| Background:With the development of minimally invasive technology, Endoscopic surgery has gradually become routine treatment in surgery, especially in the field of urology, along with the rising incidence of urinary tract stones, stones of handling more minimally invasive. All kinds of urureteroscope, nephroscope and flexible ureteroscope has been widely applications along with the progress of medical equipment. It is gradutely replacing the traditional open surgery because of its small incision, shorter hospitalization time, good effect of gravel, fast postoperative recovery characteristics.The majority of doctors and patients become the preferred mode of treatment, however, due to space limitations of endoscopy surgery, the whole operation process can only be carried out within the urinary tract, so to a certain extent, increased the pressure inside the cavity way, especially in the preoperative kidney seeper caused by calculi inherent kidney injury reason, it induced more increased kidney damage when pressure exceeds a certain limit within the pelvis.And then,it caused kidney ischemic damage and acute renal failure.Renal tubule cells mitochondrial permeability transition pore opening is a key link in the pathogenesis of ischemic renal injury,and its subsequent mitochondria mediated apoptosis is an important factor to cause renal tubular cell injury, CsA is the specificity of MPTP inhibitors, many kidney and Extra-renal organs in animal experiments and part of the small clinical studies have confirmed that it can reduce the viscera of ischemic injury by inhibiting MPTP. First of all,we establish different degree of hydronephrosis model in thid topic, before renal pelvis then we perfused drug intervention treatment before pelvic perfusion, The objective is to investigate renal protective effects of Cyclosporine A in treatment of renal based on rabbit models experiencing different degrees of hydronephrosis.Objective:to establish different levels of kidney hydronephrosis model, we explore renal protective effects of Cyclosporine A in treatment of hydronephrosis experiencing renal pelvis perfusion through drug targeting interventions. And we bserve mitochondrial morphological and functional changes after mitochondrial targeted intervention treatment in kidney which exposure to renal pelvis perfusion.Methods:(1) A total of 24 New Zealand white rabbits randomly divided into 3 groups, respectively, the control group (n=6), mild hydronephrosis group (n=9) and severe hydronephrosis group (n=9).For group N, the left ureter was exposed and dissociated after anesthesia along left lower abdominal cut, then the cut was closed? For group M and S, abdomen midline incision exposure were cut apart and the left ureter after the ear intravenous anesthesia.and then dissociate the upper ureter and psoas major, embed the upper ureter 1/4 and two-thirds in the psoas major fissure, then step by step a 3-0 catgut close the abdominal cavity.We make renal ultrasound probe betweenday line double,to observe the kidney seeper and ureter expansion, we observe the left kidney and ureter water expansion after 2 weeks later by ultrasound;Paraffin specimens HE dyeing observation renal tubular damage;Renal ultrastructure change observed under electron microscope.Kidney pathological changes were examined by HE staining; Acute kidney injuries were assessed by analyzing the kidney microstructure.(2) A total of 48 rabbits were randomly allocated into 2 groups, the rabbits underwent a surgical procedure inducing mild (group M, n=24) and severe(group S, n=24) hydronephrosis. In each group, the rabbits were then randomly allocated into 4 subgroups consisting of 6 rabbits each.12 rabbits were gavaged CsA for one week respectively after the success of the molding,one daily. M groups were perfused with 60 and 100 mmHg fluid, respectively.At the same time,S groups were perfused with 20 and 60 mmHg fluid, respectively.Kidney damage assessment were taken 48 hours after perfusion of renal. Kidney pathological changes were examined by HE staining; NGAL expression by using immunohistochemistry. CytC and caspase3 expression by using Western blot.(3) We take some kidney specimens in the second part of the experiment, and extrace mitochondria, transmission electron microscopy test, to observe mitochondria morphology change situation;to observe the mitochondrial membrane potential changes by the mitochondrial membrane potential detection kit line of mitochondrial membrane potential.Results:(1) We probe left kidney by the ultrasound:the control group (N) of the renal pelvis separation was 0.07±0.03 cm, and group M renal pelvis separation was 0.89 ± 0.32 cm, S group of renal pelvis separation was 1.72 ± 0.35 cm, the comparison (p< 0.05),the difference statistically significant;Kidney visible to the naked eye under two group were different degree of swelling, pyelectasis seeper. HE staining under the observed score results showed that N, M and S group score of N (1.8 ± 0.4) respectively;M (23.1± 4.3);S (37.5 ± 3.8), the comparison results have differences, statistically significant difference (P< 0.05);Renal tubular epithelial cells under electron microscope mitochondria swelling and part of the cavity change, the control group did not see obvious mitochondria swelling, mitochondria swelling ratio of M group is lower than S group.HE staining scores were M1'(12.8±3.3);M2'(13.1±4.4); S1'(18.8±3.6); S2'(21.1±1.9) respectively Compared with the experimental groups M1(24.2±5.5); M2(36.5±4.9);S1(39.2±4.8);S2(56.2±4.1), the difference were significant;The expression of Cyt C and caspase3 were decreased compared with the experimental groups, the difference were significant; The expression of NGAL were M1'(2.67± 0.51);M2'(3.17±0.41);S1'(3.33±0.52);S2'(3.50±0.55) respectively compared with the experimental groups M1(4.51±0.55); M2(5.50±0.54); S1(5.67±0.81); S2(6.66±0.51), the difference were significant.(2)Renal ultrastructure change results:electron microscope show that mitochondria of renal tubular epithelial cells are swelling, part of the cavity change, mitochondrial cristae disappear;Corresponding drug intervention group compared with model group, renal ultrastructure change significantly reduce, mitochondria swelling ratio decrease;MMP test results found that:drug intervention group MMP M1'(1.71± 0.31) respectively;M2'(1.54±0.26);S1'(1.66 ±0.17);S2'(1.53±0.15) and control group M1 (1.42± 0.22);M2 (0.88±0.25);S1 (1.39±0.18);S2 (0.81 ± 0.18), drug intervention group compared with control group, the results are different, the difference were significant.(P< 0.05).Conclusion:(1) the psoas major embedding method can accurate effective make mild and severe hydronephrosis model,it began to appear after about a week of hydronephrosis, the embedding method is an effective and accurate method for building.(2) the CsA can reduce the release of the mitochondria in apoptosis factors, inhibiting apoptotic cascade, inhibiting the apoptosis of renal tubule cells, It plays a protective role in treatment of hydronephrosis experiencing renal pelvis perfusion.the CsA plays a protective role in maintain mitochondrial permeability and integrity, it can maintain the stability of the mitochondrial membrane potential and inhibit the occurrence of apoptosis, reduce the damage of renal tubular epithelial cells. |
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