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Aim To investigate the effect of Rhizoma Polygonati( RP) on the functional activity and the level of reactive oxygen species (ROS) on the subculture of endothelial progenitor cells ( EPCs) in aging rats. Methods EPCs from bone marrow in rats were isolated and cultured in vitro and identified. The 2nd EPCs were divided into fourgroups, which were subcultured to the 4th, 6th and 8th passage with application of drug - serum containing RP or not. The positive rate of cell senescence was detected by beta - galactosidase, the proliferation, migration and tubule formation were assayed by MIT, transwell chamber and in vivo angio- gcnesis kit, respectively, and ROS level was detected by flow cytometry. Results In the process of EPCs subculture, the positive rate of cell senescence gradually increased, accompanied by a significant decrease in proliferation, migration and tubule formation. The level of ROS increased sharply (P<0. 05). RP could reduce the positive rate of cell senescence, enhance the functional activities, and decrease the level of ROS of EPCs significantly (P < 0. 05). Conclusions RP may delay the aging process of EPCs in vitro and protect the function of EPCs by decreasing the level of ROS.
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To investigate the amelioration effect of saponins extracted from Panax japonicas (SPJ) on myocardial fibrosis in natural aging rats and its mechanisms, male SD rats aged 18 months were randomly divided into 3 groups (aging model group, low-dose SPJ group and high-dose SPJ group), with 10 rats in each group. SPJ groups were given SPJ at different doses (10, 60 mg·kg⁻¹·d⁻¹) consecutively for 6 months, meanwhile, aging model group was treated with the equal volume of saline for 6 months until 24 months old. Another 10 rats aged 6 month were used as young control group. The changes of myocardial morphological were observed by haematoxylin-eosin (HE) staining. Masson staining was used to observe the changes of collagen deposition in rat hearts. RT-PCR was used to detect the mRNA expression levels of myofibroblast marker α-SMA, collagen-related protein COL1α2, COL3α1 and matrix metalloproteinase MMP2, MMP9. Western blot was used to test the changes of the protein expressions of TGF-β1, p-Smad3, IL-1β and TNF-α in heart tissues. SPJ can effectively improve the arrangement of myocardial fibers, decrease inflammatory infiltration and reduce collagen deposition in aging rats. SPJ can effectively down-regulate the mRNA expression levels of COL1α2, COL3α1, α-SMA, MMP9, MMP2 and inhibit the protein expressions of TGF-β1, p-Smad3, TNF-α, IL-1β in the natural aging heart tissues. SPJ can effectively alleviate myocardial fibrosis in natural aging rats, and its mechanisms was related to the inhibition of the protein expressions of TGF-β1, p-Smad3 and the reduction of myocardial inflammation in rat hearts.
Subject(s)
Animals , Male , Rats , Fibrosis , Panax , Rats, Sprague-Dawley , Saponins , Signal Transduction , Smad3 Protein , Transforming Growth Factor beta1ABSTRACT
Objective To explore the effects of Guilingji on improving learning and memory dysfunction caused by aging. Methods The mouse model of subacute aging caused by D-galactose and the rat model of natural aging were used respectively to imitate learning and memory dysfunction caused by aging. The effects of Guilingji on improving of learning and memory function index were focused in the diving platform experiment and the Morris water maze experiment. At the same time, the effect of that on rat organ index and blood biochemical index were investigated. Results Guilingji can significantly prolong step down latency (P < 0.05) and reduce the number of errors within 5 min (P < 0.05, 0.01) of the model mice. It can shorten positioning navigation escape incubation period (P < 0.01), extend the space exploration quadrant retention time (P < 0.05, 0.01), and increase the number of access to the platform (P < 0.05). Guilingji can increase testicular, thymus and spleen index (P < 0.05) and reduce the ALT content in serum (P < 0.05). Conclusion Guilingji can obviously improve the impairment of learning and memory function caused by aging. It also has some good effects on enhancing immunity, improving reproductive capacity, and protecting liver.
