| Objective:To observe the morphological changes of organs and tissues and the main target organs of poisoning in rats during the poisoning of Paris polyphylla(PPC),and explore the expression of HIF-la and HO-1 in the main target organs of poisoned ratsMethod:Acute toxicity experiments were performed with 56 healthy SD rats,male and female,randomly divided into normal group(n=8),blank control group(n=8)and experimental group(n=40).Distilled water,blank solvent and Different concentrations of ethanol extracts of Paris polyphylla were treated by gavage Observe the general signs of the rats once,record the death,and calculate the half lethal dose(LD50).The rats were sacrificed and the tissues and organs were taken for pathological examination.The morphological changes of the tissues were observed by hematoxylin-eosin staining.For the subacute toxicity experiment,50 healthy SD rats,half male and half female,were randomly divided into the normal group(n=10),the blank control group(n=10),and the experimental group(n=30).General signs of the rats,the rats were sacrificed,the system was dissected,the main poisoning target organs were taken,the histopathological changes were observed by hematoxylin-eosin staining,and HIF-la and HO-1 were detected by immunohistochemistry and Western-Blot.Finally,the statistical analysis method was used to calculate the statistical significance of the differences,so as to understand the effect of the Paris polyphylla poisoning on the levels of various indicatorsResults:1.As a result of the acute toxicity test,the LD50 of the median lethal dose was 43.4 g/kg,and its 95%confidence interval was(38.02,48.98)g/kg.2.The results of HE staining in acutely poisoned rats showed:myocardial tissue agglutination,interstitial congestion,inflammatory infiltration of some myocardial cells,slight hemorrhage;hepatocyte edema,volume increase,hepatic sinus dilatation and congestion,and some hepatocytes with mild fat Degeneration;congestion of interstitial blood vessels in the brain tissue,markedly widened gaps between blood vessels and nerve tissues,glial cell proliferation,slightly widened neuronal fissures,and brain tissue looseness typical of hypoxic lesions;local alveolar expansion,alveolar septal rupture and fusion into Large alveolar cavity,alveolar capillary dilation,congestion,partial alveolar collapse,showing symptoms of hypoxia poisoning;renal tubular congestion is obvious,renal tubular epithelial cells are edema,cytoplasm is lightly stained,and the nucleus is swollen.3.The results of HE staining in subacute poisoning rats showed that the brain neurons of the normal group and the blank control group were neatly arranged,the size was uniform and the distribution was uniform,and there were no obvious abnormalities;the PPC group had loose and edema,interstitial vascular congestion,and vascular and Nerve tissue gaps widened,glial cells proliferated,and neuronal fissures widened slightly,showing typical hypoxic lesions.There was no obvious abnormality of myocardial structure in the normal and blank control groups The muscle bundles in the PPC group were regular,the muscle bundles were slightly wavy,the myocardial lines were blurred and unclear,cytoplasmic agglutination,erythrocyte staining,and some myocardium.The nucleus is lightly stained and fragmented,the myocardial interstitial is slightly widened,and the small interstitial blood vessels are slightly dilated and congested and congested.4.The results of immunohistochemical staining of subacute poisoning rats showed that:?The positive expression of HIF-1? in the brain tissue was mainly localized in the nucleus Compared with the control group,the positive expression of HIF-1? in the horse chestnut and poisonous flower poisoning group was significantly increased Compared with the control group,the positive expression of the low-dose group was significantly enhanced(P<0.05).With the increase of the dose concentration,the expression of HIF-1? decreased,and the positive expression of the high-dose group was significantly reduced,compared with the low-dose group(P<0.01).In the brain tissue,the expression of HO-1 was the strongest in the control group,and the brown particles were significantly more than in the PPC poisoning group.With the increase of the concentration in the poisoning group,the positive expression of HO-1 showed a continuous downward trend,high.The positive signal in the dose group was the weakest,with only a small amount of expression,compared with the control group(P<0.05).?The positive expression of HIF-1? in myocardial tissue was mainly localized in the cytoplasm,the positive expression in the control group was weak,and the brown particles of the positive cells in the poisoning group increased significantly.With the increase of the poisoning dose,the positive expression in the medium dose group and the high dose group was high.In the control group(P<0.05),the positive expression of the high-dose group was significantly enhanced compared with the low-dose group(P<0.05).Compared with the control group,the positive expression of HO-1 in myocardial tissue increased slightly in the poisoning group,the positive expression of HO-1 in the middle-dose group was relatively obvious(P<0.01),and the positive expression in the high-dose group was reduced(P<0.05).5.Western-blot test results of HIF-1? and HO-1 protein expression:? The expression of HIF-la gene in brain tissue showed that the expression in the control group was weak,and the expression in the PPC poisoning group was stronger than that in the control group,of which the low-dose group The expression levels of the high-and medium-dose groups were significantly higher than those of the control group.Compared with the control group(P<0.05),the low-dose group protein expression was down-regulated,slightly higher than that of the control group,and there was a statistical difference compared with the low-dose group significance(P<0.05).With the increase of drug concentration in the poisoning group,the expression of HIF-1? protein decreased.The expression of HO-1 gene in the brain tissue showed that the expression in the control group was stronger,and the expression in the poisoning group was lower than that in the control group.The protein expression in the high-dose group was significantly lower than that in the control group(P<0.05).With the increase of drug concentration,the expression of HO-1 protein gradually decreased.?The expression of HIF-1?gene in myocardial tissue showed that compared with the control group,the low-dose histone expression in the poisoned group was down-regulated,and with the increase of drug concentration,the protein expression was significantly higher than the control group(P<0.05),The expression in the high-dose group was the strongest,and the expression of HIF-1? protein in the poisoned group increased with the increase of the concentration of the drug.The expression of HO-1 gene in myocardial tissue showed that compared with the control group,the expression of the low-dose poisoning group was up-regulated.With the increase of drug concentration,the expression of HO-1 protein increased first and then decreased.The expression level reached the highest when the dose was administered,and the difference was statistically significant compared with the control group(P<0.05).With the increase of the dose,the protein expression decreased significantly,and the protein expression was lower than that of the control group at high dosesConclusions:Rats with Paris polyphylla are poisoned.The median lethal dose LD50 is 43.4g/kg,and its 95%confidence interval is(38.02,48.98)g/kg.The main target organs of poisoning are brain and heart;compared with the control group,the expression of HIF-1? in rat brain and myocardial tissue was consistently up-regulated,but caused The concentration of the drug at the highest expression level was different,indicating that the tolerated doses of organs to PPC poisoning were different Compared with the control group,the expression of HO-1 in the poisoning group is different in brain and myocardial tissues.The expression levels of HIF-1? and HO-1 in myocardial tissues are increased,suggesting that hypoxia induced by PPC can stimulate The expression of HIF-1? and HO-1 protein in myocardial tissue was significantly up-regulated.The high expression of HIF-1? may regulate the expression of HO-1 to reduce the damage of myocardial tissue caused by poisoning and hypoxia. |
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