The Effects Of Low-energy Laser Irradiation On Histomorphology Of Organs In Pre-eclampsia Of Rat

Pre-eclampsia and eclampsia in pregnancy are unique to the disease,occurred after 20 weeks of pregnancy,manifested by hypertension,proteinuria,edema as the main characteristics of the multiple organ dysfunction syndrome.It is a serious threat to the safety and health of the mother and child,and it is one of the important reasons of essential obstetric death,but its etiology and pathogenesis haven't yet entirely clear. Now it is widely recognized that idiopathic small artery spasm,vascular endothelial injury in pre-eclampsia and eclampsia are the pathogenesis of the central link and the basic feature.Vascular endothelial cells have important endocrine functions through autocrine release of vasoactive factors like nitric oxide,endothelin,prostaglandins to regulate vasomotor regulation,coordinate between coagulation and anticoagulation balance,participate in organizations and the exchange of substances between the blood.When vascular endothelial cell is effected by cytotoxic substances and inflammatory mediators,such as tumor necrosis factor alpha(TNF-α),it can cause vascular endothelial injure,and cause vasoconstriction factor imbalance relaxing factor,in turn there pre-eclampsia and eclampsia is a series of pathological changeto pre-eclampsia and elampsia.Faas,etal first found that low-dose endotoxin(1.0μg/kg) can gradually increase blood pressure of pregnant rats and maintain the high blood pressure,there is a downward trend to the level of post-natal blood pressure,the use of endotoxin can be better prepared animal model of preeclampsia.Experiments have reported that endotoxin could stimulate monocytes, macrophages release TNF-α,TNF-αcan stimulate vasoconstriction substances endothelin release.In recent years,a growing body of research also show that endotoxin stimulation of natural immune system arising from the excessive inflammatory response at the occurrence of preeclampsia,development and transfer process plays an important role.In view of reports of these studies,we select the application of the experiment at low-dose injection of endotoxin in the establishment of the rat model of preeclampsia.Because of the etiology and pathogenisis of pre-eclampsia and eclammpsia are not conclusive,although some progress in the prevention and treatment of pre-eclampsia and eclammpsia has been made,such as the rational use of magnesium sulfate to prevent the poisoning and the occurrence of side effects and anticoagulant protein S,protein C to prevent the application of the dissseminated intravascular coagulation(DIC) the occurrence,but all of the above methods are therapies with Chemical substances and have their side effects,there is no effective new treatment still.In recent years,with low-energy laser on the biological role of the in-depth study.low-energy laser irrdiation has reduced blood viscosity,blood fat,improve red blood cell oxygen carrying capacity,improve the microcirculation and has activase,improve immune function and the role of antixidant function,it has been used in more than 200 kinds of diseases,such as the treatment of clinical cerebral infarction, hypertension,hyperlipidemia,coroary heart disease,diabetes,lung diseases,migraine, hepatitis,rheumatoid arthritis,allergic rhinitis,,and efficacy has been a reliable,at the same time,it is physical therapy and no side effects.In view of the above low-energy laser,and we also have to consider pre-eclampsia and elevated blood pressure changes,we wonder if the low-energy laser will be used in pre-eclampsia- eclampsia is equally effective in the treatment,at present there is no literature at home and abroad Reportedly,this is the purpose of experimental studies.【OBTECTIVE】1,In order to better study the causes of preeclampsia and pathogenesis and new methods of prevention and treatment,the need for stability in an animal model.Faas reports from the research can be established endotoxin rat model of preeclampsia has been a growing body of research tend to the application of endotoxin to establish animal model of preeclampsia.Literature,we compared the advantages and disadvantages of various methods,the choice of the application of low-dose endotoxin rat model of the establishment of pre-eclampsia,for the next phase of research lay the foundation for treatment.2,In view of low-energy laser microcirculation,improve immune function and antioxidant capacity,such as role in the success of model based on the observation of low-energy laser irradiation on rat model of pre-eclampsia and the treatment of pathological changes in organs.