| Backgroud:With the development of medical equipment and technology," minimal invasive" became the mainstream in surgery. Endoscopy was widely used in urology because of the minimal invasive, especially for the upper urinary stone, PCNL and URS have gradually replaced the traditional open surgery as minimal scar, shorter hospitalization, better effect of stone remove, faster postoperative recover. High fluid irrigation pressure was needed during PCNL and URS to keep a clear operation field and flush out stone fragments. However, most patients with kidney stones also present with some degree of kidney hydronephrosis, in which, renal parenchyma was thinner then normal ones and have presented a certain of pathology changes. When irrigation pressure was performed during PCNL and URS, whether the irrigation pressure could damage the hydronephrosis kidneys, what level of irrigation pressure could induce damage, whether the toleration is the same for different degrees of hydronphrosis have not been reported at home and aboard. Therefore, we created mild and severe rabbit hydronephrosis model, and fluid irrigation pressure was performed in the model, thepurpose:1. To determined a method of creating mild and severe hdydronephrosis.2. To study what level of irrigation pressure could induced injuries in mild and severe hydronephrosis.3. To study the mitochondrial injury mechanism of mild and severe hydronephrosis underwent various irrigation pressure.Methods:1. A total25New Zealand rabbits were randomly divided into normal group (group N, n=5), mild hydronephrosis group (group M, n=10) and severe hydronephrosis group (group S, n=10). For group N, the left ureter was exposed and dissociated after anesthesia along left lower abdominal cut, then the cut was closed. In group M and S, after anesthesia the left ureter was exposed and dissociated along left lower abdominal cut, a plastic cannula was covered outer of the ureter, then then the cut was closed. All rabbit was respectively examinated by B-ultrasonic after3days for group N and M and after7days for group S. Then rabbits were sacrificed and left kidneys were harvest for Masson dyeing.2. Another54New Zealand rabbits were divided into3groups (Ctrl, M, S) after successfully made the model. For group N, all rabbits irrigation with100mmHg fluid. Both group M and group S were divided into4subgroups (M0-M3and S0-S3), group0to group3were respectively irrigation with0mmHg,20mmHg,60mmHg,100mmHg fluid. After1hour irrigation, the kidneys acute injuries assessed by HE dyeing, TUNEL detection, kidney injury molecule-1, and cysteine-rich61expression using mmunohistochemistry.3. Some part of kidney tissue mitochondrial membrane potential (MMP) were dectected by MMP kits, mitochondrial ultrastructure were observed by electron microscope. The protein were extracted from another part of tissue, Cyt-C, AIF, Caspase-3, Caspase-9, Bcl expression were observed by Western blot.Results:1. In the group N, M and S the paraenchyma thickness were respectively0.36±0.06cm,0.31±0.05cm,0.22±0.04cm. The collection system separation in N, M and S were respectively0.07±0.02cm,0.95±0.17cm,1.70±0.14cm. Both paraenchyma thickness and collection system separation were significant differences among N, M and S (p<0.05). The ratio of fibrosis dyeing by Masson method in N, M and S were respectively0.5%±0.02%,15.4%±0.5%,27.6%±0.7%, and they were significant differences (p<0.05).2. No abnormalities were observed for the Ctrl group, subgroups0, or1(p>0.05). In subgroup2, abnormalities were observed only in the S group, whereas all kidneys in subgroup3suffered acute kidneys injuries, along with occurrence of tubular cells necrosis, increased apoptosis, and increased expression of kidney injury molecule-1and Cyr61(p<0.05).3. No abnormalities of MMP, ratio of mitochondrial vacuolization and changes of apoptosis protein were observed for the Ctrl group, subgroups0, or1(p>0.05). In subgroup2, abnormalities were observed only in the S group, whereas all kidneys in subgroup3suffered acute kidneys injuries, along with occurrence of decreased MMP, increased apoptosis mitochondrial vacuolization, increased expression of Cyt-C, AIF, Caspase-3, Caspase-9, and decreased expression of Bcl (p<0.05).Conclusion:1. It is a valuable method to make mild and severe rabbit hydronephrosis model through plastic cannula obstruction.2. It could induced significant injuries in mild and severe hydronephrosis when irrigation pressure respectively reached100mmHg and60mmHg. High irrigation pressure should be avoided during URS and PCNL.3. Mitochondrial injury mechanism plays a important role in irrigation pressure induce acute hydronephrosis injuries. |
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