Camp Yinwei Prescription Having On The Aging Human Skin Fibroblasts Vitality Effect Experiment Research

ObjectiveTo observe effects of camp yinwei prescription on cultured human skin fibroblasts(HSF); and observed different concentrations camp yinwei prescription drugs oncompatibility containing serum with the control group to delay senility of human skinfibroblasts (HSF);discuss the possible mechanism lyceum.Methods1Cell culture: HSF cell was from KunMing cell bank,inoculate in the culture flaskand culture40generation in order to it’s preburning.2Preparation of drug-containing serum: The health and clean of60male SD rats(weight:300±50g) were randomly divided into four groups(camp yinwei prescriptionof contains low drug, camp yinwei prescription of contains middle drug, camp yinweiprescription of contains high drug and blank controller group),every group gastriclavage concentration for1.5g Kg、3g Kg、6g Kg.Blank controller group gastric lavagefor normal saling.Ones a day,eight days in all.gastric lavage eighth after.One hourlater,10%chloralic hydrastwo intraperitoneal injection hocus.sterile operation drawblood from heart,room temperature stalling two hours.2500rpm centrifuge for15minutes, take out blood serum,fiter in super clean bench.56℃inactivation30minutes,-80℃refrigerator preserve is drug-containing serum.separation campyinwei prescription of contains low drug serum, camp yinwei prescription of containsmiddle drug serum.Camp yinwei prescription of contains high drug serum and blankcontroller group serum. Respectively prepared containing10%serum DMEMcomplete medium.3Preparation of camp yinwei prescription: camp yinwei prescription consist ofgensing,astragalus mongholicus and so on.Density unit(crude drug is**g/ml).endconcentration is4g/ml.4Cell growth curve measuing method:cell dissociation sometimes it’s turned cellsuspension.Regulation cell concentration reach to2.5×10~4/ml,inoculate in24holetissue culture plate.After anchorage dependent,foster by medicated serum.Get3holecell every group,every day.Use the cell counting chamber count cell’s amount.Calculating average. Cell culture in three days, not counting of cells to change fluid.Counts for10days to cell culture for the horizontal axis of the days, cell count for the vertical axis, draw cell proliferation growth curve.5Cell proliferation activity detection: MTT method in detecting variousexperimental cell proliferation activity. Measure the OD value6P38JNK, and ERK1/2protein immune cell chemistry to detect: cells use0.25%trypsin digestion. Made from single-celled levitation liquid. With5x10~4a/holedensity in a vaccination in advance disinfection cover glass six holes in training board,at37℃,5%CO2hatch box after incubation24h,0.01mmol/L of PBS washed2times. Fixed agent with appropriate fixed cells to testing drugs for p38JNK, serumand ERK1/2, the influence of expression. Set up the negative control experiment, withno resistance.Results1Normal rats of serum of growth and proliferation activity HSF have no effect.Having WeiFang camp with containing the drug to the growth of serum HSF hasstimulative effect. And can effectively relieve the HSF aging process. Who, havinglow dose group of camp, compensation in camp who dose group of having high camp,who dose group of compared with normal control group. P <0.05.2Different concentration camp, having the WeiFang medicated serum all canincrease the activity of HSF proliferation, and normal control group were comparedsignificant difference (P <0.05); Different concentration camp who, having the serumcontaining medicine group for the protection of between HSF function high dosegroup was higher than the dosage group, is superior to the low dose group of dosagegroup.(P <0.05)3Three normal rats serum of growth and proliferation of HSF activity without theinfluence, and tire bovine serum cultivate no difference,(P>0.05)ConclusionsNormal rats serum of growth and proliferation of HSF activity without theinfluence, and tire bovine serum cultivate no difference, and can be used in the relatedresearch; Different concentration camp who, having the serum containing medicine toHSF all have certain effect of aging, and who, having high dose group of camp serumlevels of anti-aging medicine role, having better than in camp who dose group ofmedicine serum; Compensation in camp who dose group of serum containingmedicine of delay aging skin is better than having low dose group of camp guardmedicine serum, irrigation under physiological saline rats, and tire bovine serumgroup results no significant differences.