Neuritin與HSP60在大鼠鈍性肝損傷組織的表達(dá)規(guī)律及意義探討
發(fā)布時(shí)間：2018-12-14 13:09
【摘要】：目的:研究大鼠肝損傷后不同時(shí)相Neuritin與熱休克蛋白60(HSP60)的表達(dá)變化,探討Neuritin與HSP60在大鼠肝損傷修復(fù)過(guò)程中的作用及意義。方法:將126只S-D大鼠隨機(jī)分為肝損傷組(A組)、骨折組(B組)和正常組(C組)。A組采用自制BIM-Ⅳ型生物撞擊機(jī)改良型制作大鼠鈍性肝損傷模型,B組制作大鼠右肢股骨截股模型,C組做空白處理。三組分別處理后在6h、12h、1d、3d、5d、7d和14d七個(gè)時(shí)間點(diǎn)處死大鼠,留取大鼠血清標(biāo)本及肝組織。分別檢測(cè)不同時(shí)間點(diǎn)大鼠血清ALT、AST濃度以及血清Neuritin、HSP60濃度變化,取肝組織行HE及免疫組化染色,觀察肝臟病理學(xué)改變及Neuritin、HSP60在大鼠肝損傷組織的時(shí)相性表達(dá)變化規(guī)律。結(jié)果:1.A組采用自制生物撞擊機(jī)成功制作42只大鼠鈍性肝損傷模型。2.A組大鼠血清ALT、AST濃度且在6h、12h、1d時(shí)高于B組及C組(P0.05),其余時(shí)間點(diǎn)三組相比無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。B組AST濃度在6h、12h、1d時(shí)升高,且與A組及C比較均有統(tǒng)計(jì)學(xué)意義(P0.05)。3.光鏡下觀察各組大鼠肝臟組織HE染色,A組大鼠肝小葉結(jié)構(gòu)破壞,周邊見(jiàn)紅色出血帶,肝竇內(nèi)可見(jiàn)大量紅細(xì)胞,部分肝細(xì)胞水腫,局部點(diǎn)狀壞死,偶見(jiàn)核分裂象,肝血竇周?chē)梢?jiàn)炎癥細(xì)胞浸潤(rùn)。隨后隨著肝損傷的修復(fù),出血明顯減少,仍見(jiàn)淋巴細(xì)胞及中性粒細(xì)胞浸潤(rùn),內(nèi)皮細(xì)胞明細(xì)增多,可見(jiàn)新生毛細(xì)血管形成。B組大鼠肝臟肝小葉結(jié)構(gòu)稍紊亂,局部胞質(zhì)疏松,可見(jiàn)少量炎性細(xì)胞。4.免疫組化示,Neuritin蛋白主要表達(dá)于胞膜,HSP60蛋白只要表達(dá)于胞漿,著色細(xì)胞胞漿均呈現(xiàn)為淺黃色和淺棕色,著色肝細(xì)胞圍繞匯管區(qū)分散排列。A組Neuritin及HSP60的表達(dá)在各個(gè)時(shí)間點(diǎn)(6h除外)均顯著高于B組及C組(P0.05),且二者表達(dá)均呈現(xiàn)先升高后降低的趨勢(shì),B組HSP60的表達(dá)在12h、1d時(shí)高于C組(P0.05),B組Neuritin的表達(dá)與C組無(wú)明顯差異(P0.05)。5.A組血清Neuritin濃度在12h開(kāi)始升高,3d時(shí)達(dá)到最高值,后逐漸下降,呈現(xiàn)先升高后降低的趨勢(shì);B組大鼠血清Neuritin濃度在1天時(shí)逐漸升高,隨后一直逐步升高,變化趨勢(shì)呈現(xiàn)隨時(shí)間變化逐步升高的變化趨勢(shì),各個(gè)單獨(dú)時(shí)間點(diǎn)A組、B組及C組三組相比較,各個(gè)時(shí)間點(diǎn)(6h除外)相比較大鼠血清Neuritin濃度均有統(tǒng)計(jì)學(xué)意義(P0.05)。A組大鼠血清HSP60在6h開(kāi)始逐漸升高,12h點(diǎn)達(dá)到最高值,后逐漸下降,B組大鼠血清HSP60濃度在6h時(shí)逐漸輕微升高,大鼠骨折后12h時(shí)最高,隨后逐步下降,3d時(shí)降至正常濃度,A組大鼠血清HSP60濃度在6h、12h、1d均明顯高于B組及C組(P0.05)。結(jié)論:Neuritin及HSP60在鈍性損傷肝組織中均出現(xiàn)高表達(dá),并且表達(dá)隨時(shí)間變化呈現(xiàn)出一致的變化規(guī)律,二者相互作用,可能是促進(jìn)肝損傷修復(fù)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.
【學(xué)位授予單位】：石河子大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：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.
【學(xué)位授予單位】：石河子大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R575
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