mϕ), as key innate immune cells, to "control" the resolution of acute inflammation and thus the development of chronic inflammation during aging. The objectives of this dissertation were to examine 1) the influence of the early phase of reproductive aging on phenotypic characteristics (expression of markers associated with activation and origin/function) and functional characteristics (phagocytosis, synthesis of inflammatory mediators) of Mϕ isolated from “naive” and inflamed (thioglycollate-induced) peritoneal cavity of females Albino Oxford (AO) rats; 2) the significance of genetic factors for reproductive aging-related changes of peritoneal Mϕ, especially those important for successful resolution of acute inflammation and 3) the role of sex steroids in the occurrence of these changes by comparative analysis in males and females of AO rats, their analysis in female AO rats whose ovaries were removed at the end of the reproductive period and by examining in vitro effect of estradiol on Mϕ from young and middle-aged female AO rats. The results showed that: 1) the ability of Mϕ from AO females to “control” the resolution of acute inflammation changes during the early phase of reproductive aging, and these changes are strain-specific (Mϕ from middle-aged AO females that “age more successful” than Dark Agouti females, show properties which may be associated with better "control" of inflammation); 2) these changes are sex-specific (Mϕ from middle-aged females have a greater capacity to “control” inflammation than Mϕ from males of the same age group) and 3) both estradiol and progesterone play important roles in the ability of Mϕ from AO females to “control” acute inflammation. In vitro studies revealed that, in addition to changes in estradiol concentration, intrinsic changes in Mϕ that regulate their response to estradiol action are important for agedependent changes in Mϕ ability to „control“ inflammation

Lactobacillus rhamnosus Affects Rat Peritoneal Cavity Cell Response to Stimulation with Gut Microbiota: Focus on the Host Innate Immunity

Gut microbiota contribute to shaping the immune repertoire of the host, whereas probiotics may exert beneficial effects by modulating immune responses. Having in mind the differences in both the composition of gut microbiota and the immune response between rats of Albino Oxford (AO) and Dark Agouti (DA) rat strains, we investigated if intraperitoneal (i.p.) injection of live Lactobacillus rhamnosus (LB) may influence peri-toneal cavity cell response to invitro treatments with selected microbiota in the rat strain-dependent manner. Peritoneal cavity cells from AO and DA rats were lavaged two (d2) and seven days (d7) following i.p. injection with LB and tested for NO, urea, and H2O2 release basally, or upon invitro stimulation with autologous E.coli and Enterococcus spp. Whereas the single i.p. injection of LB nearly depleted resident macrophages and increased the proportion of small inflammatory macrophages and monocytes on d2 in both rat strains, greater proportion of MHCIIhiCD163− and CCR7+ cells and increased NO/diminished H2O2 release in DA compared with AO rats suggest a more intense inflammatory prim-ing by LB in this rat strain. Even though E.coli- and/or Enterococcus spp.-induced rise in H2O2 release invitro was abrogated by LB in cells from both rat strains, LB prevented microbiota-induced increase in NO/urea ratio only in cells from AO and augmented it in cells from DA rats. Thus, the immunomodulatory properties may not be constant for particular probiotic bacteria, but shaped by innate immunity of the host.https://rdcu.be/cxyn

17β-Estradiol and genistein affect macrophage inflammatory cytokine production during aging in sex-specific manner

Aging differently affects the expression of estrogen receptors alpha and beta (ERα and ERβ) and Toll-like receptors (TLR4) on peritoneal cavity cells of male and female rats. We explored the involvement of ERα and ERβ in the in vitro treatment of LPS-stimulated peritoneal macrophages with 17β-estradiol (which stimulates both receptors) or genistein (which is predominantly an ERβ agonist) on inflammatory cytokine secretion from young (3 months old) and middle-aged (16 months old) female and male AO rats. Aging diminished the proportion of TLR4+ cells and secretion of IL-1β and IL-6 in macrophages from female rats while the effect on male rat macrophages was opposite. 17βestradiol increased IL-1β secretion by middle-aged females’ macrophages via ERα, and suppressed it in cells from young females via ERβ. Genistein-induced decrease of IL-1β in macrophages from all experimental groups was probably mediated by ERβ. 17β-estradiol augmented IL-6 secretion by cells from all experimental groups via ERα while genistein diminished it in all females’ and in middle-aged male rats’ macrophages by activating ERβ. However, genistein increased IL-6 secretion from macrophages of young male rats via ERα. Although 17β-estradiol and genistein stimulated secretion of macrophage inflammatory cytokines via ERα and suppressed it probably via ERβ, their modulatory actions were determined by aging-induced changes in macrophage ERs expression and possible ERα / ERβ interactions (Supported by Ministry of Education, Science and Technological development, Republic of Serbia, Grant No 175050)

