| ObjectiveTo investigate the effect of different pressure of CO2pneumoperitoneum, time of CO2pneumoperitoneum on renal tissue of rabbits histological, proliferating cell nuclear antigen (PCNA),cell apoptosis,ultrastructural and superoxide dismutase (SOD), malondialdehyde (MDA). To investigate the effects of methylprednisolone sodium succinate on expression of PCNA, SOD activity and the quantity of MDA in renal tissue when the nephrectomy was undergone in different CO2pneumoperiton-eum laparoscopic.Methods65rabbits were randomly divided into control group (5rabbits), No medicine group (30rabbits), Treatment group (30rabbits). The control group(5rabbits) only channel establishment, not to the intraperitoneal injection of CO2pneumoperitoneum; In the No medicine group (30rabbits)and treatment group (30rabbits), according to the experimental conditions the two groups were divided into6groups,5rabbits in each group,namely5mmHg(5rabbits),7mmHg(5rabbits),9mmHg (5rabbits), pneumoperitoneum time set for2hours; Different time of pneumoperitoneum group was1hours (5rabbits),2hours (5rabbits),3hours (5rabbits), pneumoperitoneum pressure for7mmHg. Operation method:3%pentobarbital sodium1ml per kilogram was given by the ear vein and the dosage was adjusted according to the operation time, then the animal was fixed on the animal experiment platform in supine position after anesthetic effecting. anesthetic could be properly added to complete the operation in accordance with the pressure and time of the experimental set. The animals were injected the methylprednisolone sodium succinate on0.5mg/Kg by the ear vein before the establishment of CO2pneumoperitoneum half an hour in the treatment group. Laparoscopic cut the left kidney was done. Light (HE stain) and electron (lead citrate/uranyl acetate stain) microscopic histological change of the renal glomerulus and tubule in different pneumoperitoneum condition was observed. The expression of PCNA immunohistochemical of renal tubular, glomerular was determined in semi-quantitative by medical image management and analysis system. The counts of the cell apoptosis was determined by TUNEL method. The activity of SOD and the quantity of MDA were mearsured by xanthine oxidase and thiobarbituric acid respectively. The change of PCNA, SOD, MDA and cell apoptosis in each groups of different pneumoperitoneum conditions were compared with SPSS12.0software.Results1. The quantity of expression of PCNA of the glomerular, tubular in control group was8.13±1.98,9.14±2.07, respectively; Cell apoptosis was6.13±1.68,7.34±1.17, respectively; SOD activity was375±13.5; The quantity of MDA was8.01±1.60.2. The quantity of expression of PCNA of the glomerular, tubular in5mmHg,7mmHg,9mmHg of the no medicine groups was6.74±2.11,8.32±1.97;5.32±1.94,6.59±2.17;4.96±2.02,5.27±1.66, respectively. Cell apoptosis was26.74±2.11,18.32±1.91;29.32±1.94,23.29±2.77;34.16±2.02,29.27±3.66,respectively. SOD activity was317±11.3,294±9.04,251±10.7,respectively. The quantity of MDA was9.35±1.07,14.7±2.31,18.4±1.93,respectively.The above data of each group were compared with the control group and all the P were<0.05; Comparison within group P<0.05.The quantity of expression of PCNA of the glomerular, tubular in1h,2h,3h of the no medicine groups was7.32±1.90,8.98±2.15;5.32±1.94,6.59±2.17;4.41±1.52,6.38±1.64, respectively. Cell apoptosis was27.32±1.90,19.92±3.17;29.32± 1.94,23.29±2.77;36.41±1.52,26.38±2.64,respectively.SOD activity was331±12.4,294±9.04,265±8.91, respectively. The quantity of MDA was8.97±1.38,14.7±2.31,20.2±1.69, respectively.The above data of each group were compared with the control group and all the P were<0.05; Comparison within group P<0.05.With pneumoperitoneum pressure increasing, time extending in no medicine group, Renal pathologic sections showed glomerular and tubular mild hyperemia, edema, infiltration of inflammatory cells were occasionally observed. Tunel films showed glomerular, tubular apoptosis cells increased. Cell ultrastructural section showed that mitochondria increased and irregular, edema, space narrowed, disintegrated pieces a little part of body, mitochondria vacuolated, lysosomal a little, the nuclear membrane was structured, Chromatin is granular, not evenly distributed in the nucleus.3. The quantity of expression of PCNA of the glomerular, tubular in5mmHg,7mmHg,9mmHg of the treatment groups was9.78±2.03,10.7±1.05;12.8±2.07,13.3±2.57;14.3±2.31,15.1±2.67, respectively. Cell apoptosis was19.78±2.03,20.70±1.15;16.83±2.07,18.33±2.57;15.36±2.41,15.19±2.17,respectively.SOD activity was397±10.2,439±7.81,470±11.9,respectively. The quantity of MDA was8.03±0.94,7.91±1.02,7.13±1.49,respectively.Comparison within group P<0.05. Comparison with the control group and all the P were<0.05;Comparison with the no medicine group and all the P were<0.05.The quantity of expression of PCNA of the glomerular, tubular in1h,2h,3h of the treatment group was10.3±3.11,11.9±1.99;12.8±2.07,13.3±2.57;15.7±1.92,16.3±1.71,respectively.Cell apoptosis was19.34±3.71,18.94±1.19;16.80±2.47,17.31±2.57;17.73±1.52,15.35±1.71,respectively.SOD activity was379±9.33,439±7.81,494±10.7,respectively.The quantity of MDA was7.98±1.21,7.91±1.02,6.85±0.93,respectively.Comparison within group P<0.05. Comparison with the control group and all the P were<0.05.Comparison with the no medicine group and all the P were<0.05.With pneumoperitoneum pressure increasing, time extending in treatment group, renal pathologic sections showed glomerular, tubular mild hyperemia, edema, infiltration of inflammatory cells were occasionally observed. Tunel films showed glomerular, tubular apoptosis cells decreased. Electron microscopic sections showed that mitochondria increased and irregular, gap widened, disintegrated pieces a little, rare mitochondrial vacuolization phenomenon, the nuclear membrane structure, chromatin filaments helical columnar, rod.Conclusions1. With the CO2pneumoperitoneum pressure increasing and time extending, SOD activity reducted, Oxygen free radical generated, released and its metabolites MDA increased; the expression of PCNA reduced and the number of renal cell apoptosis increased.2. The renal ischemia reperfusion injury, Laparoscopic nephrectomy in rabbits as donor kidney, maybe protected by Methylprednisolone sodium succinate in a certain degree by means of making the SOD activity increase, MDA ease, expression of PCNA increase and apoptosis of renal cells reduce after the use of it. |
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