| Objectives:In this study,the effects of nano liposomes on renal cells of New Zealand rabbits after contrast injection were examined,and the effects of nano liposomes on the prevention and treatment of contrast induced nephropathy(CIN)and the molecular pharmacology mechanism were explained.Finally,it provides important theoretical basis and guidance for the prevention and treatment of CIN.In order to reduce the impact of CIN on the prognosis of patients,reduce the length of stay and hospitalization costs,and provide theoretical basis for the development of new contrast agents for CIN in clinical.Methods:1.Animal experiment I-Establishment and detection of CIN model: establish CIN model,detect the serum creatinine(SCR)and urea nitrogen(BUN)before and 2,8 and 24 hours after the injection of contrast agent iopromide;then kill New Zealand rabbits,and observe the degree of renal injury under electron microscope and light microscope respectively,and comprehensively analyze the changes of SCR and BUN levels before,after the injection of contrast agent No CIN and pathological changes of kidney.2.Animal experiment II-the effect and mechanism of nanoliposomes on the prevention and treatment of CIN: high pressure microjet was used to prepare high-purity linoleic acid nanoliposomes;a New Zealand rabbit model of CIN was constructed,and then it was divided into saline control group,CIN group,contrast +saline prevention group,contrast + nanoliposome prevention group.SCR,bun,Bcl-2,pro apoptotic gene and pro apoptotic gene were detected before,2,8 and 24 hours after the experiment Apoptosis gene(Bax),superoxide dismutase(SOD),neutrophil gelatinase related lipoprotein(NGAL),malondialdehyde(MDA),tumor necrosis factor-α(TNF-α)and other renal functions,as well as the corresponding inflammatory,oxidative stress and apoptosis indexes;to complete the determination of Bax,C3,Fox and p53 in the renal tissue of PCR m RNA;and to observe the renal pathology of New Zealand rabbits under electron microscope and light microscope respectively Thedegree of renal injury was observed.Results:1.Animal experiment I-Establishment and detection of CIN model:(1)The SCR value of the control group was higher than that of the saline control group at 2h,8h and 24 h after the contrast injection,the difference was statistically significant.The bun value of the contrast group was higher than that of the saline control group 24 hours after the contrast injection,the difference was statistically significant.(2)Under the light microscope,the structure of renal cells in the contrast group was vacuolated,the structure of renal tubules was disordered,a few of them were occluded,and a large number of inflammatory cells were seen around them;the rest of them were mitosis and pyknosis;the structure of kidney in the saline group was basically normal.The results of electron microscopy showed that the endoplasmic reticulum and mitochondria of renal unit in CIN group were obviously swollen,the structure of renal lumen was destroyed,and a large number of microvilli were scattered,while the renal structure of saline group was basically normal.2.Animal experiment II-the effect and mechanism of nanoliposomes on the prevention and treatment of CIN:(1)In the New Zealand rabbit animal experiment,in the comparison of the four groups of contrast agent group,saline group,contrast + saline group and contrast +liposome group,the SCR value was significantly different in the comparison of the four groups 8 hours and 24 hours after the contrast agent injection,with statistical significance;the bun value was significantly different in the comparison of the four groups 24 hours after the contrast agent injection,with statistical significance.The incidence of CIN was 0 / 12 in saline group,8 / 12 in contrast group,6 / 12 in contrast+ saline group and 3 / 12 in contrast + liposome group.(2)In the comparison of inflammation,oxidation and apoptosis in ELISA test,there were statistically significant differences in Bcl-2,Bax,NGAL and TNF α indexes among the four groups 24 hours after the contrast injection,and the corresponding indexes of the contrast + liposome group were improved compared with the simple contrast group.The other indicators only have corresponding trend,but there is nostatistical significance.(3)The results of Bax and C3 in the detection of PCR m RNA in renal tissue of the four groups were statistically different,and the corresponding indexes of the contrast agent + liposome group were improved compared with the simple contrast agent group;the rest of the detection results of Fox and p53 had only the corresponding trend,no significant statistical significance.(4)The pathological results were the same as before.The results of electron microscopy showed that the results of CIN group were the same as before.In the corresponding saline + contrast group and liposome+ contrast group,the degree of renal tubules and other pathological structure damage was significantly alleviated compared with CIN group.Conclusion:(1)In this study,we successfully established the CIN model of New Zealand rabbits by injecting iopromide contrast agent into the ear vein of New Zealand rabbits to carry out the experiment,so as to prepare for the further study of the possible pathogenesis of CIN and the role of drug intervention,which plays an important foundation guarantee.Moreover,the results of molding are accurate,stable and highly repeatable.The renal injury of New Zealand rabbits occurred earlier,which may be related to the sensitive response of New Zealand rabbits to the use of contrast medium;however,the creatinine and urea nitrogen levels decreased after 24 hours,indicating that the renal function of New Zealand rabbits began to recover gradually,and gradually recovered to the basic level before operation with the passage of time.(2)By establishing the successful model of CIN New Zealand rabbits,the biochemical and ELISA indexes,the corresponding indexes of PCR m RNA and the pathological changes under the light microscope and electron microscope were compared in four groups: contrast group,saline group,saline + contrast group and liposome + contrast group.Conclusion the first step is to establish a New Zealand rabbit model of CIN successfully,which shows that both liposome and salt water hydration can reduce renal toxicity.According to the above analysis of ELISA and PCR m RNA,we found that the occurrence of CIN was related to inflammatory response,apoptosis and direct renal toxicity.At the same time,it can also show thatnano liposome can prevent and reduce the incidence of CIN by reducing the cell apoptosis damage and inflammatory reaction,and reduce the renal damage caused by contrast agent,which has the same effect as the hydration of salt water.At the same time,the pathological results showed that the renal tubules,endoplasmic reticulum and endothelial cells in the CIN group were severely damaged,and the renal tubules and other pathological structures in the saline + contrast group and the liposome +contrast group were significantly relieved compared with the CIN group.(3)We have done a lot of basic experimental work in this study,but limited to research funding and research time,the sample size of this study is small,which needs to be verified by multi-center and most amount of research work,and also needs to conduct more rich content of basic experimental projects.In addition,the results of the study come from the animal experiments of New Zealand rabbits,which may be quite different from the complex situation of CIN in the actual human body. |
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