Aging Model Group Research Articles

Galactose-induced Aging Model in Rat Testicular Tissue.

To examine whether the D-galactose induced aging model is an appropriate model for further aging research. Experimental study. Aziz Sancar Institute of Experimental Medicine, Istanbul University, Turkey, June 2015- June 2017. The study comprises 3 groups of rats. Group I is young control (YC) 5-month-old rats. Group II is 5-month- old rats, which were mimetically aged (MA) for 6 weeks via intraperitoneal D-galactose (60 mg/kg body weight/day, 0.5 mL) administration. Group III is naturally aged (NA) 24-month-old rats. Group I and III received intraperitoneal saline (0.9% 0.5 mL) for 6 weeks as vehicle. Group I and Group II received injections at 21 weeks age and Group III rats 6 weeks before 24 months age. Tissues were harvested when rats became 6.5-month-old (Group I and Group II) and 24-month-old (Group III). Quantitative biochemical analyses of proteins, lipids, DNA biomarkers and Cu, Zn-SOD were conducted. Statistical analysis of the data was conducted using ANOVA, followed by post-hoc Bonferroni test. Higher magnitude of oxidative damage and diminished antioxidant defence capacity were found in both mimetically aged and naturally aged testicular tissues. It is observed that D-galactose aging model group shares significant similarities in terms of impaired redox homeostasis with the naturally aged rats. D-galactose induced testicular aging model successfully mimics aging process. Therefore, D-galactose induced aging model may be used as an accelerated aging model to study the age related alterations and interventions.

Electroacupuncture at Guanyuan (CV 4), Zusanli (ST 36) and Baihui (DU 20) regulate the aging-related changes in gene expression profile of the hippocampus in sub-acutely aging rats.

To investigate the molecular mechanisms of sub-acutely aging and demonstrate the effect of electroacupuncture (EA) at the Guanyuan (CV 4), Zusanli (ST 36) and Baihui (DU 20) acupoint on the sub-acutely aging brain, cDNA microarrays and bioinformatics analyses were carried out. Thirty Sprague-Dawley (SD) male rats were selected and randomly divided into three groups: the control group (C), the sub-acutely aging model group (M) and the electroacupuncture group (M+EA). Sub-acutely aging model rats were obtained by D-galactose s.c. injection continuously for 40 days. Total RNA was extracted from the hippocampus area of brains in three groups for cDNA microarrays. The data of different groups were compared and analyzed by differential expression analysis, Gene ontology (GO) term enrichment, Kyoto Encyclopedia of Genes Genomes (KEGG) pathway enrichment and quantitative real-time PCR. According to the results, 4052 DE genes were identified in our study. Among them, there were 3079 differentially expressed (DE) genes between group M and group C, and these genes are associated with the aging of rats. Moreover, 983 genes were expressed differently in group M+EA compared with group M, revealing that points stimuli could regulate gene expression in brain with aging. Gene ontology (GO) term enrichment and KEGG enrichment were performed to further classify the differential expression genes. Important GO terms and KEGG pathways connected with sub-acutely aging EA effects were identified. At last, 3 significant differentially expressed genes were selected for real-time quantitative PCR to clarify the cDNA microarray results. In conclusion, the cDNA microarray data first compared and analyzed the differences of gene expression profile in the hippocampus of rats in different groups, which contribute to our knowledge on the molecular mechanisms of EA towards sub-acutely aging.

The Effects of Two Different Multivitamins on Aging Mice.

Investigating anti-aging factors that is more effective than antioxidants has important theoretical significance and application value. In search for nutritional ingredients that are more effective in anti-aging, two different multivitamins (multivitamin-1 at a dose of 2.5 mg/kg body weight [BW]/day; multivitamin-2 at a dose of 5.4 mg/kg BW/day) were administered to aging mice (N = 40) induced by D-galactose. The content or activity of the biochemical components associated with aging and anti-aging in the brain and the liver of the experimental mice was then determined for analysis of statistically significant difference among the groups. Results showed the mice in the aging model group exhibited obvious senility symptoms. However, the mice in the multivitamin-1 and multivitamin-2 groups were essentially similar to those of the control group, but were obviously better than the mice in the aging model group. Multivitamin-1 and multivitamin-2 decreased significantly the malondialdehyde (MDA) content and monoamine oxidase (MAO) activity (P < 0.01), and increased significantly the activity of glutathione peroxidase (GSH-Px), copper/zinc-superoxide dismutase (Cu/Zn-SOD) and manganese-superoxide dismutase (Mn-SOD) (P < 0.01) in the brain and the liver of the aging mice. There was no significant difference (P > 0.05) between the effects of the two multivitamins on the components associated with aging and anti-aging. In conclusions, this work showed that vitamins B₁, B₂, B₆ and PP (Nicotinic acid or vitamin B₃) play key roles in the anti-aging process of multivitamin-2. Vitamins B₁, B₂, B₆, and PP are more effective nutrients in anti-aging in mice.

