| Aging is a long and gradual process that the physiological functions decline gradually in living organisms and they will die at the end. In recent years, the problem of aging has been becoming increasingly serious and more and more people have focus on the research on alleviating aging and the mechanism of anti-aging. In order to reduce the oxidative damage to the organism of free radicals, a lot of synthetic antioxidants have been widely used. Tricholoma lobayense, a kind of nutrition-rich and valuable edible fungus distributed in tropical and subtropical regions, possesses many kinds of biological activities. In recent years, a lot of researches have shown that synthetic antioxidants should be restricted due to their potential hazards related to organism health. In view of the potent danger, it is becoming increasingly important to search safe and non-toxic antioxidants from natural products to substitute the synthetic antioxidants. The researchers in our laboratory have focused on Tricholoma lobayense polysaccharides and their various biological activities in recent years. In our previous study, Tricholoma lobayense polysaccharide TLH-3 possessed very excellent antioxidant activity in vitro experiment and its ability to scavenge the DPPH radical was comparable to that of Vc with IC50 value of 126 μg/mL. Tricholoma lobayense polysaccharide TLH-3 also can reduce the content of MDA. Besides, it possesses the ability to protect the HELF cells injured by tert-butylhydroperoxide (t-BHP). Tricholoma lobayense polysaccharides TLH and TLH-3 can improve the antioxidant capacity of aging mice induced by D-gal and retard the aging process highly effectively. Therefore, to investigate the anti-aging bioactivity of TLH and TLH-3 further, the present study was conducted to determine the anti-aging activity of Tricholoma lobayense polysaccharides in vitro by t-BHP induced HELF cells damage and in aged mice induced by D-gal. The research provides a theoretical basis for the natural and active polysaccharides development into potent and potential medicine and dietary supplement to decelerate the aging process of organism.1) The research on the protective effect of Tricholoma lobayense polysaccharide TLH-3 on HELF cells induced by t-BHP. In this research, MTT method was used to select the suitable concentrations of t-BHP and TLH-3 to established cell model and treat injured HELF cells respectively. For assessing whether the observed protective effect of Tricholoma lobayense polysaccharide TLH-3 was related to HELF cell apoptosis, the cells were stained (with Annexin V-FITC and PI) and subjected to be analyzed by flow cytometry. Besides, β-galactosidase staining was used to detect the protective effect of Tricholoma lobayense polysaccharide TLH-3 on HELF cells injured by t-BHP. The results showed that the viability of t-BHP-induced HELF cells increased after being treated by Tricholoma lobayense polysaccharide TLH-3. Compared with model cells, with the concentrations of TLH-3 being raised, the percent of viable population increased and the positive ratio of β-galactosidase staining decreased in experimental group.2) The research on expression of different kinds of protein extracted from t-BHP-induced HELF cells treated with or without Tricholoma lobayense polysaccharide TLH-3. In these studies, the results have shown that after being treated by different concentrations of Tricholoma lobayense polysaccharide TLH-3, the expression of bcl-2 increased compared with model group cells. But the expression of bax, p53 and caspase-3 of HELF cells decreased in varying degrees when t-BHP induced cells were treated with Tricholoma lobayense polysaccharide TLH-3. It can be concluded that Tricholoma lobayense polysaccharide TLH-3 can regulate cell life cycle and attenuate cell aging through regulating the expression of cell proteins.3) The research on the protective effect of Tricholoma lobayense polysaccharides TLH and TLH-3 on mice induced by D-gal. Mice were randomly divided into five groups of 10 each. The aging model of mice was established by subcutaneous injection of D-gal that was dissolved in 0.9% normal saline to the back at a dose rate of 120 mg/kg body weight/day (BW/day) for six weeks continuously. Then mice were treated by intragastric administration:normal group (not treated with D-gal,0.9% saline solution), model group (treated with D-gal,0.9% saline solution), positive control group (treated with D-gal, Vc 100 mg/kg BW/day) and experimental group (treated with D-gal, Tricholoma lobayense polysaccharides TLH and TLH-3 200 mg/kg BW/day respectively). The results showed that compared with model group, the content of IL-6 in blood serum decreased, the content of LPF in kidney, heart and blood serum decreased, the activity of SOD increased and the content of MDA dropped in kidney, the content of AGEs decreased in liver and blood serum, the activity of MAO in kidney and liver decreased significantly in experimental group that treated by Tricholoma lobayense polysaccharides TLH and TLH-3. The results of HE staining showed that TLH and TLH-3 can repair the pathological changes of kidney. The immunohistochemistry results showed that the expression of p53 decrease after the aging mice being treated by TLH and TLH-3. Therefore, Tricholoma lobayense polysaccharides TLH and TLH-3 could effectively attenuate the mice aging and improve some parameters related to aging, and the protective effect of Tricholoma lobayense polysaccharide TLH-3 is better than that of TLH.In this research, we investigate the anti-aging activity of Tricholoma lobayense polysaccharides in vitro and in vivo from free radical and immune theory of aging in many aspects. The study found that Tricholoma lobayense polysaccharides possessed excellent anti-aging activity both in vitro and in vivo, and can be developed into potent and potential medicine and dietary supplement to decelerate the aging process of organism. The study provides a theoretical basis for further research of natural polysaccharides on anti-aging. |
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