| Dendrobium officinale Kimura et Migo, Orchidaceae, is a rare and endangered species in China, and a precious Chinese medicine with a long history. As a kind of important biological response modifiers, the previous research has proved polysaccharide was the major active constituents in Dendrobium. There are many published reports that have demonstrated polysaccharide has beneficial activities in antioxidant, immune stimulating and antitumor activities. Recently researches have proved that Dendrobium officinale (D. officinale) have effects of anti-oxidation in vitro, antitumor, immunoregulation, etc, so Dendrobium officinale are applied in clinic and Chinese herbal compound. However, research about anti-aging effects and related mechanism of Dendrobium officinale in vivo are still under investigation. This research used fresh Dendrobium officinale which was cultivated artificially,extracting its juice and then after separation and purification to get the Dendrobium officinale polysaccharides (DOP) which were substance we study on.We set off from the preparation of Dendrobium officinale tester. Applying fresh caudices Dendrobium officinale, after cleaning, cutting, extracting juice by using the juice extractor to get the Dendrobium officinale juice; Dendrobium officinale polysaccharide (DOP) was isolated from juice by ethanol precipitation. Then the extract was recovered by freeze-drying, which produced a crude polysaccharide. According to the Chinese Pharmacopeia 2010 version, we used the phenol-sulphuric acid assay to detect the amount of total polysaccharides, for the sake of dismissing the differences between different batches.Then, we conducted the general pharmacodynamics of Dendrobium officinale. Adopting dimethylbenzene-induced ear edema method, acetic acid-induced capillary permeability method and cotton ball granuloma rats model to observe the anti-inflammatory effect; using the swimming test to observe the anti-stress effect; Carbon clearance method was used to evaluate the effects on immune function; applying the ovariectomized mice models to obverse the anti-oxidation effect by detecting the SOD and MDA in blood serum and determining the thymus index and uterus index. We found that the extraction of Dendrobium officinale can inhibit the dimethylbenzene-induced ear edema in mice and the increased peritoneal capillary permeability in mice induced by acetic acid. And phagocytosis of phagocytes of mice was significantly improved and swimming time was prolonged by juice of Dendrobium officinale at high dose. High dose of Dendrobium officinale could increase the thymus and uterus indices, improve the SOD level and decrease the MDA level in blood serum of ovariectomized mice models. In this assay, the anti-inflammation effect and immuno-regulation effect of juice were better than the water extraction.After the general pharmacodynamics studies of Dendrobium officinale, we applied it on the aging models, including D-galactose induced sub-acute aging female mice model and the natural female aging mice model. We observed the anti-aging effects by praxiology research including step-down test and maze test, determining the viscera index. Besides, Pathological inspection was applied in important organs. In the sub-acute aging female mice model, we found that Dendrobium officinale can reduce the time of passing through the labyrinth and wrong times of D-galactose induced sub-acute aging mice model, augment the thymus and spleen indices. Pathological inspection had proved that it can improve the pathological change of uterus, ovary, and spleen. In the aging female mice, Dendrobium officinale also showed its way of anti-aging effects. It also reduced the time of passing through the labyrinth and wrong times. Ovary index, thymus index and spleen index are significantly increased by DOP on natural aging mice model, which also decreased the severity of swelling of uterus. Pathological inspection had proved that it can improve the pathological change of uterus, ovary, and spleen. Dendrobium officinale showed its the especial protection of immune organs and reproductive organs from the organ indices and pathological inspection In conclusion, Dendrobium officinale had the exact anti-aging effect on the whole on this two aging models, especially giving a great performance on enhancing learning and memory ability, increasing immunity and improve the pathological change of organs on a whole.By means of sub-acute aging model, anti-aging effect in vivo has been proved. We went further studies on the mechanism aging and the anti-aging effect of Dendrobium officinale. At first, determining the viscera index and SOD, GSH-Px in blood serum and important viscera base on the relationship of free radicals and aging. We found that SOD in serum, homogenate of heart, liver, kidney and GSH-Px, T-AOC in serum, CAT in heart were increased by Dendrobium officinale, which showed that it can increase the anti-oxidative enzymes and resist the attack of free radicals on organs. Moreover, to explore the mechanism of aging-inflammation-immunity, the amount of cytokine in blood serum was determined by cytometric bead array (CBA) assay and the expression of p-NFKBp65、 IκB-a of oven (female important organ) was checked out by western blotting. Five pro-inflammatory cytokines tested were strikingly reduced and the inhibited cytokine IL-10 was increased. Western blot results show that the expression of p-NFκBp65 increased in aging mice, and DOP can reduce this effect to a nearly normal situation. This experiment had demonstrated that aging was a chronic inflammation process and the inhibition of the development of inflammation could achieve the purpose of anti-aging. The anti-aging effect of DOP was better than juice, this is to say, DOP may be an inflammation inhibitor. Besides, in-vitro test was applied to reflect the anti-aging effect from a side view. Spleno-lymphocytes were cultivated in vitro to observe the effect of DOP on spleno-lymphocyte proliferation. This indicated that the outstanding immune stimulating effect of Dendrobium officinale lied in the physiological activity of polysaccharides.On one hand, DOP can increase the anti-oxidationase and promote the proliferation of spleno-lymphocytes; on the other hand, DOP can inhibit the release of pro-inflammatory cytokines and inhibit the NF-κB signal pathway to reach the anti-aging aim of anti-oxidation, immunoenhancement, and inhibition inflammation aging. |
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