Total Flavonoid Extract from Abelmoschus manihot (L.) Medic Flowers Attenuates d-Galactose-Induced Oxidative Stress in Mouse Liver Through the Nrf2 Pathway.

Abelmoschus manihot (L.) Medic is an edible hibiscus that is rich in flavonoids, and its use as Chinese herbal medicine for the treatment of diseases and health maintenance dates back to ancient times. The chemical compositions of total flavonoid of A. manihot (L.) Medic flower extract (TFAE) were identified and determined by high performance liquid chromatography (HPLC). The effects of TFAE on antioxidative activities in a d-galactose (d-gal)-induced mouse model and Nrf2-mediated antioxidant responses were evaluated. Male Kunming mice were randomly divided into normal control group, d-gal aging model group, d-gal+ascorbic acid group that served as a positive control, and d-gal+TFAE (40, 80, and 160 mg TFAE/kg) group. After 42 days, the antioxidant effects of these treatments were determined by biochemical studies, Western blotting, quantitative real-time polymerase chain reaction, and histological analysis. The results showed that the groups administered TFAE exhibited significant elevation in liver activities of antioxidant enzymes, including catalase (CAT), glutathione peroxidase (GPx), superoxide dismutase (SOD), and total antioxidant capacity (T-AOC), and decreased malondialdehyde (MDA) production in a dose-dependent manner compared with the d-gal-induced model group. Expression of Nrf2 and its target antioxidants (HO-1 and NQO1) was manifestly increased by TFAE treatment. TFAE also increased mRNA expression of GPx, SOD, and CAT and decreased tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β). Furthermore, the microstructure of livers in TFAE-administered mice was obviously improved as compared with the d-gal model group. These results suggest that TFAE protects mice against d-gal-induced oxidative stress, and the effect is related to the activation of Nrf2 signaling.

Potential of bone marrow mesenchymal stem cells in rejuvenation of the aged skin of rats.

The aim of the present study was to evaluate the anti-aging effects of bone marrow-mesenchymal stem cells (BM-MSCs) in a D-galactose-induced skin aging rat model. Male Sprague Dawley rats were randomly divided into four groups (n=10/group) as follows: Normal control group; skin aging model group; MSC-treated group by subcutaneous multi-point injection. The skin aging model was established by a daily subcutaneous injection of 15% D-galactose (1,000 mg/kg) for 8 weeks. Rats in the MSC-treated groups were administered 3×106/ml BM-MSCs/green fluorescent protein (GFP) for 4 weeks, administered once per week. Oxidative/antioxidative parameters were evaluated, and morphological and ultrastructure analyses were performed. Rats in the model group exhibited the typical changes of aging skin. Compared with the control group, rats in the model group had significantly increased malondialdehyde (MDA) content (P<0.01), and decreased serum superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities (P<0.05). MSC treatment markedly ameliorated aging-induced oxidative stress in the skin. Histologically, rats in the model group exhibited loosely arranged epidermal cell layers and disorganized collagen fibers. BM-MSC treatment significantly improved the histological abnormalities, which was similar to those in the control group. In addition, 7 days after the final cell transplantation, GFP-positive cells were observed by fluorescence microscopy to be distributed in the dermis. Injection of BM-MSCs significantly improved the D-galactose-induced histological abnormalities of the skin, by promoting an antioxidant response and ameliorating oxidative stress in aged skin. Thus, BM-MSCs may be beneficial in the rejuvenation of aged skin.

