The Radioprotective Mechanism Of Fangfu Capsule Against Radiation Damage

With the modernization of science and technology, nuclear energy hasbeen widely used in all aspects of production, scientific research, medical,military, and also because of this, the increasing number of human exposure toradiation exposure. The same time as the ship’s cabin environment temperatureand humidity, low-dose radiation and other adverse effects of factors, lowdoses of ionizing radiation has become the main factors of the nuclearsubmarine long flight crew health damage tothe constraints of combat effectiveness. By traditional Chinese medicinetheory, rousing, virtual make up the governance guidance, screening todevelop anti-radiation capsule, the basis of modern pharmacology, to study theprotective effect in the blood and immune systems, while its antioxidantanalysis and DNA damage protection mechanism. Provide a theoretical basisfor future Chinese herbal compound in the in-depth study of DNA damagerecovery and immune.[Objective]Uniform design optimization the composition of Fangfu capsule. In theprescription optimized on the basis of Fangfu capsule pharmacodynamics andprotection mechanism. Maximum tolerated dose experiment to assess thesecurity of Fangfu capsule.[Methods]Prescription optimization The uniform design method, the five kinds of traditional Chinesemedicine in Fangfu capsule, divided into12groups according to the ratio ofadministration. Administered continuously for14days. In the first seven daysof the administration, the use of60Coγ of exposure to radiation damage model.And after irradiation3days,7days, blood detected by hemocytometer.Weighed everyday. Indicators above filter out the best prescription.Pharmacodynamic1．One hundred Kunming mice(half female, half-male), divided into fivegroups. Administration for14consecutive days, in the first seven days of theadministration, irradiation with60Coγ source radiation damage model.Weighed every day, and to observe the survival of irradiated mice for30d,record the state of the mouse and the number of deaths calculated30d survivalrate of mice, the rate of improvement and protection factor.2．Fifty male Kunming mice were randomly divided into five groups.Administration for14consecutive days, in the first seven days of theadministration, irradiation with60Coγ source radiation damage model.Weighed every day.2hours after the last administration, eyeball blood, cutsto remove the spleen, thymus, and right femur.MechanismForty male Kunming mice were randomly divided into five groups.Administration for14consecutive days, in the first seven days of theadministration, irradiation with60Coγ source radiation damage model. Afterirradiation weighed every day, two hours after the last administration,complete the following indicators:Bone marrow DNA contentBone marrow micronucleus ratePeripheral blood comet assayPCR method of DNA-PKcs detection The Western bloting method detection of apoptotic genes and P53Flow cytometry apoptosis of spleen cells with cycleElisa method spleen, thymus caspase-3, caspase-9were detectedMaximum tolerated dose experimentForty Kunming mice, male and female, were randomly divided intonormal group, the administration group, n=20. Laboratory animals afterfasting for12h, the administration group with the highest drug concentrationand animal stomachcapacity gavage, oral dose of360times for a maximumtolerated dose experiment.2times/d administered normal group fed the samevolume of double-distilled water.Experimental animals after administration ofconventional farming, and observed for14consecutive days.[Results]Prescription optimization1. Effect of the weight of irradiated mice with10kinds prescriptionTwelve groups of initial body weight of mice were not statisticallydifferent.14days after the last weight,12groups of mice were growth.Compared with the irradiated control group, the growth of a party to thenormal group and5is the most significant statistical difference (P<0.01,P<0.05)., The5th set of square has a significant protective effect onradiation-induced weight loss in mice.2. Effect of blood system of irradiated mice with10kinds prescriptionThree days and7days after irradiation, compared with the irradiatedcontrol group,3days after irradiation, the normal group and10prescription group RBC, WBC recovery were significant differences(P<0.01), of which only5groupsside of the HGB improve significantlydifferent (P <0.01). On the7th day after irradiation,10set of square groupof RBC, WBC, HGB are recovered, there is a significant difference(P<0.01). Capsule prescription anti-radiation protective effect of radiation