Analysis And Prevention Of Acute Gastrointestinal Radiation Injury By Traditional Chinese Medicine

Objective:1.To explore the appropriate method of establishing an acute radiation injury model of gastrointestinal radiation injury,evaluate the characteristics and applicability of the model,and lay the foundation for the traditional Chinese medicine(TCM)prevention method of acute radiation injury of gastrointestinal tract in sudden nuclear exposure.2.To explore the low doses of radiation injury prevention methods of traditional Chinese medicine by observing the dynamic changes of living animals,organs and tissues in experimental animals and summarizing the characteristics of Chinese medicine syndromes and evolution rule of acute gastrointestinal radiation injury model.3.To observe and evaluate the effect of heat clearing and detoxification cooling blood method on preventing acute gastrointestinal radiation injury and characteristics of Chinese medicine syndromes evolution.4.To explore the prevention mechanism of heat clearing and detoxification cooling blood method on acute gastrointestinal radiation injury.Methods:1.Research on modeling method:The rats were irradiated with 60Co-gamma ray,and the degree of injury was evaluated according to the mortality,feeding,defecation,anatomic and histological changes of the irradiated rats.The best dose was determined by two steps.It is the first choice to determine the dose range of irradiation by means of literature study.After the first irradiation,the dose range is optimized,then the dose is divided into a second fine screen.,the first dose range of irradiation was 16Gy,12Gy,8Gy,each group had 20 rats(10 rats were taken to observe survival rate respectively).After irradiation,the body weight,food intake,defecation,mental state,activity,hair luster,anus bleed,anus bleeding,emaciation,death and so on were observed every day.10 rats in the rest of each irradiation group were killed in the abdominal aorta after anesthesia at 5d,the lungs,the liver,the spleen and the kidney were observed by the naked eyes and the pathological changes of the esophagus,stomach,jejunum,colon and rectum were observed under the light microscope.At 14d,the blank group and the rats who were used to observe the mortality were executed,and the methods of observation were the same as above.The second step dose screening range was selected according to the results of screening.The three groups were selected again.2.Pathogenesis and syndrome analysis:90 male SD rats were randomly divided into blank group(n=10)and model group(n=80),The model group was irradiated with a single dose of 60 Co-y radiation and the control group was not irradiated.The general condition of the rats,include body weight,food intake,defecation,activity,hair removal,anus bleeding,emaciation,death were observed daily.At 1d,4d,7d,10d,14d,after irradiation,10 rats were randomly selected from the irradiated group and taken blood to be killed after the anesthesia.The gross specimens of the lung,liver,spleen,kidney and stomach were observed by the naked eyes,and histopathological changes in the stomach,jejunum,colon and rectum was observed with the microscope.The characteristics of syndrome were analyzed by the theory and method of TCM diagnosis.3.Evaluation of clearing away heat,detoxicating and cooling blood method to prevent acute gastrointestinal radiation injury:170 male SD rats were randomly divided into control group(n=10),irradiation group(n=80)and Chinese medicine group(n=80).The irradiation group and Chinese medicine group were irradiated with a single dose of 60 Co-y radiation and the control group was not irradiated.Before irradiation,Chinese medicine group was given intragastric administration of 2ml decoction,1 times daily for 7 consecutive days,the concentration was 1.42g/ml;the control group and irradiation group were given equal volume of saline 10 ml/kg orally for 7 days;The general condition of the rats,include body weight,food intake,defecation,activity,hair removal,anus bleeding,emaciation,death were observed daily.At 1d,4d,7d,10d,14d,after irradiation,10 rats were randomly selected from the irradiation group and Chinese medicine group,then they were taken blood to be killed after the anesthesia.The gross specimens of the lung,liver,spleen,kidney and stomach were observed by the naked eyes,and histopathological changes in the