The antioxidant effect of Asparagus cochinchinensis (Lour.) Merr. shoot in d-galactose induced mice aging model and in vitro

BackgroundAn increasing number of plant components and their extracts have been shown to have beneficial health effects in humans. We aimed to explore the antioxidant effects of the aqueous extract of Asparagus cochinchinensis (Lour.) Merr. shoot in vivo and in vitro. MethodsA total of 80 Kun Ming mice were randomly divided into four groups (20/group). The mice in the control group received a daily subcutaneous injection of saline. A daily injection of D-galactose was administered to the aging model group, the vitamin C (Vc) group (positive control group), and the extract treatment group. Regular measurement of blood cells, nitric oxide synthase (NOS), catalase (CAT) activities, superoxide dismutase (SOD) activities, nitric oxide (NO), and malondialdehyde (MDA) concentration, and the expressions of NOS, SOD, and glutathione peroxidase (GPX) in serum levels were obtained. Furthermore, the microstructure of mice viscera was observed using hematoxylin and eosin staining. ResultsThe aqueous extract of A. cochinchinensis (Lour.) Merr. had similar 1,1-diphenyl-2-picrylhydrazyl radical 2,2-diphenyl-1-(2,4,6-trinitrophenyl) hydrazyl (DPPH·) [or 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS+)] and higher hydroxyl radicals (or superoxide anion; p < 0.05) radical scavenging capabilities to Vc. Moreover, compared with the aging model group, the aqueous extract of A. cochinchinensis (Lour.) Merr. shoot could obviously increase NOS, CAT, and SOD activities and the NO content, and reduce the MDA content (p < 0.05). Additionally, the microstructure of mice viscera was obviously improved and the expressions of NOS, SOD and GPX were also manifestly increased in the treatment group (p < 0.05). ConclusionThe aqueous extract of A. cochinchinensis (Lour.) Merr. shoot had a strong radical scavenging capability in vivo and in vitro, and might be used to diminish radicals in the body and consequently prevent aging.

Research on the Influence of Anti-fatigue Effect and Movement Ability of Blueberry Polysaccharides on Aged Mice.

This topic introduces functions, extraction and purification of polysaccharides and focuses on the effect of blue polysaccharides (BPs) on sport capacity, anti-fatigue effect and related physiological indexes in aged mice induced by D-galactose was investigated. Water extract-alcohol precipitation method was used in the experiment to extract polysaccharides from blueberry, after preliminary purification, research on polysaccharide of its vitro antioxidant activity and bacteriostasis is taken, to provide a reference for the production and application of blueberry polysaccharides. Mice were randomly divided into normal control group, aged model group, low-dose BP group, middle-dose BP group, and high-dose BP group. After the administration of BPs for 20 consecutive days, the exhaustive swimming time and fatigue indexes were determined. The results showed that BPs could extend the exhaustive swimming time, decrease the content of BUN, BLA and MDA, and obviously increase the reservation of HG, MG, SOD and LDH. To conclude, BPs has good anti-fatigue effect. This study may provide theoretical evidence for the development of anti-fatigue drugs from BPs.

脑舒胶囊对衰老小鼠免疫功能的调节作用

Objective: To observe the effects of NaoShu capsule on immune function of aging mice . Methods: Aging mice was induced by subcutaneous injections model was induced by subcutaneous injections with 5% D-galactose at the neck for 6 weeks, IL-2 and IL-6 content in serum were detected by means of radio-immunity, and calculating the index of organ by a ratio of the weight of thymus and spleen and the weight of mice. Results: Compared with the control group, the thymus and spleen index of mice was decreased obvious; the IL-2 content was obvious decreased, and the IL-6 content was obvious increased in aging model group. Compared with the model group, the the IL-2 content was enhanced and the IL-6 content was obvious decreased in the therapeutic group with 1.33g/kg and 2.66g/kg Naoshu capsule ( P <0.05, 0.01). Conclusion: There has a certain correlation between aging and immune, and there are obvious accommodation of Naoshu capsule on immune function of aging mice.

