Experimental Study On The Effect Of Shenfu Decoction On Survival Time And Mortality Of Hypothermia Rats Soaked In Seawater

BackgroundHypothermia is a common cause of death of people who fall into the sea in war and trade at sea.So far,a reasonable and effective rewarming method is an important measure to treat hypothermia patients.Common rewarming measures can be divided into passive rewarming or active rewarming,which can help people who fall into the sea to return to normal body state.However,it is difficult to rescue at sea,and the acute and severe hypothermia caused by long-term seawater immersion is accompanied by serious complications after rewarming,which seriously harms human health.At present,even in large hospitals,the number of patients with seawater immersion hypothermia treated every year is still small,so it is impossible to carry out clinical trials.Therefore,the establishment of animal model is a key step to study hypothermia caused by seawater immersion.In recent years,the application of traditional Chinese medicine may bring new hope for the treatment of hypothermia.The etiology and pathogenesis of hypothermia caused by seawater immersion in traditional Chinese medicine are excessive cold pathogen,internal viscera and yang failure,and the treatment should be to restore yang and solidify it.Shenfu decoction has the effects of invigorating qi,restoring yang and strengthening the body.It is not clear whether Shenfu decoction can be used to treat hypothermia caused by seawater immersion,which is worth exploring.Objections1.Establish a platform for hypothermia in rats by seawater immersion,and observe the effect of seawater immersion on vital signs of rats.2.Shenfu decoction was prepared based on the ratio of Red Ginseng to Monkshood aconite tablets =1:1,and the model was established immediately after a single oral administration to explore the influence of Shenfu decoction on the survival time and mortality of hypothermia rats soaked in seawater.MethodsPart I: Establishment of experimental platform for hypothermic rats immersed in seawater.1.Platform construction of hypothermic rats immersed in seawater: including self-made rat vertical fixer,self-made rat seawater immersion fixed water tank,automatic constant temperature controller for heating and cooling and the overall framework of the platform.2.Implantation of rat temperature recorder: Set the temperature monitoring task of DS1922 L electronic temperature recorder,and implant it into male and SPF SD rats to monitor the core temperature of rats.3.After the rats implanted with the temperature recorder were raised for 10 days,the rats with good growth were selected as experimental animals and randomly divided into seawater immersion group and normal control group;Establish a platform for rats with hypothermia by seawater immersion.Rats in seawater immersion group were immersed in 15 ℃ artificial seawater until they died.During the immersion,the physiological changes of rats were observed,the activity consciousness grade of rats was recorded,and the survival time and mortality rate of rats were monitored.The rats in the normal control group were fixed in a fixed water tank without artificial seawater immersion at the same room temperature for 24 hours;After the experiment,the temperature recorders of the two groups of rats were taken out and the data were derived.Samples were taken and stained with HE to evaluate the pathological changes of liver,spleen and other organs in two groups of rats.Part II: Experimental study on the effect of Shenfu decoction on survival time and mortality of hypothermia rats immersed in seawater.1.Building a platform for seawater immersion in rats with hypothermia.2.Configuration of Shenfu Decoction: Search China National Knowledge Infrastructure Database(CNKI),Pub Med and other websites to sort out the composition and clinical application of Shenfu Decoction in modern times;Mix Panax ginseng and Radix Aconiti Lateralis Preparata according to the ratio of 1:1,and decoct in a decocting pan;Prepare Shenfu decoction with different concentrations;The components of Shenfu decoction were analyzed by ultra performance liquid chromatography-high resolution mass spectrometry(UPLC-Q-TOF/MS).3.Effects of different doses of Shenfu decoction on survival time and mortality rate of hypothermia rats immersed in seawater: Male,SPF and SD rats were randomly divided into seawater immersion control group,Shenfu decoction group of 0.675 g/kg,Shenfu decoction group of 1.35 g/kg,Shenfu decoction group of 2.7 g/kg,Shenfu decoction group of 5.4 g/kg and Shenfu decoction group of10.8 g/kg.Rats were given different doses of Shenfu decoction or distilled water once,and then placed in 15℃ artificial seawater to establish a hypothermia model immediately.During the immersion process,the simple classification of rat activity consciousness was recorded,and the survival time and mortality rate of rats were monitored.4.Effect of Shenfu decoction on pathological and hematological indexes of hypothermia rats immersed in seawater: Male,SPF and SD rats were randomly divided into 4 groups,namely 1.5 h control group,1.5 h Shenfu decoction group,3 h control group and 3 h Shenfu decoction group,with 10 rats in each group.Rats were given Shenfu decoction(1.35 g/kg)or distilled water once,and then the hypothermia model was established in 15℃ artificial seawater.Immediately after soaking,blood and important organ samples of rats were collected.The blood routine,liver function,renal function,cardiac function and inflammatory factors in rats were detected.HE staining was used to evaluate the pathological changes of liver,heart,spleen,lung,kidney and other organs in the two groups.ResultsPart I: Establishment of experimental platform for hypothermic rats immersed in seawater.1.With the prolongation of immersion time,the activity,respiration and heart rate of rats in seawater immersion group