Study On Antimutagenesis Of Aqueous Extract Of Pulsatilla Chinensis Regel

Nowadays,the accelerating industrialization proceeding and the contrivance and application of many new techniques increasingly change our material life,however,it also bring the severe hidden dan- ger to our lives as the aggravation of environmental contamination and the increase in mutagen.The mutagen can invade the body by many mays.It can induce gene mutation and chromosome aberranti- on,so that the body develop tumor,abnormal embryo,genetic defect and so on.So it is very important to search some thing which have the antimutagenesis from the environment to inhibit and degrade the high gene mutation rate and decrease the incidence of tumour and heredit- ary disease.China have abundant Chinese herbal medicine which is rich in biological active ingredient and less in toxic side effect and adverse effect.In the long term process of disease prevention and treating disease,It display a distinct ascendancy.It has been a hot spot to search the antimutagen from Chinese herbal medicine.Study on antimutagenesis of Chinese herbal medicine is an important part of Chinese medicinal materials research.In theprogress of research on antimutagenesis of Chinese herbal medici- ne,the main study both internal and abroad is concentrated on screening of the Chinese herbal medicine which have the effect of antimutagen,but the reports on definite mechanisms involved in antimutagenesis of Chinese herbal medicine have been hardly reported.Pulsatilla chinensis Regel has many pharmacological actions,such as heat-clearing and detoxicating,antiinflammatory,anti- causative organism,anti-parasite and so on,but the report of antimuta- genesis is scarce.This research is about the antimutagenesis and mechanism of Pulsatilla chinensis Regel.Taking CP as mutagen and Vc as control of antioxygen,we carried out a series of experiments on aqueous extract of Pulsatilla chinensis Regel as follows:lymphocyte proliferation, rate of micronucleus in mouse bone marrow PCE, activity of SOD and T-AOC in serum and the activity of NKc.The research offers the experimental evidence for the study on antimutag- enesis of Chinese herbal medicine,and it is valuable in theory and practice.The results were as follow.1. Definition of the best experiment dose of PWE. According to the clinical dose,we chose five different doses (10times,20 time,40 time,50 times and 60 times) about PWE,negative, and positive control group. Every group had eight mice.The route of administration was intragastric administration. Once a day .The negative,and positive control group were treated with partes aequales normal saline. The positive control and all of the different groups of PWE were treated with CP by intraperitoneal injection once daily for five days after continuous intragastric administration for six days group.The treatment continued for two weeks. Detected the stimulation index of lymphocyte and rate of micronucleus according to the methods of experiment two—experiment on lymphocyte transformation and experiment three—micronucleus test of bone marrow cells.From the results of stimulateon index of T lymphocyte and the rate of micronucleus,we concluded that 50 times was the best experiment dose of PWE in the range of dose chosen in the research after statistical analysis.2. Experiment on lymphocyte transformation. In the experiment,we set up four groups:positive control,PWE,P- WE+CP,and negative control.Every group had eight mice.Every mo- use was treated once a day for 14 days consistently by intragastric ad-ministration.The mice of positive control and PWE+CP were treated with CP by intraperitoneal injection once daily for five days after co- ntinuous intragastric administration for six days. Prepared the spleen suspension by sterile operation and carried out the experiment on ly- mphocyte transformation according to the MTT colorimetric method. The result indicated that PWE could enhance the function of both T and B lymphocytes.PWE not only could enhance the capability of T lymphocyte activated by ConA and the cellular immune function,but also could elevate the responsibility of spleen to LPS,incentive B ly- mphocyte multiplication,and enhance the humoral immune function, therefore antagonize the immunosuppression and genetic injury caus- ed by CP.3. Micronucleus test of bone marrow cells.The mice were the same as those used in the experiment two,then took bone marrow to smear, observed 1000 PCE every mouse and ca- lculated the rate of micronucleus.The result indicated that PWE could obviously reduce the rate of micronucleus in mouse bone marrow ce- lls induced by CP,inhibited mutagenesis of CP,and antaconized the genetic injury caused by CP.4. The determination of the activity of natural killer cell.The mice were the same as the experiment of B blastisation of lymphocyte.The prepation the of spleen cell was the same as the ex- periment two.There were three groups,including effector cell(spleen cell), target cell(YAC-1cell) and experiment (spleen cell and YAC-1 cell),100 microliter cell suspension was planted into the 96 well cul- ture plate in every group.The activity of natural killer cell was deter- mined according to the MTT colorimetric method. The results indic- ated that PWE could elevate the activity of NK obviously, oppose the decrease of NK caused by CP,and strength the nonspecific imm- unity function.5. Determination of the activity of SOD and the total antioxidati- ve ability in serum.In this experiment ,we set up six groups:positive control,PWE,PW- E+CP,Vc,Vc+CP and negative control.Vc is a powerful antioxidant, so we chose it as the control of antioxygion.The blood was collected from eyeball.The results indicated that PWE could elevate the activit- y of SOD obviously,eliminate free radical,interrupt the oxidizing rea- ction,increase the content of T-AOC in serum and strengthen the totalanti-oxidative ability so that the body can oppose the damage caused by free redical.Conclusion:By the study on these experiments:T,B lymphocyte transformati- on,micronucleus test of bone marrow PCE,the activity of natural kil- ler cell, the activity of SOD and the total anti-oxidative ability in ser- um.We can draw the conclusion that the PWE can oppose the genetic damage caused by antimutagen in many ways.The PWE has the anti- mutagenesis.The possible mechanisms involved in antimutagenesis of PWE through the reseach show as follow:1. PWE can inhibit mutagenesis of mutagen,diminish the damage of DNA and increase the reparation potentia.It may be play the role by protection and reinforcement on Ts cell,and the inhibition on mixed function oxidase.2. Enhance the function of specific cell immunity,humoral immunity and non-specific inherent immunity,Enhance the activity of NK cell, release many kinds of cell factors,participate immunological regulation of organism,mediate ADCC,dismiss the damaged cell,en- hance organism immune function to ectogenic mutagen obviouslyand protect the chromosome from damage caused by mutagen.3. It has the antioxidation and the function to eliminate free radicals,interrupt and exterminate oxidizing reaction of free redical,enhance the stability of biomembrane,strengthen the antioxidative ability,and oppose the damage caused by mutagen.