Research of anti-aging mechanism of ginsenoside Rg1 on brain

Neurodegenerative disease is common and frequently occurs in elderly patients. Previous studies have shown that ginsenoside Rg1 was able to inhibit senescent of brain, but the mechanism on the brain during the treatment remains elucidated. To study the mechanism of ginsenoside Rg1 in the process of anti-aging of brain, forty male SD rats were randomly divided into normal group, Rg1 normal group, brain aging model group and Rg1 brain aging model group, each group with 10 rats (brain aging model group: subcutaneous injection of D-galactose (120 mg kg(-1)), qd for 42 consecutive days; Rg1 brain aging model group: while copying the same test as that of brain aging model group, begin intraperitoneal injection of ginsenosides Rg1 (20 mg x kg(-1)) qd for 27 d from 16 d. Rg1 normal group: subcutaneous injection of the same amount of saline; begin intraperitoneal injection of ginsenosides Rg1 (20 mg x kg(-1)) qd for 27 d from 16 d. Normal: injected with an equal volume of saline within the same time. Perform the related experiment on the second day after finishing copying the model or the completion of the first two days of drug injections). Learning and memory abilities were measured by Morris water maze. The number of senescent cells was detected by SA-beta-Gal staining while the level of IL-1 and IL-6 proinflammatory cytokines in hippocampus were detected by ELISA. The activities of SOD, contents of GSH in hippo- campus were quantified by chromatometry. The change of telomerase activities and telomerase length were performed by TRAP-PCR and southern blotting assay, respectively. It is pointed that, in brain aging model group, the spatial learning and memory capacities were weaken, SA-beta-Gal positive granules increased in section of brain tissue, the activity of antioxidant enzyme SOD and the contents of GSH decreased in hippocampus, the level of IL-1 and IL-6 increased in hippocampus, while the length of telomere and the activity of telomerase decreased in hippocampus. Rats of Rg1 brain aging group had their spatial learning and memory capacities enhanced, SA-beta-Gal positive granules in section of brain tissue decreased, the activity of antioxidant enzyme SOD and the contents of GSH increased in hippocampus, the level of IL-1 and IL-6 in hippocampus decreased, the length contraction of telomere suppressed while the change of telomerase activity increased in hippocampus. Compared with that of normal group, the spatial learning and memory capacities were enhanced in Rg1 normal group, SA-beta-Gal positive granules in section of brain tissue decreased in Rg1 normal group, the level of IL-1 and IL-6 in hippocampus decreased in Rg1 normal group. The results indicated that improvement of antioxidant ability, regulating the level of proinflammatory cytokines and regulation of telomerase system may be the underlying anti-aging mechanism of Ginsenoside Rg1.

Lactobacillus plantarum NDC 75017 alleviates the learning and memory ability in aging rats by reducing mitochondrial dysfunction.

The aim of the present study was to investigate the protective effect of Lactobacillus plantarum NDC 75017 on D-galactose (D-gal)-induced mitochondrial dysfunction in the rat cerebral cortex. Fifty rats were randomly divided into five groups (n=10 in each group). The rats in the aging model group were subcutaneously injected with 100 mg/kg D-gal and those in the protective groups were additionally orally administered L. plantarum NDC 75017 at doses of 1×108, 1×109 or 1×1010 CFU/100 mg body weight/day, respectively. The control rats were administrated an equal volume of the vehicle. Following continuous treatment for seven weeks, the learning and memory abilities and mitochondrial ultrastructure, function and adenosine triphosphate (ATP) levels were examined. The results showed that the learning and memory abilities and mitochondrial levels of ATP were significantly decreased in the D-gal-induced aging model group compared with those in the control group (P<0.01). In addition, marked changes in the mitochondrial functions and ultrastructure were observed between the groups. Seven weeks of L. plantarum NDC 75017 and D-gal coadministration significantly improved the learning and memory abilities of the rats compared with the D-gal-induced aging model group. Furthermore, the combination regime significantly improved the mitochondrial ultrastructure and functions, including the mitochondrial respiratory chain, mitochondrial membrane potential and mitochondrial permeability transition. The results revealed that the L. plantarum NDC 75017 was able to alleviate learning and memory injuries in aging rats by reducing the mitochondrial dysfunction induced by D-gal.

Treatment of D-galactose induced mouse aging with &lt;i&gt;Lycium barbarum&lt;/i&gt; polysaccharides and its mechanism study

