PPARγ配体对脓毒症急性肾损伤保护作用的研究

发布时间：2018-05-03 10:56

本文选题：脓毒症 + 急性肾损伤　；参考：《大连医科大学》2014年硕士论文

【摘要】：目的：探讨罗格列酮（Rosiglitazone，ＲＯＳＩ）——PPARγ（Peroxisomeproliferators-activated receptor γ，过氧化物酶增殖激活受体）的配体在脓毒症急性肾损伤时对大鼠肾脏结构和功能的影响，以及凋亡调控基因Bcl-2在脓毒症肾脏组织中表达水平的变化，为PPARγ在脓毒症急性肾损伤中的保护作用提供实验依据。方法：雄性SD大鼠96只随机分为4组，分别为盲肠结扎穿刺加罗格列酮给药组（ＣＬＰ+ＲＯＳＩ组）、盲肠结扎穿刺组(ＣＬＰ组)、假手术加罗格列酮给药组(Sham+ＲＯＳＩ组)、假手术组(Sham组)。采用盲肠结扎穿刺法（Cecal ligation andpuncture，ＣＬＰ）建立脓毒症模型，ＣＬＰ+ＲＯＳＩ组、ＣＬＰ组术前30分钟分别腹腔注射罗格列酮（ＲＯＳＩ，10mg/kg）或等体积的药物溶剂10%二甲基亚砜（DMSO）； Sham+ＲＯＳＩ组、Sham组术前30分钟分别腹腔注射罗格列酮（ＲＯＳＩ，10mg/kg）或等体积的药物溶剂10%二甲基亚砜（DMSO）。各组分别在术后0，6，12，24小时各取6只大鼠腹主动脉取血，分离血清检测肌酐和肿瘤坏死因子α，观察各组大鼠一般状态并应用HE染色检测肾脏病理组织学变化，检测肾脏组织的髓过氧化物酶活性（MPO），应用Western blots测定PPARγ及Bcl-2凋亡相关蛋白在肾脏组织中的表达。采用SPSS13.0软件包对资料进行t检验中成组设计两样本均数的检验，P0.05为差异具有统计学意义。结果：ＣＬＰ组肌酐水平随时间的延长而逐渐升高，ＣＬＰ组与Sham组之间6h肌酐浓度比较无明显统计学差异（P6=0.812，P0.05），12h、24hＣＬＰ组与Sham之间血清肌酐浓度比较明显升高（P12=0.047，P24=0.001，P0.05）；ＣＬＰ+ＲＯＳＩ组与Sham+ＲＯＳＩ组之间6h肌酐浓度比较无明显变化（P6=0.868，P0.05），12h、24hＣＬＰ+ＲＯＳＩ组与Sham+ＲＯＳＩ组之间血清肌酐浓度比较明显升高（P12=0.033，P24=0.039，P0.05）；0h及6hＣＬＰ+ＲＯＳＩ组与ＣＬＰ组之间血清中肌酐浓度比较无明显变化（P0=0.418，P6=0.528，P0.05），在12h及24hＣＬＰ+ＲＯＳＩ组与ＣＬＰ组血清肌酐浓度比较，ＣＬＰ+ＲＯＳＩ组血清肌酐的升高较ＣＬＰ组明显减少（P12=0.04，P24=0.006，P0.05）。6h、12h、24hＣＬＰ组与Sham之间血清TNFα浓度比较有明显升高（P6=0.00,P12=0.00，P24=0.00，P0.05）；ＣＬＰ+ＲＯＳＩ组与Sham+ＲＯＳＩ组之间6h、12h、24h血清TNFα浓度比较有升高（P6=0.00,P12=0.00，P24=0.00，，P0.05）；6h、12h及24hＣＬＰ+ＲＯＳＩ组与ＣＬＰ组血清TNFα浓度比较，ＣＬＰ+ＲＯＳＩ组血清TNFα浓度升高的水平较ＣＬＰ组明显降低（P6=0.01，P12=0.02，P24=0.01，P0.05）。0h及6hＣＬＰ+ＲＯＳＩ组与ＣＬＰ组之间肾脏组织髓过氧化物酶活性比较无明显统计学差异（P0=0.875，P6=0.13，P0.05），在12h及24hＣＬＰ+ＲＯＳＩ组髓过氧化物酶活性升高的水平较ＣＬＰ组有明显减少（P12=0.01，P24=0.006，P0.05）。ＣＬＰ组髓过氧化物酶活性明显高于Sham组（P6=0.00,P12=0.00，P24=0.00，P0.05）。ＣＬＰ+ＲＯＳＩ组与Sham+ＲＯＳＩ组之间肾脏组织髓过氧化物酶活性比较有升高（P6=0.00,P12=0.00，P24=0.00，P0.05）。与Sham组和Sham+ＲＯＳＩ组相比，ＣＬＰ+ＲＯＳＩ组、ＣＬＰ组均出现大鼠脓毒症症状，以及器官功能障碍，而ＣＬＰ+ＲＯＳＩ组的大鼠症状及器官功能障碍程度较ＣＬＰ组减轻。通过Westernblots测定各组PPAR-γ以及Bcl-2的水平，ＣＬＰ组PPARγ及Bcl-2的表达较ＣＬＰ+ＲＯＳＩ组明显降低。结论：罗格列酮作为配体激活PPARγ，从而减轻了脓毒症时的炎症反应，减少肾脏细胞凋亡，改善了肾脏功能，减轻肾脏损伤，对大鼠脓毒症急性肾损伤有保护作用。
[Abstract]:Objective: To investigate the effect of Rosiglitazone (ROSI) - PPAR gamma (Peroxisomeproliferators-activated receptor gamma, peroxidase activation receptor) on the renal structure and function of rats with acute renal injury, and the expression of Bcl-2 in the renal tissue of sepsis. The changes provide an experimental basis for the protection of PPAR gamma in septic acute kidney injury.
Methods: 96 male SD rats were randomly divided into 4 groups, which were cecum ligation and rosiglitazone group (group CLP+ROSI), cecum ligation group (group CLP), sham operation plus rosiglitazone administration group (group Sham+ROSI), sham operation group (group Sham), Cecal ligation ANDPUNCTURE, CL (CL). P) establish a sepsis model, group CLP+ROSI, group CLP, 30 minutes before operation, intraperitoneal injection of rosiglitazone (ROSI, 10mg/kg) or equal volume of drug solvent 10% two methyl sulfoxide (DMSO); group Sham+ROSI, group Sham (ROSI, 10mg/kg) or equal volume of drugs, respectively, 30 minutes before operation. The solvent 10% two methyl sulfoxide (DMSO). Each group took 6 rats' abdominal aorta at 0,6,12,24 hours after the operation to extract