stomach,jejunum,colon and rectum were observed with the microscope.Evaluation of the preventive effect of traditional Chinese Medicine.4.The mechanism of clearing away heat,detoxicating and cooling blood method for prevention of acute gastrointestinal radiation injury:170 male SD rats were randomly divided into control group(n=10),irradiation group(n=80)and Chinese medicine group(n=80).The irradiation group and Chinese medicine group were irradiated with a single dose of 60 Co-? radiation and the control group was not irradiated.Before irradiation,Chinese medicine group was given intragastric administration of 2ml decoction,1 times daily for 7 consecutive days,the concentration was 1.42g/ml;the control group and irradiation group were given equal volume of saline 10 ml/kg orally for 7 days.At 1d,4d,7d,10d,14d,after irradiation,10 rats were randomly selected from the irradiation group and Chinese medicine group,then they were taken blood to be killed after the anesthesia.Test the thrombus and elasticity,measured the concentration of serum TNF-alpha,interleukin-6,interleukin-1 beta,thromboxane B2 and 6-prostaglandin in serum by ELISA,calculated the value of TXB2/6-Keto-PGF1 alpha;and detected the content of NF-kappa BmRNA expression in the jejunum tissue by RT-PCR method.Results:1.Research on modeling method:first dose screening:rats in the 8Gy group showed signs of short term bowel movement,no water and blood stool,the body mass and food intake of rats decreased not obviously,and no death occurred.No erosion and necrosis,exfoliation and bleeding were found in the intestinal mucosa with the microscope.Rats in the 12Gy group showed signs of water samples,partial blood water,refusal to eat,body mass decreased obviously,and intestinal mucosa erosion,necrosis,partial exfoliation,multiple bleeding foci with the microscope,the survival rate of two weeks was 20%.Rats in the group 16Gy showed signs of blood water stool,antifeedant water,body mass decline,extensive necrotic,and exfoliation of intestinal mucosa,chorionic atrophy,short alopecia and multiple hemorrhagic foci with the microscope,all rats of the group died within 9 days.Dose refinement screening:select three dose groups of 11Gy,12Gy,13Gy and the control group.The results showed that the symptoms and signs of the group 13 Gy rats were heavy and the mortality was too high.The symptoms,the gastrointestinal pathology and the survival rate in the 11 Gy group were not significantly different from those in the 12Gy group,however,the time of death distribution in the 11 Gy group was delayed,which was more conducive to the observation and experimental study of traditional Chinese medicine.Finally,the dose of radiation was 11 Gy.2.Pathogenesis and syndrome analysis:in the initial post-irradiation period((about 0?2d),the model group showed signs of restless,polypodiatous and eye red,and the small intestinal mucosa was mild hyperemia and edema.In the early stage(2?7d),the model group showed signs of decrease of body weight,watery stool with blood,intestinal mucosal epithelial multiple hemorrhage and necrotic erosion.In the middle stage(7?13d),after a brief recovery,the quality of rats continued to decline,and unwilling to eat,with massive death.Purple-black mucus was deposited in the gastrointestinal cavity,and submucosal microvascular was occluded.In the early stages of recovery(aboutl 3d?),the rats were cowered with less activity,dry hair,pale paws,and increased interstitial fibers in the intestine.The control group had no abnormalities in living body,gastrointestinal and mucosal tissues.3.Evaluation of clearing away heat,detoxicating and cooling blood method to prevent acute gastrointestinal radiation injury:the mental state,activity,body mass,stool and drinking water of rats in the simple irradiation group all had the characteristic changes of pathogenic heat pathogenic factors,and the number of natural deaths within two weeks was 34,and the traditional Chinese medicine prevention group also had similar characteristic changes,but the symptoms were lighter and the number of natural death was 28.The two groups of animals all showed two period of concentration of death,that is 4?6d and 9?14d.From the condition of survival,the survival rate of the prevention group of traditional Chinese medicine before exposure to 11d was higher than that in the simple irradiation group,and the two groups were