| 1 the reason to select topic and the aim to researchWith the aging problem of population looming large, it has become a big task of medical research how to delay senescence and to improve the quality life of the aged people. In recent years, based on the TCM theory of "foundation of the congenital constitution", a lot research work has been done, which shed some light on the mechanism of anti-aging with TCM. Nevertheless, aging is complicated process that involves in a comprehensive action of multiple factors, in which influence that the spleen acts as "foundation of acquired constitution" dose not allow to be ignored.The outline cognition of TCM to senility could go back to the age of "yellow emperor's internal classic". The book had a fairly all round discuss over the aging cause and its mechanism at a certain age. "internal classic" says: "by the age of seven for girls, and the baby teeth are replaced by permanent teeth, and hair grow longer and faster becauseof sufficient kidney Qi.......at the age of 35 yangming meridian declines, the face begins towither and hair to lose. ......by the age of eight for boys, and the baby teeth are replaced bypermanent teeth, and hair grow longer and faster because of sthenia nephropathy. ......atthe age of 64, the teeth and hair will lose. The kidney dominates water metabolism, which can exert the storing function by receiving vital essence from the viscera, so the kidney can discharge due to plenty of vital essence in the five viscera circulating normally. Now that all the viscera decline in function, and bones and muscles are soft and slack, and the kidney-essence to promote reproduce uses up, so hair on the temples is white, and the body weight is put on with staggering steps and without the ability to reproduce. "Plain question" states: by the age of ten, the functions of the five viscera begins stable, Qi and blood cancirculate around, their Qi locates below, so children love walking. ......at the age ofseventy, the spleen Qi is deficient, so the skin gets drying. At the age of one hundred, all the five viscera are deficient, and one's vigor and energy are already weak, and finally he will die with only the human body empty of them. As "spirit pivot" holds clearly that senility must happen with age, which is based on spontaneous physiologic and pathologic changes of viscera, Qi and blood.The theory of relation between aging and deficiency of the spleen and stomachoriginated from "internal classic" and was to some degree developed by doctors from the Tang, Song, Yuan, Ming and Qing respectively. The famous doctor with invigorating the spleen, Li Dongyuan, who put stress on the function of the spleen and stomach and drew it into the view of anti-aging, stated that although people, who were ill with the spleen and stomach, no longer suffered from other diseases, they had lost their age. He advocated that regulating the spleen and stomach should be done first for aged people ("the theory of the spleen and stomach"). Li Zhongzhi put forward that the congenital foundation rooted in the kidney, while the foundation of acquired constitutions was from the spleen. Zhang Jiebing upheld that "the spleen and stomach is the foundation of viscera", and earth is theorigin of all things. Stomach Qi is the foundation of health cultivation, ......so experts ofhealth cultivation are certain to put the spleen and stomach ahead, he, who is good atcultivating the spleen and stomach, can prolong his life("jingyue omni-books"). ......WuKeng stated: "all the viscera and body get Qi from the spleen and stomach first, and then become strong. If the spleen and stomach got weak, all the viscera and body couldn't get Qi and would gradually become weak" ("research on therapeutic formula"). Gong Yanting said: "all the aged people should stress on cultivating their primordial Qi and invigorating the spleen and stomach" ("discussion about senility"). Wang Wenlu held: "to support the spleen and stomach is the most important for health cultivation"("medical ancestors" ) and so on. In a word, much attention is widely paid to the spleen and stomach, which has been enormously woven into the theory and treatment of anti-aging, and plays the same role as anti-aging through invigorating the kidney and is regarded as the two most important principles of anti-aging.The past research has showed that to invigorate the spleen can be against aging through different links, in that how its anti-aging works is well worth being further researched. This paper will observe the influence of ShiJunZi(SJZ), a typical formula of invigorating the spleen to increase Qi, on the defect of mtDNA in the aging model of the mouse, in the hope to discuss over the relation between the spleen and aging at the genetic level. The findings will enrich the theoretic meaning of "spleen and stomach doctrine", afford new thoughts and the theoretic basis for anti-aging and establish the theoretic foundation for developing new anti-aging drugs.2 research methods2 1 establishment of mouse aging modelKuming mice of two month were randomly divided into six groups: normal control, model control, Vit-E (250mg- kg"1^"1) , SJZ high dose(20g-kg"1-d"1), middle(10g-kg"'-d"') and low (5g-kg"'-d"') groups. Aging model of mice was caused by D-galactose, water maze was used to testify whether the establishment of the model with learning disorder and dysmnesia is successful or not. 2 2 observation targets2 2 1 observation of ethology to observe the general condition and to test learning andmemory abilities in mice before and after modeling.2 2 2 evaluation of NO and NOS in the brain tissue of the mice nitrate reductase methodfor NO;to evaluate the activity of NOS according to extinction value of standardpreparation.2 2 3 activity evaluation of GSH-Px xanthine oxidasen method to measure the activity ofSOD, DTNB coloration for GSH-Px.2 2 4 specific activity of succinate dehydrogenase to perform according to kit instruction.2 2 5 defect measurement of mitochondria DNA in hippocampus tissue of the mice PCRamplification.2 2 6 calcium concentration evaluated by di-wavelength fluorometric method inhippocampus tissue of the mice fluorospectrophotometer method.2 2 7 sample preparation of TEM and its observation morphosis change of mitochondriawas observed with TEM and pictures were taken.2 2 8 apoptosis of brain cells flow cytometry3 experimental results3 1 observation of general state of healthMost mice manifested as dark hair or alopecie, depression, poor appetite, kinesis retardation and less activity in the model group;a fraction of the mice appeared the symptoms such as incoordination, more spontaneous movement or self biting and so on. Similar symptoms like the model group were showed after about one month in the therapeutic group with drugs, but they were lighter and gradually relieved within two months. The mice in the blank control group had no symptoms like the above mentioned. 