the blood from the abdominal aorta. The serum creatinine and tumor necrosis factor alpha were isolated from the serum. The general state of the rats was observed and the renal pathological changes were detected by HE staining, and the myeloperoxidase activity (MPO) in the renal tissue was detected and Western was used to detect the activity of myeloperoxidase (MPO). The expression of PPAR gamma and Bcl-2 apoptosis related protein in renal tissue was measured by blots. The number of two samples in group designed by t test was tested by SPSS13.0 software package, and the difference of P0.05 was statistically significant.
Results: the creatinine level in CLP group increased gradually with time, and there was no significant difference in 6h creatinine concentration between group CLP and Sham group (P6=0.812, P0.05). The serum creatinine concentration between 12h, 24hCLP and Sham was significantly increased (P12=0.047, P24=0.001, P0.05). There was no significant change in the concentration of 6h between I groups (P6=0.868, P0.05), 12h, 24hCLP+ROSI and Sham+ROSI groups increased significantly (P12=0.033, P24=0.039, P0.05), and there was no significant change in serum creatinine concentration between 0h and 6hCLP+ROSI and Sham+ROSI groups. 0.528, P0.05), compared with the serum creatinine concentration in group 12h and 24hCLP+ROSI, the increase of serum creatinine in group CLP+ROSI was significantly lower than that in group CLP (P12=0.04, P24=0.006, P0.05).6h, 12h, and there was a significant increase in the concentration of serum alpha between the 24hCLP group and the CLP group. The concentration of TNF alpha in serum 6h, 12h and 24h between group CLP+ROSI and group Sham+ROSI was higher (P6=0.00, P12=0.00, P24=0.00, P0.05). 1, P12=0.02, P24=0.01, P0.05), there was no significant difference in myeloperoxidase activity between.0h and 6hCLP+ROSI group and CLP group (P0=0.875, P6=0.13, P0.05), and the level of myeloperoxidase activity in 12h and 24hCLP+ROSI group was significantly lower than that in CLP group. .05) myeloperoxidase activity in group.CLP was significantly higher than that in group Sham (P6=0.00, P12=0.00, P24=0.00, P0.05), and the myeloperoxidase activity in renal tissue between.CLP+ROSI and Sham+ROSI groups was increased (P6=0.00, P12=0.00, P24=0.00,). In group CLP, the symptoms of sepsis and organ dysfunction were found in rats, and the degree of symptom and organ dysfunction in the CLP+ROSI group was less than that of the CLP group. The level of PPAR- gamma and Bcl-2 in each group was measured by Westernblots. The expression of PPAR y and Bcl-2 in the CLP group was significantly lower than that in the CLP+ROSI group.
Conclusion: Rosiglitazone activates PPAR gamma as a ligand, thus alleviates the inflammatory response in sepsis, reduces renal cell apoptosis, improves renal function, reduces renal injury, and has a protective effect on acute renal injury in rats with sepsis.

【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R459.7

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