Neuritin与HSP60在大鼠钝性肝损伤组织的表达规律及意义探讨
发布时间：2018-12-14 13:09
【摘要】：目的:研究大鼠肝损伤后不同时相Neuritin与热休克蛋白60(HSP60)的表达变化,探讨Neuritin与HSP60在大鼠肝损伤修复过程中的作用及意义。方法:将126只S-D大鼠随机分为肝损伤组(A组)、骨折组(B组)和正常组(C组)。A组采用自制BIM-Ⅳ型生物撞击机改良型制作大鼠钝性肝损伤模型,B组制作大鼠右肢股骨截股模型,C组做空白处理。三组分别处理后在6h、12h、1d、3d、5d、7d和14d七个时间点处死大鼠,留取大鼠血清标本及肝组织。分别检测不同时间点大鼠血清ALT、AST浓度以及血清Neuritin、HSP60浓度变化,取肝组织行HE及免疫组化染色,观察肝脏病理学改变及Neuritin、HSP60在大鼠肝损伤组织的时相性表达变化规律。结果:1.A组采用自制生物撞击机成功制作42只大鼠钝性肝损伤模型。2.A组大鼠血清ALT、AST浓度且在6h、12h、1d时高于B组及C组(P0.05),其余时间点三组相比无统计学意义(P0.05)。B组AST浓度在6h、12h、1d时升高,且与A组及C比较均有统计学意义(P0.05)。3.光镜下观察各组大鼠肝脏组织HE染色,A组大鼠肝小叶结构破坏,周边见红色出血带,肝窦内可见大量红细胞,部分肝细胞水肿,局部点状坏死,偶见核分裂象,肝血窦周围可见炎症细胞浸润。随后随着肝损伤的修复,出血明显减少,仍见淋巴细胞及中性粒细胞浸润,内皮细胞明细增多,可见新生毛细血管形成。B组大鼠肝脏肝小叶结构稍紊乱,局部胞质疏松,可见少量炎性细胞。4.免疫组化示,Neuritin蛋白主要表达于胞膜,HSP60蛋白只要表达于胞浆,着色细胞胞浆均呈现为浅黄色和浅棕色,着色肝细胞围绕汇管区分散排列。A组Neuritin及HSP60的表达在各个时间点(6h除外)均显著高于B组及C组(P0.05),且二者表达均呈现先升高后降低的趋势,B组HSP60的表达在12h、1d时高于C组(P0.05),B组Neuritin的表达与C组无明显差异(P0.05)。5.A组血清Neuritin浓度在12h开始升高,3d时达到最高值,后逐渐下降,呈现先升高后降低的趋势;B组大鼠血清Neuritin浓度在1天时逐渐升高,随后一直逐步升高,变化趋势呈现随时间变化逐步升高的变化趋势,各个单独时间点A组、B组及C组三组相比较,各个时间点(6h除外)相比较大鼠血清Neuritin浓度均有统计学意义(P0.05)。A组大鼠血清HSP60在6h开始逐渐升高,12h点达到最高值,后逐渐下降,B组大鼠血清HSP60浓度在6h时逐渐轻微升高,大鼠骨折后12h时最高,随后逐步下降,3d时降至正常浓度,A组大鼠血清HSP60浓度在6h、12h、1d均明显高于B组及C组(P0.05)。结论:Neuritin及HSP60在钝性损伤肝组织中均出现高表达,并且表达随时间变化呈现出一致的变化规律,二者相互作用,可能是促进肝损伤修复的重要因素。
[Abstract]:Aim: to study the expression of Neuritin and heat shock protein 60 (HSP60) in different phases after liver injury in rats, and to explore the role and significance of Neuritin and HSP60 in the repair of liver injury in rats. Methods: 126 S-D rats were randomly divided into three groups: group A (liver injury group), group B (fracture group) and group C (). A group). The femur amputation model of right limb was made in group B and blank treatment was done in group C. The rats in the three groups were killed at 7 days and 14 days after treatment at 6 h, 12 h, 1 day, 1 day, and 5 d, respectively. The serum samples and liver tissue of the rats were collected. The changes of serum ALT,AST concentration and serum Neuritin,HSP60 concentration were detected at different time points respectively. Liver tissues were taken for HE and immunohistochemical staining to observe the pathological changes of liver and the expression of Neuritin,HSP60 in the injured tissues of rats. Results: 1. The serum ALT,AST concentration in group A was significantly higher than that in group B and C (P0.05), and the concentration of serum ALT,AST in group A was significantly higher than that in group B and C (P0.05). There was no significant difference among the three groups at the other time points (P0.05). The AST concentration in the). B group was significantly higher than that in the group A and C (P0.05). HE staining of liver tissue was observed under light microscope. In group A, the structure of hepatic lobule was destroyed, red hemorrhage zone was seen around it, a large number of red blood cells could be seen in hepatic sinuses, some hepatocytes were edematous, local punctate necrosis, and mitosis were occasionally observed. Inflammatory cell infiltration can be seen around the hepatic sinusoids. Subsequently, with the repair of liver injury, the bleeding decreased obviously, the infiltration of lymphocytes and neutrophils, the increase of endothelial cells, and the formation of neocapillaries were observed. In group B, the hepatic lobules were slightly disorganized and the local cytoplasm was loose. A small number of inflammatory cells can be seen. 4. Immunohistochemistry showed that Neuritin protein was mainly expressed in the cell membrane, HSP60 protein was expressed in the cytoplasm, and the cytoplasm of the stained cells was light yellow and light brown. The expression of Neuritin and HSP60 in group A was significantly higher than that in group B and group C (P0.05) at all time points (except 6 h), and the expression of HSP60 in group B was increased first and then decreased. The expression of Neuritin in group A was higher than that in group C (P0.05) at day 1 (P0.05). The concentration of serum Neuritin in group A began to increase at 12h, reached its highest value at 3 days, and then decreased gradually, showing a trend of first rising and then decreasing. The concentration of serum Neuritin in group B gradually increased at 1 day, and then increased gradually. The change trend showed a trend of gradual increase with time, compared with three groups: group A, group B and group C at each single time point. Compared with the larger rats, the serum Neuritin concentration at each time point (except 6 h) was statistically significant (P0.05) in the). A group, the serum HSP60 increased gradually at 6 h, reached the highest value at 12 h, and then decreased gradually. The serum HSP60 level in group B gradually increased slightly at 6h, peaked at 12h after fracture, then decreased gradually, and then decreased to normal concentration at 3d. The serum HSP60 concentration of group A was significantly higher than that of group B and group C (P0.05). Conclusion: both Neuritin and HSP60 are highly expressed in blunt injury liver tissues, and the expression changes with time. The interaction of Neuritin and HSP60 may be an important factor to promote the repair of liver injury.