【METHODS】1,experimental animalsIn the time of establishment of model,40 adult female SD rats,weighing 200-250 grams,20 adult male SD rats,weighing 250-300 gram.Randomly select 20 female rats with 20 male rats were caged 1:1,the next morning from the sperm found in vaginal discharge to check for pregnancy day 0,and the remaining 20 pregnant female rats no treatment for the control group.In the time of laser irradiation,40 SD pregnant rats,pregnancy and rearing conditions as the same as modeling.2,animal division and model buildingall of rats were divided into 4 groups,non-pregnant group injected with normal saline(NS),pragnancy group injected with normal saline(PS),non-pragnant endotoxin injection group(NE) and pregnancy endotoxin injection group(PE),10 rats in every group.PS group and PE group at 14th day of pregnancy,respectively,along the rat tail vein injection of saline and endotoxin(1.0μg/kg by injection,the concentration of 0.1μg/kg).NS group and NE group on the same day to do the corresponding treatment.3,standard with pre-eclampsia of rat modelAccess to the current reports,blood pressure,urinary protein,offsprings of rat and placenta usually be selected to determine the success of the model,but the specific numerical targets have not yet recognized the unification of standards,since the choice of experimental animal species and different measuring instruments, indicators the basis of value is also very different,by setting the control group in order to compare differences in inter-groups,if there was significant difference among groups then can be defined to the model of pre-eclampsia4,treatment with low-energy laser irradiationAfter the success of building pre-eclampsia model,we use laser irradiate rats and to observe its effect.40 pregnant rats were divided into four groups:saline group, saline-irradiated group,endotoxin group and study group,endotoxin group and study group at 14th day of pregnancy with endotoxin injection as modeling part,saline group and saline-irradiatede group were injected with normal saline,and study group and saline-irradiated group at 16th day began to accept laser irradiation,once a day, every time with 30min,until 19th day of pregnancy.5,monitoring target13th,15th,17th,19th day of pregnancy rat tail artery systolic blood pressure measurement,11th,15th,19th day of pregnancy to collected 24h urine protein content measurement,7th,20th day pregnancy test tail vein blood serum alanine aminotransferase(ALT),blood urea nitrogen(BUN),creatinine(Cr) and TNF-α.At the 20th day of pregnancy,we killed pregnancy rats and taked liver,kidney,lung,heart, brain and placenta,at the same time counting the number and weight of rats and placenta wet weight.6,satistical analysisAll datas were analysed by SPSS 13.0,(?)±S meaned measurement data,the comparison group at different time points using ANOVA for repeated measurement, One-way for ANOVA was used to analyse the information analysis of variance, mutiple comparison tests to meet the homogeneity of variance Bonferroni,method used,does not meet homogeneity of variance test method used Dunnett's T3,to P≦ 0.05(bilateral) have statistically significant difference.【RESULTS】1 results of modeling1.1 systolic blood pressure:the 13th day of pregnancy four group basic blood pressure in rats there were no significant difference(P=0.179),four groups of animals there were comparability;15th,17th,19th day PE increased systolic blood pressure, compared with NS,PS,NE the difference was significant(P＜0.05);NS,PS,NE between the multiple comparisons were not significantly different(P＞0.05);In PE group blood preesure of 13th,15th,17th,19th day there was a significant differences (P＜0.05);NS,PS,NE group,the blood preesure of 13th,15th,17th,19th day there was no significant difference(P＞0.05).1.2urine protein:11th day of pregnancy urine protein in four groups there were no significant differences(P=0.759);the 15th day of pregnancy urine protein in four groups there were significant difference(P=0.048);the differences of pregnancy of 19th day PE and NS,PS,NE were significant(P=0.011),NS,PS,NEcompared with each other there was no significant difference(P＞0.05).1.3serum ALT:7th day of pregnancy rats among 4 groups,there were no significant difference(P=0.578);20th day of prgnancy PE increased compared with the NS,PS, NE there was a significant difference(P=0.002).1.4serum BUN:7th day of pregnancy rats among 4 groups,there were no significant difference(P=0.097);20th day of prgnancy PE increased compared with the NS,PS, NE