高活性小牛血去蛋白提取物对化学性肝损伤的保护作用及其机制研究

发布时间：2018-06-20 12:05

本文选题：小牛血去蛋白提取物 + 四氯化碳　；参考：《北华大学》2017年硕士论文

【摘要】：目的:观察小牛血去蛋白提取物(deproteinized extract of calf blood,DECB)对化学性肝损伤小鼠的保护作用,并初步探讨其作用机制。方法:建立两种小鼠急性肝损伤模型,分别为酒精致急性肝损伤模型和CCl4致急性肝损伤模型。酒精性肝损伤实验,取健康ICR种小鼠60只,随机分组法分为:空白组、模型组、阳性药物组和高活性小牛血去蛋白提取物低、中、高剂量组,每组10只。灌胃给药,空白组给予生理盐水20 m L·kg-1,DECB低、中和高剂量组分别给予DECB 0.25、0.50、1.00 g·kg-1,阳性药物组给予护肝片0.63g·kg-1。每天1次,连续30d,末次给药1 h后,除空白组外,其他各组一次性给予50%乙醇14 m L·kg-1,并禁食16 h建立急性酒精性肝损伤模型。测定小鼠的耐受酒精时间和醉酒时间,检测血清中丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)活性,检测肝组织中甘油三酯(TG)、谷胱甘肽(GSH)和丙二醛(MDA)水平,并进行肝组织病理切片检查。CCl4致急性肝损伤实验,取健康ICR种小鼠40只,随机分组法分为空白组、模型组、阳性药物组和DECB组每组10只。各组小鼠灌胃给药,空白组给予生理盐水20 m L·kg-1,DECB组给予DECB 1.00g·kg-1,阳性药物组给予护肝片0.63g·kg-1。每天1次,连续30d;末次给药2 h后,对照组小鼠腹腔注射调和油溶液,其余各组小鼠腹腔注射10%CCl4调和油溶液(10 m L·kg-1)并禁食,建立CCl4致急性肝损伤模型。测定各组小鼠血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)活性和肝组织总超氧化物歧化酶(T-SOD)、谷胱甘肽(GSH)及丙二醛(MDA)水平;HE染色法观察肝组织病理改变,TUNEL方法检测肝组织细胞凋亡,通过m RNA表达芯片筛选DECB组和模型组的差异表达基因及其相关通路。结果:酒精性肝损伤实验发现,与模型组比较,DECB组明显减轻小鼠醉酒症状,DECB高剂量组和阳性药组小鼠酒精耐受时间延长(P0.05㖞,醉酒时间缩短(P0.05㖞;DECB中、高剂量组小鼠血清ALT和AST活性均明显低于模型组(P0.05㖞;与模型组比较,DECB中、高剂量组和阳性药物组小鼠肝组织中MDA和TG水平明显降低,差异有统计学意义(P0.05㖞;与模型组比较,DECB高剂量组小鼠乙醇所致肝组织病理学损伤明显减轻。CCl4致急性肝损伤实验发现DECB组小鼠血清ALT和AST活性均明显低于模型组(P0.05㖞;与模型组比较,DECB组小鼠肝组织MDA和GSH水平明显降低(P0.05㖞,T-SOD水平明显升高(P0.05㖞;肝组织病理学损伤明显减轻,凋亡指数降低(P0.05㖞;在聚类结果中发现细胞凋亡通路中共有9个差异表达基因,2个基因表达上调,7个基因表达下调。结论:DECB对化学性肝损伤具有保护作用,可以增强肝脏抗氧化能力,抑制肝细胞凋亡,其机制可能与调控细胞凋亡通路基因表达有关。
[Abstract]:Objective: to observe the protective effect of deproteinized extract of calf blood extract (DECB) on chemically induced liver injury in mice, and to explore its mechanism. Methods: two kinds of acute liver injury models were established in mice, which were acute liver injury induced by alcohol and CCL _ 4, respectively. In alcoholic liver injury experiment, 60 ICR mice were randomly divided into blank group, model group, positive drug group and high active calf blood deproteinization extract with 10 rats in each group. The normal saline 20 mL kg -1 DECB was given to the blank group, the middle and high dose groups were given the DECB 0.25 ~ 0.50 ~ 0.50 g 路kg ~ (-1) respectively, and the positive drug group was given 0.63 g 路kg ~ (-1) of protective liver tablet. Once a day for 30 days, after the last administration for 1 hour, the other groups were given 50% ethanol 14ml 路kg -1 for one time, and fasting for 16 h to establish the model of acute alcoholic liver injury. The alcohol tolerance time and drunkenness time of mice were measured. The activities of alanine aminotransferase (alt) and aspartate aminotransferase (AST) in serum and the levels of triglyceride, glutathione (GSH) and malondialdehyde (MDAs) in liver tissue were measured. 40 ICR mice were randomly divided into blank group, model group, positive drug group and DECB group. Mice in each group were given intragastric administration of DECB 1.00 g kg-1in the blank group and 0.63 g kg-1 in the positive drug group. The control group was given normal saline 20 mL 路kg ~ (-1) 路kg ~ (-1) and the positive drug group was given 0.63 g / kg ~ (-1). After the last administration for 2 hours, the control mice were injected with the blending oil solution intraperitoneally, and the other mice were injected intraperitoneally with the 10l4 blend oil solution 10 mL kg-1) and fasted to establish the model of acute liver injury induced by CCl4. The activity of serum alanine aminotransferase (alt), aspartate aminotransferase (AST), total superoxide dismutase (T-SOD), glutathione (GSH) and malondialdehyde (MDA) were measured in each group of mice. Detection of apoptosis in liver tissue, The differentially expressed genes and their related pathways in DECB group and model group were screened by mRNA expression microarray. Results: compared with the model group, the alcoholic liver injury test showed that the alcohol tolerance time of the DECB group and the positive drug group were significantly reduced compared with the model group, and the alcohol tolerance time was prolonged in the DECB group and the positive drug group, and the alcohol tolerance time was shortened in the DECB group, the alcohol tolerance time was shortened in the DECB group, and the alcohol tolerance time was shortened in the DECB group. The activities of alt and AST in high dose group were significantly lower than those in model group (P 0.05), and the levels of MDA and TG in liver tissue of mice in high dose group and positive drug group were significantly lower than those in model group. Compared with the model group, the liver histopathological damage induced by ethanol in the DECB group was significantly reduced. The serum alt and AST activities in the DECB group were significantly lower than those in the model group, and those in the DECB group were significantly lower than those in the model group. Compared with the model group, the levels of MDA and GSH in the liver tissue of the DECB group decreased significantly, the level of P0.05TSOD increased significantly, and the pathological damage of the liver tissue was alleviated obviously. The results of cluster analysis showed that there were 9 differentially expressed genes, 2 up-regulated genes and 7 down-regulated genes in the apoptotic pathway. Conclusion DECB has protective effect on chemically induced liver injury. It can enhance the antioxidant ability of liver and inhibit the apoptosis of hepatocytes. The mechanism may be related to the regulation of gene expression of apoptosis pathway.
【学位授予单位】：北华大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R965

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