肝炎肝郁脾虚证动物模型综合药效评价指标的建立

发布时间：2018-05-01 08:15

本文选题：肝郁脾虚证 + 药效评价指标　；参考：《延边大学》2012年硕士论文

【摘要】：目的：通过皮下注射四氯化碳、慢性束缚尝试建立肝郁脾虚证模型,并用经典治疗药逍遥丸化裁方对该模型进行干预,初步形成慢性肝炎肝郁脾虚证的建立方法,形成综合药效评价指标。方法：成年雄性大鼠32只,体重250g-300g。购入大鼠后适应性饲养一周后,分为A(空白组),B(模型组),C(逍遥丸化裁方组)三组。空白组8只其余每组12只,除A组大鼠外,其余24只大鼠均按0.5ml/100g体重皮下注射10%CCL4豆油溶液,首次剂量加倍,每周注射2次,共四周。第四周从模型组中任选两只大鼠测ALT、AST来确定肝损伤的程度。开始造模一周后,以慢性束缚方法制作慢性应激大鼠模型,将B、C组大鼠束缚于束缚筒内,放入饲养箱中,每日3h,早8点到11点,连续21d。A组不予束缚,但于相同时间点禁食,放置于各自饲养箱中3h/d,连续21d。C组大鼠在实验开始1周后,每次开始束缚前30mmin给各组大鼠灌胃给药,1次/d,共用3周。(以人用药量的30倍作为大鼠给药的逍遥丸化裁方组,灌胃容积为20mL/kg体重)。A组则分别灌服同等换算体积的蒸馏水。观察实验动物的情绪行为、日饮用量、大便溏薄观察、自主活动能力。给药后每周灌服D-木糖接取尿液检测尿液中D-木糖排泄率。检测血清中5-羟色胺(5-HT)、去甲肾上腺素(NE)、多巴胺(DA)水平,丙氨酸氨基转移酶(ALT)和门冬氨酸氨基转移酶(AST)的水平,血浆生长抑素(SS)、胃动素(MTL)、血管活性肠肽(VIP)的水平,总胆红素(TBILI)、直接胆红素(DBILI)、总蛋白(TP)、白蛋白(ALB)的变化,白介素-2、6、8的水平。观察肝脏病理变化。所有指标进行组间t检验统计学处理。结果：行为学指标：实验动物出现便溏、饮食饮水量减少、暴躁易怒、体重降低等症状。逍遥丸化裁方组与模型组比较,肝脏、脾脏、肾上腺指数降低。生化指标显示与模型组比较：1.空白组、逍遥丸化裁方组尿液中D-木糖的含量低于模型组。2.5-HT含量升高(P0.05),DA明显升高(P0.01),NE含量又降低趋势。3.血清中ALT和AST的水平明显低于模型组(P0.05)。4.ALB明显升高(P0.001)、DBLI有降低趋势、TP明显升高(P0.05),TBILl各组间无明显差异。5.MTL明显升高(P0.05)、VIP显著升高(P0.001)、SS升高(P0.05)。 6.IL-2降低(P0.05)、IL-6降低、IL-8降低。7.空白组肝组织结构正常,模型组肝小叶结构不清,可见明显的片状坏死,伴有炎症细胞浸润。逍遥丸组肝小叶结构尚正常,可见明显的混浊肿胀气球样变或脂肪变性,散在点状坏死。结论：肝炎肝郁脾虚证动物模型综合药效评价指标的建立。
[Abstract]:Objective: to establish the model of liver stagnation and spleen deficiency by subcutaneous injection of carbon tetrachloride (CCL 4), and interfere with Xiaoyao Pill (Xiaoyao Pill), a classical therapeutic drug, in order to form the method of establishing liver stagnation and spleen deficiency syndrome in chronic hepatitis. A comprehensive evaluation index of drug efficacy was formed. Methods: 32 adult male rats were weight 250 g-300 g. After a week of adaptive feeding, rats were divided into three groups: group A (model group). Except for group A, the other 24 rats were subcutaneously injected with 10L4 soybean oil solution according to the body weight of 0.5ml/100g. The first dose was doubled, twice a week, for a total of four weeks. In the fourth week, two rats were selected from the model group to determine the degree of liver injury. One week after the model was established, the chronic stress rat model was made by chronic restraint method. The rats of group C were tied to the restraint tube and put into the feeding box for 3 hours daily, 8: 00 to 11:00 in the morning. The rats in the 21d.A group were not bound, but fasted at the same time. The rats in the 21d.C group were placed in their respective feeding boxes for 3 h / d. The rats in the 21d.C group were given intragastric administration of 30mmin for 3 weeks after one week of experiment and before each group began to be bound. (the group of Xiaoyao pills treated with Xiaoyao Pill, 30 times of the dosage of human medicine) was given distilled water with the same volume of distilled water in the group with 20mL/kg body weight. Observe the emotional behavior of experimental animals, daily consumption, loose stool observation, independent activity. The excretion rate of D-xylose in urine was determined by weekly administration of D-xylose. The levels of serotonin 5-HT, norepinephrine (NE), dopamine (DA), alanine aminotransferase (alt) and aspartate aminotransferase (AST), plasma somatostatin (SS), motilin (MTL), vasoactive intestinal peptide (VIP) were measured. The changes of total bilirubin (TBILI), direct bilirubin (DBILI), total protein (TPN), albumin (ALB), and the level of interleukin-2 (TBILI). Liver pathological changes were observed. All the indexes were analyzed by t-test. Results: behavioral indicators: loose stools, decreased diet, irritability, weight loss and so on. Compared with model group, the index of liver, spleen and adrenal gland decreased in Xiaoyao pill group. The biochemical index display was compared with that of the model group. In blank group, the content of Dxylose in urine of Xiaoyao pill group was lower than that of model group. The serum levels of ALT and AST were significantly lower than those of the model group (P 0.05). 4. The levels of ALT and AST in serum were significantly lower than those in the model group. There was a tendency to decrease the level of P0.001TBI. There was no significant difference between the three groups. 5. The levels of ALT and AST were significantly higher than those of the model group. 6.IL-2 decreased P0.05, IL-6 decreased, IL-8 decreased. In blank group, the structure of liver tissue was normal, the structure of hepatic lobule in model group was not clear, obvious flake necrosis and inflammatory cell infiltration could be seen. In Xiaoyao Pill group, the structure of hepatic lobule was normal, obvious turbid swelling, balloon degeneration or steatosis, scattered necrosis. Conclusion: the animal model of liver stagnation and spleen deficiency syndrome was established.
【学位授予单位】：延边大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R-332

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