Potential impact of early-life probiotic supplementation on peritoneal macrophage function

Clinical and animal trials show that early life probiotic consumption provides health benefits in adult life by modulating the immune response. We tested the effects of early life oral consumption of the probiotic Lactobacillus rhamnosus on the function and phenotype of rat peritoneal cavity cells in a model of induced colitis. For the first month of their lives, rats were either fed with an aqueous probiotic bacteria suspension (LB group) or tap water (control group). When the rats grew to 3 months old, we studied the response of their peritoneal macrophages to autologous fecal bacteria stimulation in vitro, both before and after colitis induction (TNBS 40mg/kg of body mass in 50% ethanol). Compared to the controls, the peritoneal cavity cells of the LB group produced less nitric oxide (NO) and had an increased proportion of CD163+ cells. The rats in the LB group have shown milder symptoms of colitis (shorter length of colon under necrosis, less severe submucosal infiltration, lesser degree of colonic wall thickening), along with a diminished increase of peritoneal proinflammatory CCR7+ cells and blunted NO production in response to stimulation by autologous fecal bacteria. Our results may indicate that early oral probiotic administration attenuates macrophage responses to fecal bacteria, which are the primary cause of tissue inflammation and necrosis in chemically induced colitis models, and that this attenuation may be involved in improving the health of colitic rats

Aging affects rat inflammatory peritoneal exudate composition and macrophage inflammatory mediator production in a strain-dependent manner

The present study was designed to examine influence of aging on macrophage proinflammatory/anti-inflammatory capacity in rat model of thioglycollate-induced peritonitis. Peritoneal macrophages were isolated from young (3-months-old) and aged (18-months-old) Dark Agouti (DA) and Albino Oxford (AO) rats seven days post-injection of thioglycollate medium. Freshly isolated peritoneal exudate cells were examined for the expression of CD163, CCR7, CD14 and TLR4, whereas cytokine production (TNF-α, IL-6 and IL-10) and arginine metabolism end-products (NO and urea) were assayed in vitro under basal conditions and following stimulation with LPS. In DA rat inflammatory peritoneal exudate, aging diminished the frequency of cells with a “resolving macrophage” CD14+CD163+ phenotype. However, in AO rats, which exhibited stable frequency of CD14+CD163+ cells in inflammatory peritoneal exudate with aging, the proportion of CCR7-bearing peritoneal cells, presumably immigrating inflammatory monocytes, was diminished in aged animals. Under basal culture conditions, macrophages from aged rats of both strains released less amount of TNF-α, IL-6 and IL-10, but produced more urea than cells from young strain-matched rats. However, these changes were more pronounced in peritoneal macrophages from AO rats. Additionally, age-related decrease in the frequency of TLR4-expressing cells was observed among fresh peritoneal exudate cells from AO rats. Upon LPS stimulation, the production of prototypic inflammatory cytokines (TNF-α and IL-6) was diminished in macrophages from aged AO rats, whereas aging had the opposite effect on their production in DA rat macrophages. Moreover, aging increased NO production in LPS-stimulated macrophages from DA rats, whereas urea production was enhanced in macrophages from both strains, but this increase was strikingly more pronounced in macrophages from AO rats. Collectively, results suggest that aging affects inflammatory peritoneal exudate cellular composition and macrophage proinflamatory/immunomodulatory capacity in a strain- specific manne

NMDA Receptor Antagonist Memantine Ameliorates Experimental Autoimmune Encephalomyelitis in Aged Rats