damage, the5th set of square mouse blood recovery with normal group, thesmallest difference, the protective effect is most obvious.Pharmacodynamic1. The influence of the Fangfu capsule for30d survival of irradiated mice, therate of improvement and protection factorCompared with normal group, after irradiation all other group’s survivaltime were decreased. Survival time of mice to illuminate in the control groupwere increased, medium dose group and high dose group of growth havesignificant difference (P<0.01).30d survival rate of groups of micerespectively: normal group was100%, the exposure control group is20%, lowdose group was45%, the medium dosage group95%, high dose group was95%. Thus we could see that radiation capsule can not only greatly improvethe lethal dose of60Coγ irradiation in mice survival, but also significantlyincrease the survival rate of mice and protect coefficient, it shows that thecapsule has a strong radiation effect.2. The protective effect of blood system with Fangfu capsule:Compared with normal group, in addition to illuminate control group andmiddle dose group, other groups have significant difference (P<0.01).Compared with irradiation in the control group, normal group and the threedose groups of RBC, WBC, PLT and BMNC were recovered, there wassignificant difference (P<0.01). Lavage of dose and high dose Fangfu capsulein mice spleen index has increased significantly (P<0.01). Pathologicalobservation showed that normal mice spleen smooth surface without splenicnodule formation; Gamma irradiation, atrophy of spleen in mice spleennodules protrusions, surface all is white, can find plenty of nuclear pyknoticcells under a microscope, a reduction in the normal cells, dual-core abnormalcells increased. But lavage radiation capsule in mice after irradiationirradiation splenic nodule number generated significantly than the control group, a significant statistical difference (P<0.01), of which the dose group ofsplenic nodule number generated the most, pathological and normal group, thedifference between the minimum.3. The protective effect of immune system with Fangfu capsule:Compared with normal group, after irradiation, spleen T and Blymphocyte proliferation response capability and CD4~+, CD8~+activity weresignificantly decreased, compared with the irradiated control group, capsulesmice fed different concentrations of anti-radiation of T, B lymphocyteproliferation were restored, CD4~+, CD8~+increased activity, including therecovery of the middle dose group and high dose group, a statisticallysignificant difference (P<0.01, P<0.05). Serum after administration ofTGF-beta1, IL-2, IL-10are different, medium-dose and high-dose group hada statistically significant difference (P<0.01, P<0.05).Mechanism1. The protective effect of DNA damage with Fangfu capsule:Comet assay and bone marrow DNA content experiments have shownthat ionizing radiation on DNA damage is very obvious, there is asignificant difference (P<0.01) compared with normal group irradiatedcontrol group.Obtained by flow cytometry: irradiated spleen cellssignificantly increased apoptosis and cell cycle was arrested in G0/G1phase.Significant difference (P<0.01, P<0.05) compared with the irradiatedcontrol group, the administration group of mice was restored, which themiddle dose group. By PCR analysis found that the normal group andmiddle dose group, DNA-PKcs expression was significantly higher thanthat of the irradiated control group (P<0.05). Western blot analysis showed:compared with the irradiated control group, the administration group P53and decreased the expression of Bax, Bcl-2expression increased with doselinear relationship. 2. The protective effect of Oxidative damage with Fangfu capsule:Compared with normal group, the the irradiated mice peripheral bloodserum SOD and GSH-PX activity was significantly reduced MDA contentincreased significantly.Compared with the irradiated control group,administered mice peripheral blood serum SOD activity and MDA contentand GSH-PX activity were statistically significant differences (P<0.01).Maximum tolerated dose experimentResults show that no animal died, after feeding animal activity reducedslightly, there aren’t other abnormal biological characteristics, generallyreturned to normal within1h. Within2-14d behavior mental state is normal,bright color, feeding and drinking water in good condition, no adverse reactionoccurred. Put all the mice to death after14d, after dissecting autopsy we foundno abnormalities of the internal organs. there was no statistically significantfor the comparision of2set of changes in body mass in mice (P>0.05).