【作者】 彭颖琼；

【导师】 鲁猛厚；

【作者基本信息】 中南大学 ， 内科学， 2010， 博士

【摘要】 目的：构建脓毒症大鼠实验模型,观察重组人生长激素(rhGH)对脓毒症大鼠的保护作用。方法：SD大鼠75只,随机分为3组(每组25只)：A组为对照组,仅予以开腹游离盲肠,B组为脓毒症组,开腹游离盲肠并行盲肠结扎穿孔术制造脓毒症模型,C组为rhGH治疗组,行盲肠结扎穿孔模型的同时予以皮下注射rhGH1.0U/kg一次,0h、2h、8h、12h、24h后观察大鼠血清谷丙转氨酶(ALT)活性、TNF-α水平和死亡率,24h后取心脏血行血培养。比较不同时间点ALT活性、TNF-α表达、死亡率和24h血培养结果。结果：A组未见大鼠死亡,B组24h存活率48%(12／25)、C组24h存活率76%(19／25)。与A组相比,B、C组大鼠血清中ALT、TNF-α表达、血培养阳性率明显增加(P<0.05),而与B组相比,C组大鼠血清中ALT、TNF-α表达、血培养阳性率明显降低(P<0.05)。结论：1、成功构建盲肠结扎穿孔的大鼠实验性脓毒症模型。2、重组人生长激素对盲肠结扎穿孔诱导的大鼠脓毒症具有明显的保护作用。目的：观察重组人生长激素对盲肠结扎穿孔诱导的脓毒症大鼠肝脏的影响,并初步探讨其作用机制。方法：实验动物分组、脓毒症模型的构建及实验动物给药同第一部分,real-time PCR测定肝组织中TNF-αmRNA和HMGB1 mRNA的表达,Western blot测定肝组织中HMGB1和活化Caspase-3的蛋白表达,免疫组化测定肝组织中NF-κB的表达,原位凋亡检测TUNEL法检测肝细胞的凋亡情况。结果：与A组相比,B、C组大鼠肝组织中TNF-αmRNA、HMGB1 mRNA、HMGB1和活化Caspase-3表达增加,NF-κB核移位及肝细胞凋亡增加(P<0.05)。与B组相比,rhGH可以明显降低C组大鼠肝组织中TNF-αmRNA、HMGB1 mRNA、HMGB1和活化Caspase-3的表达,减少NF-κB核移位及肝细胞凋亡(P<0.05)。结论：rhGH通过抑制大鼠肝组织NF-κB活化,下调TNF-α、HMGB1表达,降低活化Caspase-3蛋白表达及肝细胞的凋亡,对肝细胞产生一定保护作用。更多还原

【Abstract】 Objective:To investigate the protective effect of recombinant human growth hormone(rhGH) on sepsis in rats.Methods:seventy-five SD rars were randomly divided into 3 groups, sham operation group(group A), sepsis group(group B) and rhGH group(group C). Cecal ligation and puncture operation were done in sepsis and rhGH group. In addition, rhGH(1u/kg) was injected subcutaneously at the same time point operation performed in group C. Death rate of each group at 24 hours after operation was caculated and survived rats were killed in order to get blood and liver samples. Activity of ALT was tested and level of TNF-a was detected by ELISA. Bacteria translocation were detected by blood culture.Results:The survival rate in 24 hours was 100% in group A,48% in group B and 76% in group C. The activity of ALT, lever of TNF-a and positive rate of blood culture in group B and C were significantly higher than that in group A(P<0.05). The activity of ALT, lever of TNF-a and positive rate of blood culture in group C were significantly lower than that in group B(P<0.05).Conclusion:The Cecal ligation and puncture operation induced sepsis in rats were established successfully. Recombinant human growth hormone had an obviously protective effect on Cecal ligation and puncture operation induced sepsis rats. Objective:To investigate the mechanism of recombinant human growth hormone on sepsis in rats liver.Methods:seventy-five SD rars were randomly divided into 3 groups, sham operation group(group A), sepsis group(group B) and rhGH group(group C). Cecal ligation and puncture operation were done in sepsis and rhGH group. In addition, rhGH(1u/kg) was injected subcutaneously at the same time point operation performed in group C. Survived rats were killed in order to get liver samples at 24 hours after operation. The expression of TNF-a mRNA and HMGB1 mRNA were tested by real-time PCR. The expression of HMGB1 and active Caspase-3 were detected by western blot. Nucleal positive rate of NF-KBp65 protein in liver tissue was determined by immunohistochemical staining. Apoptosis of liver cell was measured by Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL).Results:The expression of TNF-a mRNA, HMGB1 and active Caspase-3 in group B and C were significantly higher than that in group A(P<0.05). Nucleal positive rate of NF-κBp65 protein in liver tissue and apoptosis of liver cell in group B and C were markedly higher than that in group A(P<0.05). The expression of TNF-a mRNA, HMGB1 and active Caspase-3 in group C were greatly lower than that in group B(P<0.05). Nucleal positive rate of NF-KBp65 protein in liver tissue and apoptosis of liver cell in group C were prominently lower than that in group B(P<0.05).Conclusion:Recombinant human growth hormone can protect the liver by decreasing the apoptosis of liver cell in sepsis, inhibiting the production and release of inflammatory factors such as TNF-αand HMGB1, inhibiting activation of NF-κB in liver.更多还原