| Trollius chinensis belonging to dry flower and bud of buttercuptrollius chinensis, which is a kind of herbaceous perennial. it is distributedmostly in north and southwest region of China. The flower has a longhistory in China as a drug, it is used to remedy disease in folk, such asamygdalitis, mouth throat swollen, chordapsus and so on. Modernpharmacology reseaches shows that trollius chinensis has manypharmacological activity, for example, bacteriostat, antiviral, antioxidationand so on, flavonoids whose effective component monomer are orientinand vitexin belonging to glycoside are stable activity component in trolliuschinensis. Now, domestic and overseas reseachers mainly focus onbacteriostat, anti-inflammatory, antiviral of aspects of flavonoids aboutthe pharmacological reseaches of effective component from trolliuschinensis. The reseach team studies external antioxidation of flavonoids,orientin and vitexin in trollius chinensis. The results shows thatflavonoids, orientin and vitexin of trollius chinensis have somefunctions ,such as, the elimination of superoxide, hydroxyl radical andDPPH, the protection of red blood cell hemolysis, proves that orientinand vitexin of effective elements of trollius chinensis have a externalantioxidation, providing experimental basis to further reseaches aboutinternal antioxidation. The subject compares orientin to vitexinantioxidation by studying internal antioxidation of orientin and vitexinform trollius chinensis on mice aged by D-galactose, and picks out thebest drug's structure activity relationship about the function of anti-aging,and provides the reseach direction for discovering and combining new typeanti-aging drug.The replication of aging model Use sub-acute aging method of D-galactose, to adjust drug doseaccording to weight, injecting D-galactose by enterocoelia everyday,giving drug continuely eight weeks. Take blood respectively from normalmice (injected saline according to weight) group and model mice groupwhen create model six weeks, detecting T-AOC, SOD, MDA. Getblood again from model group, detecting above indicators for checkingaging situation of mice when create model eight weeks.2 Experimental category and giving drugExperiment is divided into nine groups after create model successfully:model group, normal matched group, high, middle, low of dose group oforientin and vitexin, vitamin E matched group. Each treated groups aregiven drug by mouth irrigation stomach every morning from ninth weekand normal matched group and model matched group are given samevolume saline one time everyday. To sure low dose group 10mg/kg, middledose group 20mg/kg and high dose group 40mg/kg of orientin and vitexineveryday, vitamine E group 20mg/kg everyday. Weight one time everydayand adjust dose according to mice's weight at every turn, giving drug timeis eight weeks.3 Detection of mice general conditionDetect each group mice general condition, such as the situation ofingestion food and water, weight, range of motion, fur colour, glossinesschange and so on. Fast to fetch complete brain and weight it accurately andcheck the mass change of brain tissue of each dose group after cutting headup.4 The detection method of serum antioxidation enzymesCut head down and get blood, centrifuge blood for serum, checkT-AOC, CAT, SOD, GSH-PH.5 The detection method of antioxidation enzymes from tissue Fetch quickly liver, brain and kidney, make these into homogenate atfrozen condition after cutting head down, check CAT, SOD, GSH-PX,MDA from each tissue.6 The detection of the function of cell membrane transport ion of liver,brain, kidney from mice.Manipulate according to the explain of kit, compute enzymes'activityaccording to formula ATP enzyme of Sodium and potassium.7 The detection of lipofuscin from brain and the observe of morphology ofbrain tissue.7.1 The detection of lipofuscinPick mice brain off quickly, add to chloroform : methanol (V/V=2:1)and make it into tissue homogenate, then centrifuge, detect fluorescenceintensity ( exciting light wavelength is 360nm, launch wavelength is 420nm)from getting supernate. Compute the content of lipofuscin of brain tissuewith the fluorescence intensity 50 of new quinine sultate standard liquid.7.2 The detection of morphology of brain tissueDetect shape, size and amount of red cell, cell whether complete ornot, neat or not, degeneration or not by electron microscope. Observe thesea horse ultrastructure of mice including cell membrane and cell nucleusare whether neat and clear or not and lipofuscin is whether deposit or not.Experimental results show: 1 the content of T-AOC, SOD, CAT of bloodserum of margin and model group are conducted respectively to mathstatistics, which all have significant difference (P<0.05) at sixth week, thecontent of T-AOC, SOD, CAT of blood serum of margin and modelgroup are conducted respectively to math statistics, which all havesignificant difference (P<0.05) at eighth week, manifesting copysuccessfully mice model of subacute aging caused by D-galactose.2 The general shape detection of mice: control group appear to smooth andthin fur, favorable spirit, flexible action. Model group appear to gloomyand coarse fur, have unhair phenomenon severely, thin body, weary spirit. Fur smooth degree and weight of each dose of orientin and vitexinall