there was a significant difference(P=0.002).1.5serum Cr:7th day of pregnancy rats among 4 groups,there were no significant difference(P=0.824);20th day of prgnancy PE increased compared with the NS,PS, NE there was a significant difference(P=0.000).1.6serum TNF-α:7th day of pregnancy rats among 4 groups,there were no significant difference(P=0.938);20th day of prgnancy PE increased compared with the NS,PS, NE there was a significant difference(P=0.000).1.7number of rats,body weight and placental wet weight:The number of rat offspring between the two groups was no significant difference(P=0.936),PE rats was significantly lower than the weight of PS group,the difference was statistically significant(P=0.000),PE placental wet weight was significantly lower than PS,the difference was a significant(P=0.011).2 results of laser irradiation2.1 SBP:the 13th day SBP in Four groups was 90-93mmHg,no significant difference (P=0.479);the 15th day of the four groups the overall difference was significant (P=0.000),study group compared with saline group,saline-irradiated group,had a significant difference(P＜0.05),study group and endotoxin group compared SBP,no significant difference(P=0.341);the 17th day among four groups of the overall difference was significant(P=0.000),study group to the saline group and saline-irradiate group,had a significant difference(P＜0.05),study group and endotoxin group compared to SBP,There was a significant difference(P=0.009);the 19th day among the four groups was significant(P=0.000),study group to saline group and saline-irradiate group,there was no significant difference(P＞0.05),study group and endotoxin group compared to SBP,the difference was significant (P=0.000).2.2 urinary protein:the 13th day of urine protein content of the four groups had no significant difference(P=0.648);the 15th day of pregnancy the overall difference between the four groups was significant(P=0.001),in which the study group and the saline group and saline-irradiate group compared urinary protein there was a significant difference(P＜0.05),compared with endotoxin group was no significant difference(P=0.507);the 19th day of overall difference between the four groups was significant(P=0.000),in which urinary protein endotoxin group increased compared with the study group,the difference was statistically significant(P=0.003),the study group and the saline group and saline-irradiate groups of urinary protein content was no significant difference(P＞0.05).2.3 serum ALT:The 7th day of pregnancy in rats among four groups based on serum ALT was no significant difference(P=1.000);the 20th day of pregnancy the overall difference between the four groups was significant(P=0.000),in which the study group ALT and endotoxin group,there was a significant difference(P=0.000),the study group and the saline group and saline-irradiate group,there was no significant difference(P=1.000).2.4 serum BUN:The 7th day of pregnancy in rats among four groups based on serum BUN was no significant difference(P=0.905);the 20th day of pregnancy the overall difference between the four groups was significant(P=0.000),the study group compared with endotoxin group had a significant difference(P=0.000),but with saline group and saline-irradiate group had no significant difference(P=1.000).2.5 serum Cr:The 7th day of pregnancy rats among four groups based on serum Cr was no significant difference(P=0.957);the 20th day of pregnancy the overall difference between the four groups was significant(P=0.049),the study group compared with endotoxin group had a significant difference(P=0.040),the study group and the saline group and saline-irradiate group,there was no significant difference(P=1.000).2.6 serum TNF-α:The 7th day of pregnancy in rats among four groups based on serum TNF-αwas no significant difference(P=0.988);the 20th day of pregnancy the overall difference between the four groups was significant(P=0.000),in which the study group TNF-αand endotoxin group,there was a significant difference(P=0.000), the study group and the saline group and saline irradiate group,there was no significant difference(P=1.000).2.7 number,weight of rats and placental wet weight:between four groups the number of rats was no significant difference(P=0.996);study group in weight of rats and placental wet weight compared with saline group and saline-irradiate group were no significant difference(P=1.000),the study group with endotoxin weight of rats were significantly different(P=0.000),the study group in the wet weight of placenta compared with endotoxin group was significantly