Aging is closely related to the main aspects of multiple sclerosis (MS). The average age of the MS population is increasing and the number of elderly MS patients is expected to increase. In addition to neurons, N-methyl-D-aspartate receptors (NMDARs) are also expressed on non-neuronal cells, such as immune cells. The aim of this study was to investigate the role of NMDARs in experimental autoimmune encephalomyelitis (EAE) in young and aged rats. Memantine, a non-competitive NMDAR antagonist, was administered to young and aged Dark Agouti rats from day 7 after immunization. Antagonizing NMDARs had a more favourable effect on clinical disease, reactivation, and apoptosis of CD4+ T cells in the target organ of aged EAE rats. The expression of the fractalkine receptor CX3CR1 was increased in memantine-treated rats, but to a greater extent in aged rats. Additionally, memantine increased Nrf2 and Nrf2-regulated enzymes’ mRNA expression in brain tissue. The concentrations of superoxide anion radicals, malondialdehyde, and advanced oxidation protein products in brain tissue were consistent with previous results. Overall, our results suggest that NMDARs play a more important role in the pathogenesis of EAE in aged than in young rats

Age-Dependent Role Of Nmda Receptors In Experimental Autoimmune Encephalomyelitis

Aims Ageing affects N-methyl-D-aspartate receptors (NMDARs), their expression and function in neuronal and non-neuronal cells. Contribution of NMDARs to pathogenesis of experimental autoimmune encephalomyelitis (EAE) has been investigated but further study is still needed. The aim of this study was to determine whether ageing affects the role of NMDARs in EAE. Methods Memantine, a non-competitive NMDAR antagonist which limits pathological activity of NMDARs while sparing normal synaptic activity, was administered orally from day 7 after immunization to 3- and 24-month-old female Dark Agouti rats. The animals were sacrificed at the peak of the disease. Spinal cord mononuclear cells were analyzed by flow cytometry. Brain tissue was collected for biochemical analysis of redox status and RT-qPCR. Results Semiprophylactic administration of memantine ameliorated clinical disease course, with greater effect in aged rats. Memantine reduced the number, frequency, and reactivation of CD4+ T lymphocytes and increased the relative percentage of CX3CR1-expressing microglia in spinal cord, but to a greater extent in aged rats. Additionally, analysis of brain redox status parameters showed that memantine was more effective in reducing superoxide anion radical, malondialdehyde and advanced oxidation protein products in aged rats than in young ones. In accordance with previous findings, NMDAR inhibition by memantine decreased NADPH oxidase and IL-1β expression and increased the nuclear factor erythroid 2-related factor 2 and heme oxygenase-1 expression, to a greater extent in aged rats. Conclusions The involvement of NMDARs in the pathogenesis of EAE was age-dependent, being more pronounced in aged than in young rats

Age-dependent role of NMDA receptors in experimental autoimmune encephalomyelitis

Aims Ageing affects N-methyl-D-aspartate receptors (NMDARs), their expression and function in neuronal and non-neuronal cells. Contribution of NMDARs to pathogenesis of experimental autoimmune encephalomyelitis (EAE) has been investigated but further study is still needed. The aim of this study was to determine whether ageing affects the role of NMDARs in EAE. Methods Memantine, a non-competitive NMDAR antagonist which limits pathological activity of NMDARs while sparing normal synaptic activity, was administered orally from day 7 after immunization to 3- and 24-month-old female Dark Agouti rats. The animals were sacrificed at the peak of the disease. Spinal cord mononuclear cells were analyzed by flow cytometry. Brain tissue was collected for biochemical analysis of redox status and RT-qPCR. Results Semiprophylactic administration of memantine ameliorated clinical disease course, with greater effect in aged rats. Memantine reduced the number, frequency, and reactivation of CD4+ T lymphocytes and increased the relative percentage of CX3CR1-expressing microglia in spinal cord, but to a greater extent in aged rats. Additionally, analysis of brain redox status parameters showed that memantine was more effective in reducing superoxide anion radical, malondialdehyde and advanced oxidation protein products in aged rats than in young ones. In accordance with previous findings, NMDAR inhibition by memantine decreased NADPH oxidase and IL-1β expression and increased the nuclear factor erythroid 2-related factor 2 and heme oxygenase-1 expression, to a greater extent in aged rats. Conclusions The involvement of NMDARs in the pathogenesis of EAE was age-dependent, being more pronounced in aged than in young rats.FENS Forum/2022, E- Book of Abstract