have some recovery comparing to model group, each dose of orientinand vitexin can delay mice's brain tissue wither aged by D-galactose,have a marked difference comparing to model group (P<0.05), there is nomarked difference between orientin and vitexin to the same dose group.3 orientin and vitexin of trollius chinensis monomer can enhance theactivity of T-AOC, SOD, CAT, GSH-PX of mice's serum, each dose grouphave a marked statistics meanings comparing to model group (P<0.05,P<0.01). Middle and low dose groups of orientin of trollius chinensismonomer enharce more the activity of antioxidation enzymes from serumthan the same dose of vitexin, which have a statistics meanings (P<0.05).There is no marked difference comparing high dose group of orienti to thatof vitexin. Middle and low dose groups of orientin of trollius chinensismonomer enhance less the activity of antioxidation enzymes from serumthan vitamin E, which have a statistics meanings (P<0.05), There is nomarked difference comparing the high dose group of orientin of trollius tovitamin E.4 Orientin and vitexin of trollius chinensis can enhance the activity of SOD,CAT, GST-PX and can reduce the content of MDA all from liver, brain,kidney of mice, each dose group of orientin and vitexin have a statisticsmeanings (P<0.05, P<0.01) comparing to model group. Middle and lowdose groups of orientin of trollius chinensis monomer can enhance morethe activity of antioxidation enzymes from liver, brain, kidney of micethan the same dose of vitexin, which have a statistics meanings (P<0.05).There is no marked difference comparing high dose group of orienti to thatof vitexin. Middle and low dose groups of orientin and vitexin of trolliuschinensis monomer can enhance less the activity of antioxidation enzymesthan vitamin E, which have a statistics meanings (P<0.05). There is nomarked difference between high dose of orienti and that of vitexin. 5 Orientin and vitexin of trollius chinensis monomer can enhance theactivity of Na-K-ATP enzymes, Ca-Mg-ATP enzymes from liver, brain,kidney of mice, each dose group of orientin and vitexin have a statisticsmeanings (P<0.05, P<0.01) comparing to model group. Middle and lowdose groups of orientin of trollius chinensis monomer enharce more theactivity of Na-K-ATP enzymes, Ca-Mg-ATP enzymes from liver, brain,kidney of mice than the same dose of vitexin, which have a statisticsmeanings (P<0.05). There is no marked difference comparing high dosegroup of orienti to that of vitexin. Middle and low dose groups of orientinand vitexin of trollius chinensis monomer enharce less the activity ofNa-K-ATP enzymes, Ca-Mg-ATP enzymes from tissue than vitamin E,which have a statistics meanings (P<0.05). There is no marked differencecomparing high dose group of orienti to that of vitexin.6 Orientin and vitexin of trollius chinensis monomer can reduce the contentof lipofuscin from mice's brain tissue. Each dose group of orientin andvitexin have a statistics meanings (P<0.05, P<0.01) comparing to modelgroup. Middle and low dose groups of orientin of trollius chinensismonomer can reduce more than the same dose group of vitexin aboutdeceasing the content of lipofuscin from mice's brain tissue, which have astatistics meanings (P<0.05). There is no marked difference comparinghigh dose group of orienti to that of vitexin. Middle and low dose groups oforientin and vitexin of trollius chinensis monomer can reduce less thecontent of lipofuscin from brain tissue than vitamin E, which have astatistics meanings (P<0.05). There is no marked difference comparinghigh dose group of orienti to that of vitexin.7 The nerve cell amount of model mice's sea horse appear to be reducedapparently, cell rank is confused, there is cell bubble degeneration fromnerve cell of pallium under observing by light microscope. Each treat grouphave recovery over model group, especially recovery effect be appeared tothe high and middle dose group of orientin and vitexin. Under light microscope observing nerve cell of mice's sea horseregion discovery that model group appear to unclear and deformation cellnucleus's membrane of nerve cell, deposit excessively lipofuscin,mitochondria membrane was injured partly, a portion of endoplasmicreticulum appear to expand, dwell and bubble variant. Each treat grouphave recovery over model group, especially recovery effect be appeared tothe high and middle dose group of orientin and vitexin and vitamin E.The results show that Orientin and vitexin of trollius chinensismonomer can improve the general shape of mice, delay the wither ofmice's brain tissue, enhance the activity of T-AOC, SOD, CAT. GSH-PXfrom mice's serum, enhance the activity of SOD, CAT, GSH-PX,Na-K-ATP, Ca-Mg-ATP enzyme and reduce the content of MDA all comefrom the liver, brain, kidney of mice. It can reduce the content oflipofuscin from brain tissue of mice and improve the structure function ofsea horse nerve cell of aging mice. So Orientin and vitexin of trolliuschinensis monomer can delay apparently aging by whole adjust mechanismof a lot of way, tache and target point. Especially, the antioxidationfunction of the middle and low dose Orientin is over that of vitexin, thehigh dose group of Orientin and vitexin are equivalent to vitamin E infunction. |
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