| Tianmazhenxuan Granula was invented by professor Baotian Chen who is famous veteran teran doctors of TCM in southern medical university.It is composed of Decotion of Four Drugs,Ramulus cinnamomi,IndianBread,Oriental Waterplan Rhizome,Radix Glycyrrhizae,Largehead Atractylodes Rhizome and so on.It is used to treat vertigo in Chinese tradional medicine equal to Meniere disease in morden medicine.The basic pathological mechanism of Meniere disease is membranous labyrinth hydrops.It's clinical feature are paroxysmal vertigo,deafness and bombus.The firt age of onset aways between 30 and 50.Up to now,the cause of Meniere disease has no final conclusion.Many hakeem consider that it is probably connected with the disturbance of cochlea microcirculation,the disharmony of balance between production and absorption of internal lymph fluid,immune reaction and autoimmune abnormality,the disruption of membranous labyrinth and the disorder of autonomic nerve and so on.On account of the reason of Meniere disease is uncertain,the treatment of it is difficult usually.Professor Baotian Chen treated it with Tianma zhenxuan granula and acquired satisfactory effect frequently.Shu Huang,a doctor of southern reported that treat Meniere disease only with tianmazhenxuan granules could receive favorable effect.Zhiping Lv reported that he used tianmazhenxuan granules treat 38 patients with Meniere disease,the effective power was 96.7%The experience of professor Baotian Chen in treating Meniere disease is abundant.At present,the chinese patent medicine in therapy of Meniere disease are very few。So the request of a safety and effective medicine for the patient with Meniere disease is emergent.If tianmazhenxuan granules is exploited into a new drug,the advantage of society and economy will be enormous,so this is the objective of choosing this subject.Before exploiting into new drug,the preliminary assessment of safety and effective is absolutely necessary.It's validity has completed by doctorGuimei Cui.In this experiment we will execute the acute toxicity experiment and chronic toxicity experiment of Tianma zhenxuan granula.Partâ… The acute toxicity experiment of Tianma zhenxuan granula.Objective:To presume the acute toxicity level of Tianma zhenxuan granula by observing toxic symptom,toxic degree,toxic time of appearing and vanishing,the incidence rate of dead and calculating the maximum administration dosage after the last administration on experimental animal.Method:The LD50 of Tianma zhenxuan granula couldn't be calculated after preliminary test,so we just executed the maximum administration dosage test。40 mouses were divided into two groups randomly,there were the Tianma zhenxuan granula and the blank control group.There were 20 animal in each group and the numble of female and male were equal.After 12 hours absolute dieting but not inhibiting the water,the mouses in Tianma zhenxuan granula group were drenched 87.5%concentration,0.8ml/20g volume of Tianma zhenxuan granula solution by one time.The concentration and the volume all were the maximum.The mouses in blank control group were drenched the negative ion water of equal volume.In the day of administration,we observed the appearance,activity,respiration,excretion, defecation,fatality and so on of the mouses in each group strictly.The observation should continue 7 days.In the 7 clays we raised the animals conventionly,weighed them each day and recorded the fatality and symptoms of toxic or side effect.In the end of experiment,we sacrificed and dissected all the animals,then we examined the pathological changes of the main organs macroscopicly and calculated the maximally tolerate dose(MTD).Result:There were no abnormally seen in appearance,activity,respiration, excretion,defecation of the mouses in each group and none of the mouses was died.Before administration the body weight(g) of Tianma zhenxuan granula group and blank control group were correspondly 26.05±3.22,24.50±2.76 respectively,the eighth day after administration the body weight(g) of Tianma zhenxuan granula group and blank control group were correspondly 30.45±4.40,27.60±3.03 respectively,and there were no significant difference in group comparison.There were no obvious abnormality when we observed the main organs just as cardia,liver,spleen,lung, kidney,stomach intestine and so on by naked eye.We measured MTD≥35g/kg,the doage is 700 times more than clinical everyday oral dose of adult which is 0.05g/kg.Partâ…¡The chronic toxic experiment of Tianma zhenxuan granulaObjective:To observe the toxic reaction of Tianma zhenxuan granula in rats by successive and repeated administration.So we can find the potential adverse effect to hint the security index which should monitor intensively in clinical experiment and we can provide reference information to saving measure of clinical toxic reaction.Method:The number of wistar rats were 80,they were divided into four groups randomly.The four group were Tianma zhenxuan granula high-dose group,middle-dose group,low-dose group respectively.There were 20 rats in each group,the number of female and male were equal.We raised the rats for one week before experiment for adaptive observation.The administration dosage of high-dose group,middle-dose group,low-dose group were 2.7 g/kg,1.35 g/kg,0.68g/kg respectively.Those dosages of each group were 54 times,27 times,13.5 times respectively more than clinical everyday oral dose of adult which is 0.05g/kg.The administration volum of all rats in each group were 3ml/kg.We administrated the rats everyday morning for 12 weeks and weighed their body weights every week.In the experimental period of time we observed the appearance,activity,excretion,fatality and toxic reaction of each rat and so on.24 hours after the last administration,we executed 5 female and 5 male rats in each group to detect hemology index and blood biochemics index then we put them into system dissection,weighed the weight of the main organs and calculated the organ coeffeicent.At last,we made the pathological section of each organs and observed the patholagical changes of them through light microscope.After two weeks'observation and raise,the left rats of each groups were executed and the procedure was the same as above.Results:1 The influence of Tianmazhenxuan Granula on rats' general statesDuring the administrion and recovery period,there was no rat died in each group.The appearance,activity,respiration,excretion,defecation and so on of the rats in each group strictly were not observed abnormal.In