We evaluated the effects of Lycium barbarum polysaccharides LBP) on D-galactose aging model mouse, and explored its possible mechanism. Kunming mice were randomly divided into the control group, the model group, the high-dose LBP group, and the low-dose LBP group. Except the control group, D-galactose was used for modelling. The drug was administrated when modelling. Mouse behavioural, learning and memory changes were observed, and the contents of lipid peroxidation (LPO), lipofuscin (LF) and monoamine oxidase B (MAO-B) in mouse brain tissue and the weight of immune organs were measured after 6 weeks. Compared with the control group, mouse weight gain in the model group reduced significantly. Compared with model group, after mice drank LBP, the times of electric shock was less than aging mice (in which, the high-dose LBP group, P<0.05), and electric shock incubation period was longer (P<0.01). On Day 45 after modelling and drug administration, the contents of LPO, LF and MAO-B in mouse brain tissue in the model group increased significantly, while those in the drug administration groups decreased significantly. The thymus index in the aging model group decreased significantly; the thymus index and the spleen index in the high-dose LBP group and the low-dose LBP group rebounded significantly (P<0.01). We concluded that LBP has an anti-aging effect on D-galactose induced aging model mouse, and its mechanism may be related with the alleviation of glucose metabolism disorder and the resistance of the generation of lipid peroxide and other substances, which damage cell membrane lipid.

Study on establishment of kidney deficient aging model and comparison with D-galactose induced aging model

To establish a kidney deficient aging model (KDAM), assess it in antioxidant capacity, HPAT axis function and bone metabolism, and compare with D-galactose aging model. Aging rat model was established by injecting D-galactose solution, meanwhile dexamethasone solution was injected to establish kidney deficient aging model. Then these models were evaluated by serum MDA (malondialdehyde) and GSH-Px (glutathione peroxidase), liver SOD (superoxide dismutase), adrenal, thymus and spleen index, CD4(+), CD8(+), and serum COR (cortisol), BGP (bone Gla-protein), plasma ACTH (adrenocorticotropic hormone) and CRH (corticotropin-releasing hormone). Compared with the normal group, the aging model group and the kidney deficient aging group showed significant decrease in liver SOD activity (P < 0.01 on average) and significant increase in serum MDA content (P < 0.01 on average) , and the kidney deficient aging group revealed remarkable decline in plasma ACTH content (P < 0.05). Compared with the normal group and the aging model group, the kidney deficient aging model group's weight, serum GSH-Px decreased (P < 0.01, P < 0.05), adrenal index decreased (P < 0.05, P < 0.01), serum COR decreased (P < 0.05 on average), plasma CRH increased (P < 0.05, P < 0.01), serum BGP content significantly decreased (P < 0.01 on average), value of CD4(+), CD8(+) decreased (P < 0.05, P < 0.01), CD4(+)/CD8(+) increased, but without significant difference. The kidney deficient aging model shows significant decrease in antioxidant capacity, dysfunction of HPAT axis disorder and abnormal bone metabolism. However, D-galactose aging model only shows a significant difference in antioxidant capacity.

Polysaccharides from the Medicinal Mushroom Cordyceps taii Show Antioxidant and Immunoenhancing Activities in a D-Galactose-Induced Aging Mouse Model.

Cordyceps taii, an edible medicinal mushroom native to south China, is recognized as an unparalleled resource of healthy foods and drug discovery. In the present study, the antioxidant pharmacological properties of C. taii were systematically investigated. In vitro assays revealed the scavenging activities of the aqueous extract and polysaccharides of C. taii against various free radicals, that is, 1,1-diphenyl-2-picrylhydrazyl radical, hydroxyl radical, and superoxide anion radical. The EC50 values for superoxide anion-free radical ranged from 2.04 mg/mL to 2.49 mg/mL, which was at least 2.6-fold stronger than that of antioxidant thiourea. The polysaccharides also significantly enhanced the antioxidant enzyme activities (superoxide dismutase, catalase, and glutathione peroxidase) and markedly decreased the malondialdehyde production of lipid peroxidation in a D-galactose-induced aging mouse model. Interestingly, the immune function of the administration group was significantly boosted compared with the D-galactose-induced aging model group. Therefore, the C. taii polysaccharides possessed potent antioxidant activity closely associated with immune function enhancement and free radical scavenging. These findings suggest that the polysaccharides are a promising source of natural antioxidants and antiaging drugs. Consequently, a preliminary chemical investigation was performed using gas chromatography-mass spectroscopy and revealed that the polysaccharides studied were mainly composed of glucose, mannose, and galactose. Fourier-transform infrared spectra also showed characteristic polysaccharide absorption bands.

