The Influences Of Exogenous Putrescine On The Function And Apoptosis Of Liver And Kidneys In Rats

BackgroundThe acute wound of large area was great menace to patients as same as the chronic wounds such as diabetic and pressure ulcers which were increasing obviously because of upturn living standards and prolonged life. There were always a lot of necrotic tissue in above these wounds, especially, in major deep burns, serious electricity damage, crush injury of limbs and serious wet ulcers of diabetes. In the past, the problems including shock, infection and nutrition were paid attention intensively, but the damage of necrotic tissue to the body were seldom researched. Whereas there were many suspicious phenomena that the local lesion resulted in systemic damage in clinic, for example, the severe burns could induce SIRS, DIC, MODS, etc. In addition, another clinic cachexia phenomenon including emaciation, anemia, hypoproteinemia, even ascites and so on always appeared in serious diabetic foot with wet necrosis that gave us some clue:that is, when the abundant necrotic tissue were absorbed in bloods and result in organism damage and even lead to death finally.Which kind of sphacelus caused organism damage?Researchers presented a lot of component such as burn toxin, endotoxin, lipoprotein complex and lipid peroxide.But some research testified that above component could not lead to rapid non-changeover organism damage. The true damage factors were confused.As all knew, Putrescine and cadaverine existed ether in putrefaction or normal cell. Putrescine and cadaverine in microscale could promote hyperplasia, differentiation and repair of cell, but excessive putrescine and cadaverine could cause serious organism damage, such as SIRS. We once made rabbit model of intra peritoneal injection of putrescine, cadaverine, disinfected homogenate of necrotic tissue and physiological saline and analyzed the influence to inflammation factors in different groups. The experiment found that inflammation reaction tendency in exogenous putrescine group was more approaching to that of homogenate of necrotic tissue group. Thus, putrescine maybe was one of main damage component.In this study,we made a rat model of peritoneal absorbing and selected exogenous putrescine as main damage component to observe the influences of exogenous putrescine on morphological structural and function changes, and apoptosis in liver and kidneys.Objective1.Observe the influences of two concentration of exogenous putrescine on morphological structural and function changes.2.Observe the influences of two concentration of exogenous putrescine on apoptosis of liver and kidneys.3.Record the abnormal activity of exogenous putrescine to rats and compare the difference between the two groups.Materials and Methods1. Experimental animals and groups Ninety of SD rats, Weight (200±20)g, Provided by medicine experimental animals centre of Guangdong province were assigned to three groups randomly including control group(N group,n=10) and low dose(50μg·g-1) group of exogenous putrescine (P1group, n=40),high dose (25μg·g-1)group of exogenous putrescine(P2group, n=40). Selected24h,48h,72h and96h respectively as detecting phase and separately collected ten rats samples and detected them.2. Experiment methods Ip injecting physiological saline2ml, exogenous putrescine (50μg·g-1)2ml and exogenous putrescine (25μg·g-1)2ml and selected24h,48h,72h and96h as detecting separately collected ten rats blood and tissue samples of liver and kidneys.3. Observation index3.1General case record the activity, taking feed, urine volume and colour, faeces colour and smell, Jaundice, death rate in each detecting phase.3.2Biochemistry index Separately collected ten rars blood2-3.5ml, standing30min, centrifuged30min under2500r/min, absorbed out serum, saved in-76℃refrigerator and test Cr, BUN, ALT, AST of serum later by fully automatic Biochemistry analyzer.3.3Pathological change Made HE pathological section and observed histomorphology change.3.4Detected apoptosis of liver and kidneys cellOperated by TUNEL kit instructions and tested apoptosis rate.4. Statistics Measurement data was showed by mean±standard deviation(x±s) and enumeration data was showed by percentage(%),the statistics software of SPSS13.0was employed.This experiment was2X4*4factorial designs,The lines completely randomized two factors factorial analysis (two-way analysis of variance),to understand the main effects and interaction of the various variable;do one-way analysis of variance(One-Way ANOVA) between time groups and treatment groups. Enumeration data used the Pearson Chi-Square way to test. Statistical significance was assumed at p<0.05.Result1. There were transient acute toxicity symptom in P group rats. Wherein, P1group rats appeared spirit restraining change and P2group rats appeared spirit exciting change.There were significant difference between the two groups (χ2=69.408, p<0.01)2. There were few tissue edema and no cell necrosis of liver and kidneys HE pathological section in each detecting phase.3. Value of Cr, BUN, ALT, AST raised in varying degrees in24h,48h,72h and96h detecting phase by TUNEL method, Value of Cr, BUN, AST were highly significant difference between different concentration exogenous putrescine sub-group (F=4.759, P=0.032;F=10.88, P=0.002;F=12.21, P=0.001,P<0.01). Value of Cr、 BUN、 ALT、 AST were significant difference between different detecting phase (F=14.322, P=0.000; F=15.84, P=0.000; F=3.23, P=0.027; F=8.22, P=0.000) and Value of Cr、BUN、 ALT、 AST were highly significant difference between different concentration exogenous putrescine sub-group in different detecting phase (F=5.314, P=0.003; F=9.441, P=0.000; F=16.582, P=0.000; F=11.811, P=0.000).4. Number of apoptosis of liver and kidneys raised gradually and reached at peak value at96h in P1group. Comparing with N group, apoptosis rate of liver at96h in P1group was significant higher [(15.29±1.41)%vs (3.50±0.30)%, P<0.01] and so did the apoptosis rate of kidneys cell at96h in P1group [(24.78±2.19)%vs (4.47±0.33)%, P<0.01]. On the other hand, number of apoptosis of liver and kidneys raised gradually and reached at peak value at48h and fell back normal lever at96h in P2group. Comparing with N group, apoptosis rate of liver at48h in P2group was significant higher[(13.81±1.66)%vs (3.50±0.30)%, P<0.01], and apoptosis rate of kidneys at48h in P2group was significant higher [(26.27±2.13)%vs (4.47±0.33)%, P<0.01].Conclusion1. The exogenous putrescine at certain concentration could lead to functional damage in liver, kidneys in varying degrees,but there were maybe no obvious morphological change.2. The exogenous putrescine could result in rise of apoptosis rate in liver, kidneys. Apoptosis degree was related to dose and time.3. The opposite tendency of apoptosis rate in different concentration showed that the optimum inducing concentration of exogenous putrescine through Ip was between25μg·g-1-50μg·g-14. The functional change of liver and kidneys was related to high apoptosis rate which result from exogenous putrescine.5. Transient non-deadly toxicity manifestation of exogenous putrescine to rats in our research was related to concentration scope of exogenous putrescine.