close to the trend.4.The mechanism of clearing away heat,detoxicating and cooling blood method for prevention of acute gastrointestinal radiation injury:Compared with the blank control group,the value of NF-kappa BmRNA increased,decreased,increased and decreased in two peak changes,and the value of TNF-alpha showed a gradual decline;the IL-6 value decreased first,then increased again,and the LI-1 beta value increased,descended,then increased,and decreased in two peak changes.The trend of Chinese medicine group was the same as the irradiation group,but at most time,the NF-kappa BmRNA value was lower than the irradiation group,and the lowest at 7d(P<0.01),the decreasing trend of ITNF-? value was relatively slow,and decreased at 1d(P<0.01),increased at 4d(P<0.01),the IL-6 value was lower than that in the irradiated group at each time point,and the lowest in 14d(P<0.01),in addition,the value of 1d was higher than that of the irradiation group(P<0.05),and the value of IL-1 beta in 1d,4d,14d was higher than that of the irradiated group,and the value of 4d and 7d was lower than the irradiated group,and the value at 14d was higher than that of the irradiated group(P<0.05).Compared with the control group,the detection of TXB2 and 6-Keto-PGF1 alpha in the irradiation group showed that the TXB2 value increased,decreased,and then slightly increased.The value of 6-Keto-PGF1 alpha decreased and then decreased after a brief recovery.The value of TXB2/6-Keto-PGF1 alpha decreased slightly,and then gradually increased;The trend of TXB2 and 6-Keto-PGF1 alpha in Chinese medicine group was similar to that in irradiation group.,but Chinese medicine could increase TXB2 at the early stage(0?4d),reduce TXB2 and the ratio of TXB2 to 6-Keto-PGF1 alpha(P<0.05)at the later stage(10?14d).The detection of thromboelastin map showed that the R value in the irradiation group was gradually extended,the K value was shortened,extended and slightly shortened,and the MA value increased,decreased and then slightly increased in the reverse wave type.Compared with the irradiation group,the thrombus elasto graph in the Chinese medicine group was in the same trend,but the R value was shortened at each time point.shorter at 4d,14d(P<0.05),and shorter than the control group(P<0.05)at 1d;MA value increased at 7d,10d and 14d(P<0.05),and increased than the control group at 4d(P<0.05).Conclusions:1.Research on modeling method:using 60Co-gamma as a radioactive source,when the source distance is 3M,the dose rate is 90.65cGy/min-1 and the dose is 11 Gy,the suitable model of TCM can be successfully prepared for the acute gastrointestinal radiation injury.The model can show the characteristic changes of the traditional Chinese medicine inspection,and can be used in the Research of the pathogenesis,syndrome regularity and prevention method of acute gastrointestinal radiation injury.2.Pathogenesis and syndrome analysis:Acute radiation damage of gastrointestinal tract is internal exuberance of heat toxin,blazing of both Qi and blood syndrome in the initial stage;heat burning flesh and blood,Yuan Qi consumption and damage,and devitalization of spleen Yang syndrome in the early stage;Yuan Qi deficiency and damage,true Yin consumption and damage,binding of blood stasis and toxin syndrome in the middle stage;and the liver and kidney depletion and consumption,Qi and blood deficiency,blood stasis and collateral obstruction syndrome in the late stage.3.Evaluation of clearing away heat,detoxicating and cooling blood method to prevent acute gastrointestinal radiation injury:the Chinese medicine can reduce the degree of acute gastrointestinal radiation injury,promote the recovery of the disease,improve the survival state and improve the short-term survival rate,but the preventive medication is not enough to control the development of the disease.4.The mechanism of clearing away heat,detoxicating and cooling blood method for prevention of acute gastrointestinal radiation injury:the Chinese medicine can reduce the expression of inflammatory factors such as NF-kappa B and TNF-alpha in rats with acute gastrointestinal radiation injury in the early stage of irradiation,and reduce the inflammatory reaction of gastrointestinal tract.In addition,it can also effectively improve the coagulation function caused by acute radiation injury,inhibit the microvascular thrombosis and improve the microcirculation.