Investigation of thrombospondin-1 and transforming growth factor-β expression in the heart of aging mice

Elderly patients face the problems of morbidity and mortality due to age-mediated disabilities. The purpose of the present study was to investigate the expression of thrombospondin-1 (TSP-1) and transforming growth factor-β (TGF-β) in aging mice, and its probable mechanism in the pathological changes of aging myocardium. The aging model group (AM) comprised 30-month-old mice, while the control group comprised 2-month-old mice. The pathological changes were explored by H&E staining, and the contents of superoxide dismulase (SOD) and malondialdehyde (MDA) in the hearts were determined by xanthine oxidation or TBA colorimetry. TSP-1 and TGF-β expression in the left ventricular myocardium was also measured by immunohistochemistry. The results showed that the activities of SOD decreased and the MDA content increased markedly in the hearts of the AM group compared to the control group. H&E staining showed that the control group myocardial cells lined up in order with clear structure and stained equably, while the AM group myocardial cells lined up in disorder with an augmented cell body and the appearance of many granules and interstitial fibrosis. Compared to the control group, in the hearts of the AM group, TSP-1 and TGF-β protein expression in myocardial cells showed a significant increase (P<0.01). TSP-1 and TGF-β expression increased in the myocardium, which may be related to pathological changes of age-related heart diseases, such as hypertrophy, fibrosis of myocardial cells and microvessel dissepiment thickening.

Retard of D-Galactose-Induced Aging in Mice by Whey Hydrolysates

The antioxidant capacities and anti-aging properties of whey hydrolysates fermented by Lactobacillus plantarum TUST200209 and its purified whey peptides were studied, which were dose-dependent and category-dependent. The treatment groups were orally administered high dose (1.5g/kg/day) and low dose (0.5/kg/day) of whey hydrosyates and its purified peptides, and high dose (1.5g/kg/day) of 0.9% saline (vehicle group and aged model group) and mixture of whey and Lactobacillus plantarum TUST200209 in senescence accelerated Kunming mice for six weeks, respectively. Whey hydrolysates and its purified peptides could increase levels of superoxide dismutase, glutathione peroxidase, and decrease contents of malondialdehyde in serum, quadriceps femoris and mitochondria, and for the high dose of whey peptides, most of the increase was significant (P＜0.05) compared with vehicle group. It demonstrated that the whey hydrolysates could significantly retard aging of the mice induced by free radical, especially for its purified peptides.

Changes of telomere and telomerase in effect of ginsenoside Rg1 to delay hematopoietic stem cell senescence

To investigate the roles of telomere and telomerase in the effect of ginsenoside Rg1 to delay hematopoietic stem cell senescence. Sca-1(+) HSC was isolated by magnetic cell sorting(MACS) and divided into five groups: the control group, the aged model group, the Rg1 group, the Rg1 treated aged group and the Rg1 delayed aged group. The changes of cells were observed by senescence-associated beta-Galactosidase (SA-beta-Gal) staining. Cell cycle assay and culture of mixed hematopoietic progenitor cell were used to investigate the effect of ginsenoside Rg1 to delay Sca-1(+) HSC senescence. Telomere length and telomerase activity were detected by southern blotting and TRAP-PCR-SYBR Green staining. Compared with aged model group, the percentage of positive cells expressed SA-beta-Gal and the number of cells entered G1 phase were decreased and the number of colony of mixed hematopoietic progenitor was increased. It showed markedly decreased in the shortening of telomere length and reinforcing in the telomerase activity to Rg1 treated aged group and Rg1 delayed aged group. The change of Rg1 delayed aged group was significantly higher than Rg1 treated aged group. Activation of telomerase and prolonging of telomere length might be involved in the process of ginsenoside Rg1 to delay and treat the senescence of Sca-1(+) HSC.

Steve bAccumulation of nerve growth factor and its receptors in the uterus and dorsal root ganglia in a mouse model of adenomyosis

