| Objective: To establish an acute pulmonary embolism animal model by injecting autogenous thrombus into the external jugular vein of rabbits to investigate the expression pattern of HO-1 and 5-HTT in acute pulmonary embolism in rabbits and to analyze the correlation between the expression of HO-Mechanism and mechanism of pulmonary embolism.Methods: In this experiment,36 healthy rabbits were randomly divided into normal control group(n = 4),PE model group(n = 16)and inducer group(n = 16).PE model group rabbits ear vein blood collection,used to make rabbit pulmonary embolism model of autologous thrombus,rabbit external jugular vein catheter injection of autologous thrombus,rabbits appear to deepen the respiration of embolization signs,the extraction of carotid artery blood Detection of oxygen saturation,when the oxygen saturation less than 90%,as modeling success.The induced group was given intraperitoneal injection of cobalt protoporphyrin(COPP)at a dose of 2.5ml / kg.After 24 hours,the models of pulmonary embolism were established.All experimental rabbits were sacrificed at 0h,3h,8h and 24 h after successful model establishment.The lung tissues were removed rapidly.After the changes of the gross tissue of the lung tissues were observed,the rats were adequately rinsed with saline solution,and some of the lung tissues were excised in RNA protection solution.Placed in 4% formaldehyde solution fixed.The morphological changes of lung tissue were observed by HE staining at different time points after successful modeling.The expression of HO-1,5-HTT at different time afterpulmonary embolism was detected by immunohistochemistry and fluorescent quantitative PCR.To investigate the regularity and mechanism of HO-1 and 5-HTT expression in lung tissue during acute pulmonary embolism.Results:1.Under light microscope,the results of HE staining showed that the histological structure of the lung tissue in the normal control group was clearly discerned and arranged uniformly under the light microscope.No emboli were found in the pulmonary artery.In the pulmonary artery,pulmonary embolism showed embolism in the pulmonary artery at various time points,The structure showed varying degrees of hypoxia.After 3h of embolization,infiltration of inflammatory cells and obvious pulmonary congestion and edema were observed in the alveolar cavity.The damage of lung tissue in the induction group was less than that in the pulmonary embolism group.2.Immunohistochemistry results showed that:The expression of HO-1 protein in the normal control group was almost negative.The expression of HO-1 protein in the PE model group was positive,which was significantly higher than that in the control group(P <0.05).The expression of HO-1protein in PE model showed a gradual increasing trend with the extension of time within 24 hours,and there was no obvious increasing trend between 0h and 3h,the difference was not statistically significant(P> 0.05).The expression of HO-1 in induction group was significantly higher than that in normal control group and PE model group(P <0.05),but there was no obvious trend change between the two groups(P> 0.05).The expression of 5-HTT protein in the normal control group was negative.The expression of 5-HTT in the PE model group was positive at 0h,3h and 8h after modeling,which was higher than that of the normal group(P <0.05)Within a first increase,then reduce the trend.After modeling,there was a peak of 5-HTT expression,which was reduced at 3h and dropped to normal after 24h(P> 0.05).The expression of5-HTT protein in the induction group was lower than that in the model group at 0h,3h and 8h(P <0.05),but there was no significant difference between the groups(P> 0.05).3.Fluorescent quantitative PCR test results show: 2-??Ctt obtained values(relative quantification of sample genes)for statistical analysis,said each sample gene is a multiple of the normal group.The expression of HO-1 in each group,compared with the 2-??Ct value of the normal control group(1.000 ± 0),the 2-??Ct value of PE model group was(2.548 ±0.569)times(P <0.05)in 0h group,(3.188 ± 1.186)times(P <0.05)in 3h group,(5.357± 0.750)times(P <0.05)in 8h group,and(9.549 ± 1.391)times(P <0.05)in 24 h group.Compared with the normal control group,the HO-1 of the induction group were(11.144± 1.750),(12.238 ± 1.196),(13.034 ± 1.409)and(13.416 ± 0.767)times,higher than the normal control group at 0h,3h,8h and 24h(P> 0.05).The expression of 5-HTT in each group,compared with the 2-??Ct value of the normal control group(1.000 ± 0),the 2-??Ct value of PE model group was(8.601 ±0.563)times(P <0.05)in 0h group,(6.742 ± 0.951)times in 3h group,(5.691 ± 1.028)times(P <0.05)in 8h group,and(2.610 ± 0.763)times in 24 h group(P>0.05).Compared with the normal control group,the 5-HTT of the induction group were(2.569 ± 0.754),(2.502 ± 0.976),(2.144 ± 0.690)and(2.531 ± 0.766)times,lower than the model group at 0h,3h,and 8h(P <0.05),but there was no statistical significance compared with PE model 24 h group(P> 0.05).Conclusions: In this experiment,the normal group as a control,the PE model group and the inducer group was established,and each group set 0h,3h,8h,24 h time period observation of the pathological changes in each group of lung tissue with HO-1,5-HTT protein expression changes in response to expression trends.1.The morphological changes of all groups in PE model group were significantly changed compared with the normal control group,while in the HO-1 inducer group,although the morphology was still changed compared with the normal group,at the same time,The morphologicalchanges in the model group were small,indicating that the inducer played a protective role in the process of pulmonary embolism.2.Compared with the normal control group,the expression of HO-1 protein in PE model group showed a gradually increasing trend in 24 h,indicating that the expression of HO-1 increased with pulmonary embolism,and immunohistochemistry The results showed that the brown particles mainly concentrated in pulmonary artery endothelial cells,pulmonary artery smooth muscle cells and alveolar macrophages,indicating the expression of HO-1 and vascular association.Compared with the normal group,the expression of 5-HTT protein showed a trend of first increase and then decrease,indicating that the expression of 5-HTT was inhibited with the extension of time in the process of pulmonary embolism.In the inducer group,5-HTT expression was significantly reduced compared with the normal group,indicating that the expression of 5-HTT in pulmonary embolism may be inhibited by the expression of HO-1.3.The results of immunohistochemical tan positive staining and fluorescence quantitative PCR showed that 5-HTT showed a decreasing trend with the increase of HO-1,the interaction between the two in the occurrence and development of pulmonary embolism The process of great significance,the pathogenesis of pulmonary embolism and disease prevention and treatment has a very important role. |
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