decreased,and they gradually entered syncope state.2.Before seawater immersion at low temperature,there was no significant difference in core temperature between normal control group and seawater immersion group(P =0.475).In the normal control group,there was no obvious change in the core temperature during fixation.After 10 min,the average core temperature of rats in seawater immersion group was lower than 32 ℃(P < 0.001).After 20 min,it was lower than 28 ℃(P <0.001);After 40 min,it was lower than 25 ℃(P < 0.001);After 70 min,it was lower than 21 ℃(P < 0.001);After 170 min,the average core temperature was close to the water temperature(P < 0.001).3.In the normal control group,there was no obvious change in consciousness grade during fixation,and there was no significant difference between the two groups when immersed in seawater for 30 min(P = 0.317).After rats were immersed in seawater for 1 h,there was a significant difference between the two groups,and the simple classification of activity consciousness of rats in seawater immersion group decreased(P < 0.001).4.Rats in the normal control group survived within 24 h,while rats in the seawater immersion group began to die at 3.77 h,with 50 %death time of 13.53 h and 100 % death time of 16.77 h.5.Compared with normal rat organs,the liver,heart,spleen,lung,kidney,stomach and small intestine of dead rats have significant pathological changes after seawater immersion.Part II: Experimental study on the effect of Shenfu decoction on survival time and mortality of hypothermia rats immersed in seawater.1.After literature retrieval,72 documents were finally included.The literature results show that the proportion of Shenfu decoction is 1:1;Panax ginseng is the only kind of ginseng in Shenfu Decoction.There are many kinds of processed aconite in Shenfu decoction,but the proportion of them is small.We can choose monkshood that can only be obtained in hospitals and pharmacies to prepare the required Shenfu decoction.Shenfu decoction is mostly based on clinical research and less on basic research;Diseases involve multiple systems.2.UPLC-Q-TOF/MS method showed that Shenfu Decoction contained 50 kinds of compounds,such as Rubric acid,polybenzoyl aconitine,ginsenoside Rg1,ginsenoside Rf and ginsenoside Rh1.3.The simple classification of activity consciousness of rats showed that the simple classification of activity consciousness of rats in the 1.35 g/kg Shenfu decoction group increased at 1 h and 2 h compared with the seawater immersion control group,with grades 4 and 5(P <0.001)and grades 2 and 3(P < 0.001)respectively,and there was no significant difference between the simple classification of activity consciousness of rats in the other doses of Shenfu decoction group and the control group.4.Record the survival time of each rat in seawater immersion.Compared with the average survival time of the seawater immersion control group of 10.88 1.98 h,the survival time of the 5.4g/kg Shenfu decoction group was 15.22 0.63 h(P = 0.047)and that of the 1.35g/kg Shenfu decoction group was 16.94 1.42 h(P = 0.006).Further survival analysis showed that compared with the seawater immersion control group,the difference was statistically significant except1.35g/kg Shenfu decoction group(P = 0.015),and the survival rate of rats in other doses of Shenfu decoction group was not significantly improved.5.Compared with the seawater immersion group for 3 h,the mean hemoglobin content(MCH),mean hemoglobin concentration(MCHC),platelet(PLT)decreased significantly(P < 0.001),while the values of total protein(TP),alkaline phosphatase(ALP),glucose(Glu),uric acid(UA)and creatinine kinase(CK)increased significantly(P < 0.001).Compared with the control group,the values of red blood cell count(RBC),hematocrit(HCT),aspartate aminotransferase(AST),creatinine(Cr)and UA of rats in the traditional Chinese medicine group decreased under the conditions of 1.5 h and 3 h seawater immersion(P <0.05),while the values of GLU all decreased.Under the condition of seawater immersion for 1.5 h,compared with the control group,the values of AST,CR,UA,lactate dehydrogenase(LDH)and interleukin-10(IL-10)in the Chinese medicine group were significantly decreased(P < 0.05).Under the condition of 3h seawater immersion,compared with the control group,the values of AST,TP,CR,UA,interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in the Chinese medicine group were significantly decreased(P < 0.05).6.The results of HE staining showed that the stomach,lung,liver,kidney and spleen of rats showed pathological changes in different degrees after seawater immersion.Compared with the control group,there was no obvious difference in the pathological changes of rats in the traditional Chinese medicine group under the condition of 1.5 h immersion,and the degree and range of pathological changes in the stomach were significantly decreased under the condition of 3 h seawater immersion.Under the condition of 1.5 h seawater immersion,the lung bleeding of rats in the Chinese medicine group was improved compared with that in the control group,while under the condition of 3 h seawater immersion,there was no significant difference between the Chinese medicine group and the control group.Conclusion1.The platform of seawater immersion hypothermia in rats was successfully established.The physiological status of rats with seawater immersion hypothermia changed,the core temperature and consciousness grade decreased,the survival time was significantly reduced,and typical pathological injuries appeared.2.1.35 g/kg Shenfu decoction with the ratio of Red Ginseng: Monkshood =1:1 has remarkable curative effect on hypothermia rats soaked in seawater,which can improve the simple classification of activity consciousness,survival time and survival rate of rats,which may be related to improving blood system,liver,kidney and immune system and alleviating pathological damage of stomach and lung.