BackgroundAdenomyosis is a common gynecological disease, which is accompanied by a series of immunological and neuroendocrinological changes. Nerve growth factor (NGF) plays a critical role in producing pain, neural plasticity, immunocyte aggregation and release of inflammatory factors. This study aimed to investigate the expression of NGF and its two receptors in uteri and dorsal root ganglia (DRG) in an adenomyosis mouse model, as well as their relationship with the severity of adenomyosis.MethodsForty newborn ICR mice were randomly divided into the adenomyosis model group and control group (n = 20 in each group). Mice in the adenomyosis model group were orally dosed with 2.7 μmol/kg tamoxifen on days 2-5 after birth. Experiments were conducted to identify the expression of NGF- beta and its receptors, tyrosine kinase receptor (trkA) and p75 neurotrophin receptor (p75NTR), in the uterus and DRG in four age groups (90+/-5 d, 140+/-5 d, 190+/-5 d and 240+/-5 d; n = 5 mice in each group) by western bolt, immunochemistry and real time reverse transcription-polymerase chain reaction.ResultsAdenomyosis, which became more serious as age increased, was successfully induced in dosed ICR mice. NGF-beta, trkA and p75NTR protein levels in the uterus and trkA mRNA levels in DRG were higher in the older aged adenomyosis model group than those in controls (190+/-5 d and 240+/-5 d groups, P < 0.05). The expression of NGF-beta and its receptors in the uterus increased gradually as age increased for adenomyosis mice (190+/-5 d and 240+/-5 d, P < 0.05, compared with 90+/-5 d) but it showed little change in control mice. The mRNA level of trkA in DRG also increased as age increased in the adenomyosis model group (190+/-5 d and 240+/-5 d, P < 0.05, compared with 90+/-5 d) but was unchanged in controls. The mRNA level of p75NTR in DRG was not different between the adenomyosis and control groups and was stable from young to old mice.ConclusionsNGF- beta can be used as an indicator for the severity of adenomyosis. The gradually increasing level of NGF- beta and its receptors while the disease becomes more severe suggests an effect of NGF- beta on pathogenic mechanisms of adenomyosis.

Angiogenesis of brain after ischemia/reperfusion injury of brain in aged rats and changes in expressions of basic fibroblast growth factor and transformation growth factor-β1

To elucidate the mechanism of angiogenesis after cerebral ischemia/reperfusion (I/R) in the aged rats by observing the changes in expressions of basic fibroblast growth factor (bFGF) and transformation growth factor-β1 (TGF-β1). Young Sprague-Dawley (SD) rats were classified into groups by random digits table, and aged SD rats were stratified by different body weight. Rats were randomly divided into groups of sham operation, ischemia (I) 3 hours, I/R 1, 3, 6, 12 days, with 6 rats in each group. Focal cerebral I/R model was reproduced by intraluminal filament technique. Microvessel density (MVD) of brain tissue, sum area of lumens were observed, and the expressions of bFGF protein, TGF-β1 protein and TGF-β1 mRNA were assessed with immunohistochemistry and hybridization in situ. MVD in young model group began to increase at I 3 hours, peaking at I/R 6 days, maintained up to I/R 12 days. MVD in aged model group began to descend at I 3 hours and continued to I/R 12 days. Sum area of lumens in young model group increased markedly at I/R 1 day, gradually lowered at I/R 1-6 days, and increased obviously again at I/R 12 days. Sum area of lumens in aged model group reached peak at I/R 1 day, gradually decreased subsequently. MVD in aged sham operation group were higher than that in young sham operation group (6.88±1.60 vs. 5.50±1.53, P<0.01). MVD and sum area of lumens in aged model group at I/R 1, 3, 6, 12 days were lower than young model group. Expressions of bFGF protein, TGF-β1 protein in young and aged model group were both gradually up-regulated, all of them reaching peak at I/R 3 days, and lowered gradually at I/R 3-12 days subsequently. Expressions of bFGF protein (grey level) in both aged sham operation group and those of model group at I 3 hours, I/R 1, 3, 12 days were lower than those of young sham operation and those of the model group at the same time points (176.80±5.10 vs. 172.82±1.53, 171.81±2.43 vs. 167.85±2.41, 167.99±5.51 vs. 164.90±2.15, 152.98±4.11 vs. 150.75±1.11, 165.67±3.55 vs. 161.73±1.29, P<0.05 or P<0.01). Expressions of TGF-β1 protein (grey level) in both aged sham operation and those of model group at I 3 hours, I/R 1, 3, 6, 12 days were all lower than those of young sham operation and those of model group at the same time points (182.69±3.12 vs. 176.13±4.08, 176.89±2.30 vs. 170.56±7.47, 171.74±2.70 vs. 165.43±2.91, 157.17±5.20 vs. 150.43±4.28, 161.72±4.81 vs. 155.37±2.92, 167.69±2.18 vs. 160.28±3.59, all P<0.01). TGF-β1 mRNA expressions in both young model group and aged model group reached peak at I/R 1 day, gradually lowered subsequently. Expressions of TGF-β1 mRNA (gray level) in both aged sham operation and those of model group at I 3 hours, I/R 3, 6, 12 days were lower than those of young sham operation and also model group at the same time points (176.51±9.52 vs. 169.09±5.08, 176.75±5.74 vs. 165.36±4.78, 177.33±5.68 vs. 165.25±8.14, 178.46±4.91 vs. 170.51±4.29, 203.95±4.51 vs. 181.98±5.59, all P<0.01). Angiogenesis is obviously weak after cerebral I/R in the aged, and the mechanism of which might be related to the down-regulation of expressions of bFGF protein, TGF-β1 protein and TGF-β1 mRNA. Aging factor may be one of the main reasons which induce the down regulation of expressions mentioned above.