different(P=0.021).2.8 Pregnant rats in each group of renal histological observation:saline group and saline-irradiation of normal kidney structure,we can see a complete renal units,of which the normal glomerular and tubular morphology;endotoxin group renal tubular epithelial cell degeneration and necrosis,cell loss tubular lumen narrowing,renal interstitial edema,the study group after laser irradiation became better than endotoxin group.2.9 Pregnant rats in each group to observe the liver histology:saline group and saline-irradiate group liver cells neatly arranged,the district no hyperemia,no expansion of sinusoidal liver cells of normal,non-congestive necrosis,no inflammatory cell infiltration;endotoxin group liver a bit patchy necrosis organization,structure unclear,the liver cells break down,nuclear dissolution,laser irradiation of the study group only had a few spots of necrosis,changes of liver light than endotoxin group.2.10 placenta histological observation of pregnant rats in each group:saline group and saline-irradiated group with placenta decidual thinning,mainly by the fibrous components.Baseband layer of trophoblast giant cells and a small amount of leaf cells,a network of glycogen-like structure,through the maternal blood from entering the labyrinth zone.Lost bands fit into a network terminal villi,forming labyrinth-like structure,the mesh for the gap in maternal blood,maternal and fetal blood is material to the local exchange.Endotoxin group thickened decidua with swelling, inflammatory cell infiltration;baseband IP trophoblast giant cell proliferation,cell island increased vacuolization,thickening of base-band;labyrinth zone trophoblast Ye mast cells,some edema,blood between the membranes by thick,laser irradiation of the study group to alleviate the above.2.11 lung histological observation of Pregnant rats in each group:saline group and saline-irradiated group normal lung tissue structure,the size of the normal alveolar space,endotoxin group thickened alveolar septa and edema,alveolar space smaller vasodilator alveolar wall,there are red blood cells the alveolar exudate,the study group after the laser irradiation to reduce the above.2.12 myocardial histological observation of pregnant rats in each group:saline group and saline-irradiated group of myocardial cells with the rules,the structure of normal muscle cells of small blood vessels,endotoxin group marked interstitial myocardial edema,vascular muscle cells decreased,but myocardial cells,no edema,necrosis,etc., to change,after laser irradiation to reduce edema in the study group.2.13 brain histology observation of pregnant rats in each group:four groups of pregnant rats brain tissue there is no difference between observed and no obvious edema,hyperemia,hemorrhage,necrosis and other pathological changes.CONCLUSIONS1,Pregnancy medium-term low-dose single tail vein injection of endotoxin can increase blood pressure,urinary protein,biochemical indicators of liver and kidney in pregnant rats,but reduce the weight of the placenta and offsprings,these are as same as to the existing standards to identify pre-eclampsia model,and have similar clinical symptoms of the performance with pre-eclampsia,so the model can be considered to be successful.2,In the process of model building,in addition to the above identified indicators of pre-eclampsia,we also found that the model group increased serum TNF-α,liver, kidney,lung,heart and placenta of the morphological changes,in the next step when the laser irradiation will focus on the observed changes in these indicators.3,Low-energy laser irradiation on precordial rat model of pre-eclampsia have a therapeutic effect,it can anti-endotoxin-induced increase blood pressure,urinary protein,liver and kidney biochemical changes and neonatal growth retardation pregnant rats,at the same time,we didn't find that pregnant rats and their offsprings be injured in saline control group,it is an evidence of low-energy laser irradiation therapy as a physical superiority.4,We also found that low-energy laser irradiation reduced serum TNF-αand liver,kidney,lung,heart and placenta pathomorphology change to improve in the model group.Although it can not be confirmed that the pathological changes of organs is the role of endotoxin or secondary changes of pre-eclampsia, indeed,low-energy laser irradiation is effective in improving the pathological changes in various organs.Therefore,the results of this study have provided animal experiments' support for low-energy laser irradiation used in clinical prevention and treatment of pre-eclampsia.