The Effect Of Salvia Miltiorrhiza On Renal Injury Of Hydronephrotic Kidneys In Neonatal Rat With Partial Ureter Obstruction

Objective:To investigate the effect of Salvia Miltiorrhiza injection on renal injury of hydronephrotic kidneys in neonatal rat with partial ureter obstruction, so as to provide reliable experimental and theoretical basis of better clinical treatment for the congenital hydronephrotic kidneys in children.Methods:1、To make animal model120newborn wistar rats in48hours were randomly divided into5groups,24wistar rats each.To randomly selected1group line sham operation,postoperative three week began to intraperitoneal injection of2.0ml·kg-1·d-1physiological saline as sham group, and the other4groups separated into experimental group、low doses of salvia miltiorrhiza group (intervention group1)、dose of salvia miltiorrhiza group (intervention group2) and high doses of salvia miltiorrhiza group (intervention group3). The experimental group is to make the left partial ureter obstruction (PUO) by psoas oppression ureter, postoperative3weeks the model group and the intervention group1,2,3respectively began to intraperitoneal injection of2.0ml·kg-1·d-1physiological saline、the mixture1(salvia miltiorrhiza injection and physiological saline ratio is1:3), the mixture2(salvia miltiorrhiza and physiological saline injection of the ratio of1:1)、salvia miltiorrhiza injection; The animals of every group have random divided into three subgroups, in feeding five weeks, seven weeks and ten weeks each group of animals randomly selected one subgroup to be sacrificed, cut off renal tissues and collect urine samples inspected.2%To examine various kinds maker(1), To examine kidney disease organization of each different period (after established partial obstruction model intervention five weeks, seven weeks and ten weeks) by HE staining, and using transmission electron microscope (tem) observe the ultrastructural changes;(2), To examine the transforming growth factor-β1(TGF-β1) expression of kidney disease organization in different periods through the immunohisto chemical PV method, and application of Image-ProPlus Image analysis software average light density measurement value; To examine TGF-β1mRNA level chage in the kidney tissues by the RT-PCR technology;(3), To examine TGF-β1level in renal pelvis urine of different periods application ELISA test.Results:1、The survival of the animals in animal model establishment period: Only120newborn rats, die7in establish hydronephrosis obstruction model and feed period, live113(94.17%)(anesthesia death only1, intraoperative death only4, feeding death2).2、The changes in organization morphology of the kidney disease:(1)、The change of the organization structure of kidney disease by the naked eye view and light microscopy:by the naked eye view of the control group in different feeding period,has bilateral renal size, shape is consistent, bilateral renal glomerulus, renal tubular developing normally, glomerular full, renal tubular has not seen the expansion; The model group with the fed prolonged, kidney disease than health renal increase gradually, expansion of the renal pelvis, water retention, renal glomerulus disease, renal tubular obvious dysplasia, glomerular bowman surname bag expansion, renal tubular expansion. The same intervention group with the feeding time extension, the kidney disease by naked eye view is still more than healthy kidney, under mirror glomerular, renal tubular damage over time is extended increase trend, but pathology classification was significantly lower than the model group (P<0.05); But in the same feeding time, along with the increases salvia miltiorrhiza doses of the intervention group, kidney disease organization structure damage gradually ease.(2)、The ultrastructural of kidney tissues were observed under transmission electron microscope:the control group glomerular epithelial cells form, structure normal, were not see the epithelial cells of the abnormal, renal tubular epithelial cells mitochondria crest clearly visible; Model group see glomerular epithelial cells form, structure apparently unusual, lodging and foot processes are segmental fusion, or even fall off loss, proximal tubule mitochondria swelling, empty bubble of the kidney, small tube can be seen in a large number of cavity cellular debris; Intervention group kidney disease ultrastructural are different degree of change, with feeding the extension of time damage increase gradually, but compare with the model group light, the same feeding time points with the increase of salvia miltiorrhiza dose glomerular epithelial cells form structure damage to reduce.3、The TGF-β1expression in the kidney tissues:The TGF-β1expression in the kidney tissues of the control group is lower, the TGF-β1expression of the model group high expression in renal tubular, TGF-β1mRNA expression is raised. The TGF-β1expression in the kidney tissues of every intervention group are different in degree,and the same intervention group with the feeding time extension TGF-β1expression of the quantity is increasing gradually, but still significantly lower than the model group; Comparison between the TGF-β1expression in the kidney tissues of the same intervention group different feeding time point (feed five weeks, seven weeks, ten weeks),low doses of salvia miltiorrhiza in the group are statistically significant differences (P<0.05), high dose group and dose group of five weeks and seven weeks in comparison, have not statistically significant differences (P>0.05); The TGF-β1expression in the same time after intervention has gradually reduced trend with the increase of salvia miltiorrhiza dose, but higher than the control group. By comparison between the control group, the model group and the intervention group, the differences are statistically significant (P<0.05). Comparison between the same feeding time different doses of salvia miltiorrhiza (low, medium and high dose), establish five weeks in obstruction, the difference was not statistically significant (P>0.05), the more different time points comparative differences between are statistically significant (P<0.05).4、The TGF-β1level changed in the renal pelvis urine of kidney diseaseThe TGF-β1content in the urine of the control group is low. The model group kidney disease is high. The content in every intervention group kidney disease is between in the control group and model groups. By comparison between the TGF-β1level in urine of the control group, the model group and the intervention group, the differences are statistically significant (P<0.05). By comparison between the TGF-β1expression in urine of the same intervention group different feeding time point (feed five weeks, seven weeks, ten weeks), statistical significant is difference in low doses of salvia miltiorrhiza(P<0.05), high dose group and dose group of five weeks、seven weeks in comparison, have no statistical significance differences (P>0.05); The more different time points comparison between groups was statistically significant difference (P<0.05).The same breeding time points, comparison between the TGF-β1level in urine of intervention group establish five weeks obstruction, there were no difference in statistical significance (P>0.05). Compared between the TGF-β1level in urine after obstruction was set up seven weeks, ten weeks, are statistically significant difference (P<0.05).Conclusion:1、Salvia miltiorrhiza injection can prevent or reduce newborn mice renal damage of hydronephrosis disease. It has protective effect.2、Through testing the TGF-β1level in kidney tissues and urine of animal model newborn mice hydronephrosis indicates:the protection of salvia miltiorrhiza injection about newborn rat hydronephrosis disease renal damage has relation between time and effect. To infer salvia miltiorrhiza of clinical application has protection on renal injury of hydronephrotic kidneys in children.