the 5th week of administration period,there were two female rats depilated on their head,nape,back,periphery of fossa orbitalis and fore-leg.The color pattern were bright and the skin of depilation had no abnormality seen in color and texture.2 weeks later the depilating phenomenon recovered.There were no abnormal seen in other group.2 The influence of Tianma zhenxuan granula on rats' body weightsBoth the female rats' body weights(F=3.601,P＜0.05) and the male rats' body weight(F=1279.107,P＜0.001) of each group were in the increase in the administration period.But the incresing velocity of male rats'body weights was disparity(F=1791.081,P＜0.001),the increase of body weights in high-dose group and middle-dose group slowed down with the lasting of administration time and it showed time-effect relationship.There was no obvious time-effect relationship in low-dose group.There was reciprocation effect between administration time and administration dosage(F=2.477,P＜0.001).In administration period,there was no statistical significance in the body weights'differents of female rats in each group(F= 0.727,P=0.543),so did each time.There wasn't reciprocation effect between group and administration time(F=1.540,P=0.163).In the recovery phase,the body weights of male rats in each group increased with time(F=5.864,P＜0.05).The incresing velocity of male rats'body weights was at equal pace(F=1.759,P=0.195).There wasn't reciprocation effect between group and administration time(F=0.638,P=0.062).In the recovery phase,the body weights of female rats in each group increased with time except the middle-dose group(t=0.619, P=0.420).The body weight of each group was disparity(F=4.419,P=0.019),the body weights' variance of each group in 14th week had statistical significance (F=5.511,P=0.007).The multiple comparison between each group manifested that the high-dose group,middle-dose group and blank control group all lower than low-dose group.There wasn't reciprocation effect between group and administration time(F=2.463,P=0.100).3 The influence of Tianma zhenxuan granula on rats' hemology indexes.In the administration period,the difference of PLT in each group had statistical significance(F=2.962,P=0.045).Compared with blank control group,there were different increase in degree in other groups,the low-dose group was higher than blank control group obviously.The difference of WBC in each group had statistical significance(F=3.366,P=0.029).The high-dose group,middle-dose group and blank control group were lower than the low-dose group.The difference of LYM in each group had statistical significance(F=4.400,P=0.010).The high-dose group,middle-dose group and blank control group were lower than the low-dose group.The difference of MID in each group had statistical significance (F=3.065,P=0.041).The high-dose group,middle-dose group and low-dose group were higher than blank control group obviously.The difference of MID in each group had statistical significance(F=3.589,P=0.037),but the comparison between each two group had no statistical significance.In the recovery phase,all of the hemology indexes'difference in each group had no statistical significance,they all in the same level.4 The influence of Tianma zhenxuan granula on rats'blood biochemics indexes.In the administration period,the difference of BUN in each group had statistical significance(F=3.611,P=0.034),the middle-dose group was lower than low-dose group obviously and there had no significant dosage- effect relationship.The rest blood biochemics indexes'difference of each group had no statistical significance, they all in the same level. In the recovery phase,the difference of TG in each group had statistical significance(F=3.675,P=0.033),but the comparison between each two group had no statistical significance.The rest blood biochemics indexes'difference of each group had no statistical significance,they all in the same level.5 The influence of Tianma zhenxuan granula on rats'organ weight and organ coefficientIn the administration period,the differences of spleen coefficient and spleen humid weight in each group had statistical significance(the F values were 2.57,2.900 respectively,the P values were 0.046,0.049 respectively),the high-dose group and the blank control group were lower than middle-dose group.The difference of lung humid weight had statistical significance(F=3.281,P=0.032),the high-dose group,low-dose group and blank control group are lower than the middle-dose group.In the recovery phase,the difference of thoracic gland coefficient and thoracic gland humid weight had statistical significance(the F values were 3.689,4.177 respectively,the P values were 0.021,0.012 respectively),the middle-dose group were higher than blank control group.The difference of didymus humid weight in each group had statistical significance(P=3.485,F=0.041),the high-dose group and low-dose group were lower than middle-dose group.The difference of liver coefficient in each group had statistical significance(P=3.960,F=0.024),the low-dose group was lower than blank control group.6 The results of pathologic histology tested under light microscope.In the administration period,both in the high-dose group and the blank control group,there were two rats existed fat denaturation,acidophilia denaturation,swelling in the liver cell.Each of them had one or two pathological change.There were one or two rats exist interstitial pneumonia or chronic bronchitis.In the recovery phase,both in the high-dose group and the blank control group, there were two rats existed fat denaturation,acidophilia denaturation,swelling in the liver cell.Each of them had one or two pathological change.There were one or two rats exist interstitial pneumonia or chronic bronchitis.Conclusion1 In the acute toxic experiment,the MTD of mouses on Tianmazhenxuan granula≥35g/kg,the doage is 700 times more than clinical everyday oral dose of adult which is 0.05g/kg.2 Tianmazhenxuan granula promote the increasing of rat's body weight.3 The influence of Tianmazhenxuan granula on rats'hemology indexes was reversible and had no delayed reaction.4 Tianmazhenxuan granula had no obvious influence on rats' hemology indexes.5 Tianmazhenxuan granula had no obvious influence on rats'organ humid weight and organ coefficient.6 Tianmazhenxuan granula had no obvious influence on rats'pathohistology.7 Tianmazhenxuan granula has-no obvious toxic and side effects,it is safe for continuous oral administration. |
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