3 2 SJZ effect on the improvement of learning disorder and dysmnesia caused by D-gal in the miceWhen handled by D-gal in eight weeks, the abilities of learning and memory in the mice of the model group significantly reduced, manifested as that the time to swim the whole range at the learning and remembering stage extended obviously, and the times to enter the blind-end remarkably increased, and ompared with the normal group, the difference was significant PO.01), but treated with SJZ of high, middle and low doses, the same behaviour as above mentioned in the mice well improved(P<0.01, P <0.05). 3 3 effect of SJZ on NO content and the activity of NOS in the brain tissue of the mouse with learning disorder and dysmnesia caused by D-gal.When handled by D-gal in eight weeks, NO content and the activity of NOS tn the brain tissues of the model group were much higher than that in the normal control group( P <0.01), while they were significantly lowered in the brain tissues of modeling mice-handled by SJZ of high, middle and low doses (PO.01, PO.05) . 3 4 effect of SJZ on the activities of GSH-Px and SDH in the brain tissue of the j?uusewith learning disorder and dysmnesia caused by D-gal.When handled by D-gal in eight weeks, the activities of GSH-Px and SDH in the brain tissues of the model group were much lower than that in the normal control group( P <0.01), while they were significantly higher in the brain tissues of modeling mice handled by Vit E and SJZ of high, middle and low doses(P<0.01, (PO.05). 3 5 effect of SJZ on the intracellular Ca2+ of nerve cells in the hippocampus of the mice with learning disorder and dysmnesia caused by D-gal.Compared with the control group, the activity of intracellular Ca2+ in the brain tissue of the modeling mice significantly increased (P<0.01).In comparison with the model group, after intragastric administration with Vit E and SJZ of high, middle and low doses in the sixth week, the activity of intracellular Ca2+ in the brain tissue of the mice with different doses of SJZ remarkably decreased (PO.Ol).3 6 effect of SJZ on DNA mutation of injured mitochondria in nerve cells of the mouse hippocampusA strap of 0.47kb clip could be seen in all the specimen of the hippocampus, in that there was another about 0.63kb clip amplified in the mice of the model group (FigA) . This demonstrated that there existed a clip defect of about 3.87kb in mtDNA of the mouse hippocampus from the model group.3 7 electron microscopeUltramicrostructure of hippocampus nerve cells was normal in the normal control group.In the hippocampus nerve cells of the model group such changes could be seen as cytoplasm rarefaction, higher electron density of endochylema with clear nucleolus, indistinct structure in most cell organelles, more deposit of lipofuscin, fewer mitochondria with swelling, light matrix, vacuolization, even a lot deformation and defect of mitochondria cristae.In the hippocampus nerve cells of the therapeutic groups, the cell edge was sharp with integrated cell membrane, clear double-decker. Mitochondria distributed with a great quantity, which had regular shapes, clear mitochondria cristae. Besides increased ribosome in cytoplasm, the others were nearly normal.4 conclusionThe aqueous extracts of different doses from SJZ could effectively interfere in encephalic aging, reduce the DNA defect of mitochondria in the hippocampus and improve the abilities of learning and retention. The possible mechanism of interfering in encephalic aging is that SJZ could enhance the ability of antioxygen, regulate the disequilibrium between the oxidative damage and antioxygen system, reduce the calcium ion concentration, decrease the activation of NOS, arrest over produce of NO, lighten neuronic injuries caused by free radical, diminish the defect of mtDNA in the hippocampus, improve the energymetabolism of mitochondria and keep its function. Inferred from the above mentioned, SJZ could exert its anti-aging action through multi-ways, multi-links and doing the poly-targets.5 SummaryBy synthesis and analysis of literature from theories of anti-aging mechanism and experiments with TCM, this paper takes "internal classic" and the cognition of relation between the spleen and stomach and aging as basis, combines with the modern lifestyle of people, especially its influence of ill-healthy eating habits on the function of the spleen and stomach, on the internal metabolism and on the probability to accelerate aging and takes SJZ, a representative recipe to invigorate the spleen- to increase Qi, as a means, in the hope to discuss the relation between the spleen and aging and elucidate the molecular action mechanism of anti-aging with invigorating the spleen to increase Qi at the genie level and the different technique depth.Looked from the experiment results, the means of invigorating the spleen to increase Qi is surely to interfere in the encephalic aging, to reduce the DNA defect of mitochondria in the hippocampus and to improve the ability to learn and remember. That means that it has a certain action on anti-aging, which further illustrates that the relation between the spleen and stomach and aging can not be neglected.Integrated with the exiting experimental results, separated recipe research can be done in order further to affirm the best grouping (single herb in effect or group of parts in effect) and establish the foundation to develop the new drugs for anti-aging. In addition, the material basis of SJZ to improve the abilities of space learning and memory in the mouse is left to research deeply. Due to a lot factors to affect the aging, this paper undertakes some observation only from several aspects of it, and more researches are waiting to carry out.
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