【学位授予单位】：石河子大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R575
[Abstract]:Aim: to study the expression of Neuritin and heat shock protein 60 (HSP60) in different phases after liver injury in rats, and to explore the role and significance of Neuritin and HSP60 in the repair of liver injury in rats. Methods: 126 S-D rats were randomly divided into three groups: group A (liver injury group), group B (fracture group) and group C (). A group). The femur amputation model of right limb was made in group B and blank treatment was done in group C. The rats in the three groups were killed at 7 days and 14 days after treatment at 6 h, 12 h, 1 day, 1 day, and 5 d, respectively. The serum samples and liver tissue of the rats were collected. The changes of serum ALT,AST concentration and serum Neuritin,HSP60 concentration were detected at different time points respectively. Liver tissues were taken for HE and immunohistochemical staining to observe the pathological changes of liver and the expression of Neuritin,HSP60 in the injured tissues of rats. Results: 1. The serum ALT,AST concentration in group A was significantly higher than that in group B and C (P0.05), and the concentration of serum ALT,AST in group A was significantly higher than that in group B and C (P0.05). There was no significant difference among the three groups at the other time points (P0.05). The AST concentration in the). B group was significantly higher than that in the group A and C (P0.05). HE staining of liver tissue was observed under light microscope. In group A, the structure of hepatic lobule was destroyed, red hemorrhage zone was seen around it, a large number of red blood cells could be seen in hepatic sinuses, some hepatocytes were edematous, local punctate necrosis, and mitosis were occasionally observed. Inflammatory cell infiltration can be seen around the hepatic sinusoids. Subsequently, with the repair of liver injury, the bleeding decreased obviously, the infiltration of lymphocytes and neutrophils, the increase of endothelial cells, and the formation of neocapillaries were observed. In group B, the hepatic lobules were slightly disorganized and the local cytoplasm was loose. A small number of inflammatory cells can be seen. 4. Immunohistochemistry showed that Neuritin protein was mainly expressed in the cell membrane, HSP60 protein was expressed in the cytoplasm, and the cytoplasm of the stained cells was light yellow and light brown. The expression of Neuritin and HSP60 in group A was significantly higher than that in group B and group C (P0.05) at all time points (except 6 h), and the expression of HSP60 in group B was increased first and then decreased. The expression of Neuritin in group A was higher than that in group C (P0.05) at day 1 (P0.05). The concentration of serum Neuritin in group A began to increase at 12h, reached its highest value at 3 days, and then decreased gradually, showing a trend of first rising and then decreasing. The concentration of serum Neuritin in group B gradually increased at 1 day, and then increased gradually. The change trend showed a trend of gradual increase with time, compared with three groups: group A, group B and group C at each single time point. Compared with the larger rats, the serum Neuritin concentration at each time point (except 6 h) was statistically significant (P0.05) in the). A group, the serum HSP60 increased gradually at 6 h, reached the highest value at 12 h, and then decreased gradually. The serum HSP60 level in group B gradually increased slightly at 6h, peaked at 12h after fracture, then decreased gradually, and then decreased to normal concentration at 3d. The serum HSP60 concentration of group A was significantly higher than that of group B and group C (P0.05). Conclusion: both Neuritin and HSP60 are highly expressed in blunt injury liver tissues, and the expression changes with time. The interaction of Neuritin and HSP60 may be an important factor to promote the repair of liver injury.
【学位授予单位】：石河子大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R575
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