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Objective To observe the effects of Bushen Huoxue Formula on free radical metabolism and p16 protein expression in heart of aged rats; To discuss its protective mechanism to the heart of aged rats.Methods One hundred were divided into young control group, the natural aging group,Bushen HuoxueFormula high-, medium- and low-dose groups, with 20 rats in each group. Each medication group was given relevant medicine for gavage, once a day for 16 weeks. 1 hour after the last administration, after the rats were sacrificed, serum and heart were taken. The contents of NO and MDA and activities of CAT and SOD in serum and myocardial,β-galactosidase enzyme and p16 protein expression in myocardial tissue were detected.ResultsCompared with the natural aging group, NO content and SOD activity in the serum of rats inBushen Huoxue Formula high-dose group increased significantly (P<0.05) and MDA content in allBushen Huoxue Formula groups decreased (P<0.01); NO content, CAT and SOD activity in the myocardial tissue of rats inBushen HuoxueFormula high-dose group increased significantly, and MDA content decreased significantly (P<0.05). CAT activity in allBushen HuoxueFormula groups increased (P<0.05,P<0.01).β-galactosidase enzyme and p16 protein expression in myocardial tissue in allBushen Huoxue Formula groups decreased.Conclusion Bushen Huoxue Formula can in hibit the aging of myocardial-aged rats, and the mechanism might be related to its anti-oxidative damage and inhibition of tumor suppressor gene p16 expression.
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To investigate the effects of saponins extracted from Panax japonicus(SPJ) on cardiomyocyte apoptosis in natural aging rats and explore its underlying mechanisms. SD male rats were randomly divided into four groups: young control group, natural aging group, SPJ low dose group and SPJ high dose group, with 10 rats in each group. The rats in natural aging group, SPJ low and high dose groups were respectively treated with normal saline, SPJ 10 and 60 mg•kg-1•d-1 from the beginning of 18 month-old, 6 days per week for 6 months till 24 month-old. Then the animals were sacrificed. Their myocardial morphology changes were observed by using haematoxylin-eoin(HE) staining; cardiomyocyte apoptosis was tested by using Tunel assays; and the protein expression levels of Bcl-2, Bax, IL-1β, TNF-α, AMPK, p-AMPK, Sirt1, and Ac-NF-κB p65 in myocardial tissues of rats were detected by Western blot. The results showed that SPJ could effectively improve the arrangement disorder of myocardial fibers, reduce the infiltration of inflammatory cells and inhibit cardiomyocyte apoptosis in natural aging rats. At the same time, SPJ could significantly inhibit the protein expression of Bax, IL-1β, TNF-α and Ac-NF-κB p65, and increase the expression of Bcl-2, Bcl-2/Bax, p-AMPK/AMPK and Sirt1 in the heart tissues of natural aging rats. SPJ can effectively inhibit cardiomyocyte apoptosis in natural aging rats, and its mechanisms may be related with the regulation of inflammatory reaction by AMPK/Sirt1/NF-κB signaling pathway.
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Objective To evaluate the role of simvastatin in preventing and curing osteoporosis vertebrae by examining the effect from simvastatin on osteogenesis of the lumbar vertebrae in aging rats. Methods Sixty 15-month-old male SD rats were divided into six groups: the control group (injected with normal saline for three month), the baseline group (executed upon the gastric irrigation), the simavastatin-treated group (gastric irrigation with simavastatin at the dose of 5, 10, 20 and 40 mg/kg/d, respectively, for three month). L4 vertebrae were checked by Dual-Energy X-ray Absorptionmetry (DXA), Peri-quantiy CT (pQCT) and mineral apposition rates test. L5 vertebrae were checked by mechanical compression test. Results The value of DXA, pQCT and mineral apposition rate of 10 mg simvastatin group were slightly higher than that of the control group, but no significant differences were found between the two groups. The bone material properties of 10 mg and 20 mg simvastatin group were better than those of the control group, with no significant differences. Conclusions Although 10 mg simvastatin group (equivalent to 12~24 mg/d for human) seemed to have better properties than the other simvastatin groups, but there were no significant differences among these simvastatin-treated group. It is indicated that simvastatin doesn't play a positive role in promoting osteogenesis of the lumbar vertebrae in aging rats, so it may have no preventing or curative effect for osteoporosis of the lumbar vertebrae.
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Objective: To investigate the effect of moxibustion on telomerase activity and genes expression in tissues of senescent rats. Methods: Subacute aging rats model were established by injection with D-gal solution. Points Shenshu (BL 23) were treated with moxibustion in treatment group, contrasting with a model group and a normal group. Enzyme-Linked Immunosorbent Assays(ELISA) was used for the level oftelomerase activity in liver tissues, and In Situ Hybridization(ISH) was used for the condition of expression of telomerase genes in liver tissues. Results: The level of telomerase activity in the aging model group was obviously lower than that in normal control group (P＜0.01), the level in moxibustion group was obviously higher than that in model group(P＜0.05). In comparison with normal rats, the positive-expressed areas and photodensity of telomerase genes in aging model group were all significantly lower (P＜0.01,P＜0.01), and the positive-expressed areas in moxibustion group was significantly higher than that in model group (P＜0.01). Conclusions: Moxibustion could regulate telomerase activity of senile rats, hence delaying aging.