李梁和,陈忠胜,田甜,郑璐,李珀,詹玮
LI Lianghe,CHEN Zhongsheng,TIAN Tian,ZHENG Lu,LI Po,ZHAN Wei

• 1:贵州医科大学外科学教研室
• 2:贵州医科大学
• 3:贵州医科大学病理生理学教研室
• 4:贵州医科大学附院病理科

摘要(Abstract)：

目的:研究蓝莓对四氯化碳(CCl_4)致肝损伤大鼠肝功能、肝纤维化及组蛋白乙酰化修饰的影响。方法:将50只SD大鼠随机均分为对照组、模型组、蓝莓治疗组、蓝莓干预组和自然恢复组5组;模型组、蓝莓治疗组、蓝莓干预组和自然恢复组均给予CCl_4植物油溶液皮下注射4周处死;蓝莓治疗组于造模结束后给予蓝莓汁灌胃8周,共12周;蓝莓干预组在造模4周前至造模结束均以蓝莓汁灌胃,共8周;自然恢复组在造模4周后自然饮食8周,共12周;检测5组大鼠血清中肝功能[丙氨酰胺基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)]2项指标及肝纤维化[血清透明质酸(HA)、Ⅳ型胶原(CⅣ)、血清层粘连蛋白(LN)、Ⅲ型前胶原(PⅢNP)]4项指标,肝脏切片HE染色行肝脏病理检查,肝脏组织进行核浆分离后检测各组大鼠肝脏组织中组蛋白乙酰化acH3K9、acH3K14和acH3K18蛋白的表达。结果:与对照组比较,各组大鼠血清中ALT、AST、HA和CⅣ明显增高(P<0.05);与模型组比较,蓝莓治疗组及蓝莓干预组大鼠血清ALT、AST、HA和CⅣ水平均明显下降(P<0.05);肝脏HE染色显示,4周CCl_4植物油溶液皮下注射造模成功;与模型组比较,蓝莓干预组大鼠肝脏有明显改善,而蓝莓治疗组及自然恢复组大鼠的肝脏有轻微改善;与对照组比较,蓝莓治疗组及蓝莓干预组大鼠肝脏组织中acH3K9、acH3K14和acH3K18修饰水平的差异不明显(P>0.05);与模型组比较,余各组大鼠肝脏组织中acH3K9、acH3K14和acH3K18蛋白修饰水平增高(P<0.01);与自然恢复组比较,蓝莓治疗组和蓝莓干预组肝组织中acH3K9、acH3K14、acH3K18修饰降低程度比模型组稍高(P<0.05)。结论:灌胃蓝莓可改善CCl_4肝损伤模型大鼠的部分肝功能指标,其机制可能与其影响大鼠肝组织组蛋白乙酰化修饰水平有关。
Objective: To study the effects of blueberry on liver function,fibrosis and histone acetylation in rats with liver injury induced by carbon tetrachloride. Methods: 50 SD rats were randomly divided into 5 groups: the control group,the model group,the blueberry treatment group,the blueberry intervention group and the natural recovery group. The model group,blueberry treatment group,blueberry intervention group and natural recovery group were given the carbon tetrachloride oil solution subcutaneously for 4 weeks. Then the blueberry treatment group was treated with blueberry juice by gavage for 8 weeks after the subcutaneous injection,for a total of 12 weeks; the blueberry intervention group was treated with blueberry juice by gavage since 4 weeks before the subcutaneous injection until the injection ended,for a total of 8 weeks,and the natural recovery group had a natural diet for 8 weeks 4 weeks after the subcutaneous injection,for a total of 12 weeks. Two indexes of liver function in serum[alanine aminotransferase( ALT),aspartate aminotransferase( AST) ]and four indexes of hepatic fibrosis [serum hyaluronic acid( HA),type Ⅳcollagen( CⅣ),serum laminin( LN),type Ⅲ procollagen] were measured in the 5 groups of rats. Liver pathological examination was performed with HE staining in liver slices. The liver tissue was separated by nuclear plasma separation to detect the expression of acH3K9,acH3K14 and acH3K18 protein in the liver tissues of each group. Results: Compared with the control group,the serum levels of ALT,AST,HA and C IV increased significantly( P < 0. 05). Compared with the model group,the serum levels of ALT,AST,HA and C IV in the blueberry treatment group and the blueberry intervention group significantly decreased( P < 0. 05).The liver HE staining showed that the four week's subcutaneous injection of carbon tetrachloride oil solution was successful. Compared with the model group,the liver improvement of the rats in the blueberry intervention group was obvious,but the liver improvements of the blueberry treatment group and the natural recovery group were slight. Compared with the control group,the acH3K9,acH3K14 and acH3K18 modification levels in the liver tissues of the blueberry and blueberry intervention group were not significantly different( P > 0. 05) and the model group. Ac H3K9,acH3K14 and acH3K18 protein modification levels in the liver tissues of the remaining groups increased( P < 0. 01). Compared with the natural recovery group,the reduction of modification levels of acH3K9,acH3K14 and acH3K18 in the liver tissues of the blueberry and blueberry intervention group were slightly higher than that in the model group( P < 0. 05). Conclusion: Treatment of blueberry by gavage can improve partial liver function indexes in rats with liver injury induced by carbon tetrachloride,and the mechanism may be related to its effects on the modification level of histone acetylation in liver tissues of rats.

关键词(KeyWords)： 大鼠,Sprague-Dawley;肝功能实验;肝纤维化;蓝莓;四氯化碳;组蛋白乙酰化
rats,Sprague-Dawley;liver function experiment;hepatic fibrosis;blueberry;carbon tetrachloride;histone acetylation

Abstract:

Keywords:

基金项目(Foundation): 贵州省卫生计生委科学技术基金项目(gzwjkj2018-1-035);; 贵阳市科技计划项目[筑科合同(20161001)052号]

作者(Author): 李梁和,陈忠胜,田甜,郑璐,李珀,詹玮
LI Lianghe,CHEN Zhongsheng,TIAN Tian,ZHENG Lu,LI Po,ZHAN Wei

DOI: 10.19367/j.cnki.1000-2707.2018.07.005

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