A model of multiple organ injury induced by Klebsiella pneumoniae pneumonia in aged rats

To reproduce a model of bacterial multiple organ injury (MOI) in aged rats. Male Sprague-Dawley (SD) rats were used. The young rats were divided into young control group (YCG, n=10) and young model group (YMG, n=15), and the elderly, aged control group (ACG, n=10) and aged model group (AMG, n=25). The model of rats with Klebsiella pneumoniae pneumonia was produced by tracheal instillation of the bacteria, and injury to various organs was observed and evaluated with changes in biochemical parameters, pathological pictures and mortality. Between YMG and AMG, the mortality rates were 33.33% (5/15) and 60.00% (15/25), respectively, at 24 hours after instillation of the bacteria. Compared with YCG and ACG, the neutrophil percentage and white blood cell (WBC) counts in peripheral blood increased significantly in YMG and AMG groups (all P<0.01), the rates of dysfunction of the lungs, the heart and the liver, were 60%-100%. The respiratory dysfunction was evidenced by an increase in the arterial partial pressure of carbon dioxide (PaCO(2), P<0.01), and a decrease in the arterial partial pressure of oxygen (PaO(2), P<0.05 or P<0.01). Myocardial dysfunction was shown by a the sharp increase in creatine kinase (CK), creatine kinase isoenzyme MB (CK-MB) and lactate dehydrogenase (LDH), and that of the liver by changes in alanine aminotransferase (ALT) and aspartate aminotransferase (AST, P<0.05 or P<0.01). The pathological changes under light and electronic microscopy were obvious, and the main feature was infiltration of the inflammatory cells. Compared with YMG, PaO(2) in AMG dropped significantly, PaCO(2) increased, CK, CK-MB, LDH, ALT and AST also increased significantly (P<0.05 or P<0.01). The scores of pathological injury in the lungs, the heart and the small intestine in the AMG were obviously higher than that in YMG group (all P<0.05), and the same was trend in the liver and the kidney. The model of bacterial MOI in aged rats is reproduced successfully, and it mimics the pathogenesis of multiple organ dysfunction syndrome (MODS) which initiates from infection in the lungs. The model is simple and convenient to replicate with a high success rate. The MOI in the aged rats is characterized by the severity of the organ injury and a high mortality rate.

Effects of an aging vascular model on healthy and diseased hearts

The vitamin D(3) and nicotine (VDN) model is a model of isolated systolic hypertension (ISH) due to arterial calcification raising arterial stiffness and vascular impedance similar to an aged and stiffened arterial tree. We therefore analyzed the impact of this aging model on normal and diseased hearts with myocardial infarction (MI). Wistar rats were treated with VDN (n = 9), subjected to MI by coronary ligation (n = 10), or subjected to a combination of both MI and VDN treatment (VDN/MI, n = 14). A sham-treated group served as control (Ctrl, n = 10). Transthoracic echocardiography was performed every 2 wk, whereas invasive indexes were obtained at week 8 before death. Calcium, collagen, and protein contents were measured in the heart and the aorta. Systolic blood pressure, pulse pressure, thoracic aortic calcium, and end-systolic elastance as an index of myocardial contractility were highest in the aging model group compared with MI and Ctrl groups (P(VDN) < 0.05, 2-way ANOVA). Left ventricular wall stress and brain natriuretic peptide (P(VDNxMI) = not significant) were highest, while ejection fraction, stroke volume, and cardiac output were lowest in the combined group versus all other groups (P(VDNxMI) < 0.05). The combination of ISH due to this aging model and MI demonstrates significant alterations in cardiac function. This model mimics several clinical phenomena of cardiovascular aging and may thus serve to further study novel therapies.