摘要
目的探讨海兔素(aplysin)对酒精性肝损伤大鼠免疫功能的影响。方法雄性wistar大鼠50只,随机分为正常对照组(每日给予生理盐水灌胃1次)、酒精模型组(每日给予50%乙醇8 ml/(kg bw·d)灌胃,2 w后酒精剂量增至12 ml/(kg bw·d),继续灌胃4 w;)、海兔素低、中、高剂量组(海兔素低中高剂量组每日分别给予Aplysin50,、100、150 mg/(kg bw·d)灌胃,1 h后灌酒精,剂量同模型组n=10)。实验共6w。末次灌胃12 h后,称重后给予7%水合氯醛麻醉,腹主动脉取血,留取肝脏、脾脏并计算脾指数;HE染色观察肝脏组织的病理学改变;采用酶联免疫吸附(ELISA)法检测血清中白介素-2(IL-2)、白介素-10(IL-10)的含量,并计算IL-2/IL-10比值;免疫组化法检测肝组织中CD4+、CD8+T淋巴细胞亚群表达,计数CD4+T细胞和CD8+T细胞,并计算CD4+/CD8+比值。结果海兔素中、高剂量组的脾指数分别为(0.205±0.173),(0.196±0.189),均低于酒精模型组(2.765±0.094),差异有统计学意义(P<0.05)。HE染色观察结果,酒精模型组大鼠肝小叶结构模糊,细胞排列紊乱,胞浆空泡化,肝细胞肿大,出现明显的脂肪变性,可见炎性细胞浸润;海兔素各剂量组肝小叶结构也有不同程度的破坏,与酒精模型组相比,肝细胞坏死程度明显减轻。模型组血清中IL-2含量与正常对照组相比显著升高,IL-10含量降低,IL-2/IL-10比值升高,差异均有统计学意义(P<0.05);海兔素低、中、高剂量组与模型组相比IL-2含量降低,IL-10含量升高,IL-2/IL-10比值降低,差异均有统计学意义(P<0.05)。模型组与正常对照组相比CD4+表达升高,CD8+表达降低,CD4+/CD8+比值升高,差异均有统计学意义(P<0.05);海兔素各剂量组与模型组相比CD4+表达降低,CD8+表达升高,CD4+/CD8+比值降低,差异均有显著性(P<0.05)。其中,随着海兔素剂量的增加,IL-2、CD4+表达逐渐降低,且呈剂量依赖关系,各剂量组间差异均有统计学意义(P<0.05);CD8+表达逐渐升高,低剂量组与中、高剂量组间差异有统计学意义(P<0.05);IL2/IL-10比值逐渐降低,低、高剂量组间差异有统计学意义(P<0.05)。结论海兔素对酒精性肝损伤大鼠具有一定的免疫调节作用。
Objective This study aimed to explore the immunomodulation effect of aplysin in rats with alcohol- induced hepatic injury. Methods Fifty male Wistar rats were randomly divided into five groups of equal size： normal control group （normal saline once a day）, low-, medium-, and high-dose aplysin groups （50, 100, 150 mg/（kg bw · d aplysin） and alcohol-model group. Except normal control group, all groups were administrated orally with 8 ml/kg· d of 50 % （v/v） ethanol for 2 weeks followed by an increasing intake of ethanol up to 12 ml/（kg.d） for the remaining 4 weeks. The experiment lasted for 6 weeks. Twelve hours after the last ethanol treatment, the rats were anesthetized. Then, blood and spleen samples were collected. The liver was obtained for histopathology examination by HE staining. The levels of serum IL-2 and IL-10 were measured by ELISA and the ratio of IL-2/IL-10 was calculated.The expression of CD4＋, CD8＋T lymphocytes in liver was detected by immunosorbent assay and the ratio of CD4＋/CD8＋ was calculated. Results In medium- and high-dose aplysin treatment groups, the spleen index was 0.205±0.173 and 0.196±0.189 respectively, which was significantly lower than that of alcohol model group （P〈0.05）. Pathological observation of liver tissue by HE staining showed the hepatic lobule structure was fuzzy; the arrangement of cells was disordered; liver cells were swelling; there was a significant steatosis and inflammatory cell infiltration in alcohol model group. From hepatic lobule structure, there was different degree of damage in each aplysin-treated group. In addition, hepatocyte necrosis was significantly reduced in aplysin treated groups compared with that in aocohol model group. Compared with normal control group, the level of serum IL-2 in alcohol model group was significantly increased, while the level of serum IL-10 was reduced and the ratio of IL2/IL-10 was increased （P〈0.05）. Compared with alcohol model group, the level of serum IL-2 was reduced and the level of IL-10 was increased in each aplysin treatment group. The ratio of IL-2/IL-10 was reduced （P 〈 0.05）. Compared with normal control group, the expression of CD4＋ was increased, CD8＋ was reduced, and the ratio of CD4＋/CD8＋ was increased （P〈0.05） in alcohol model group. However, compared with alcohol model group, the expression of CD4＋ was reduced, CD8＋ was increased and the ratio of CD4＋/CD8＋ was reduced （P〈0.05） in each aplysin treatment group. With the increase of aplysin dose, the level of IL-2 and the expression of CD4＋ were gradually reduced. Additionally, the difference of IL-2 and CD4 among each dose group had a statistical significance （P〈0.05）. The expression of CD8＋ was gradually increased and difference between low dose and medium or high dose aplysin group was statistically significant （P〈0.05）. Furthermore, the ratio of IL2/IL-10 was gradually reduced and difference between low and high dose group was statistically significant （P〈0.05）. Conclusion Aplysin has a certain immunomodulatory effect in rats with alcohol-induced hepatic injury.
出处
《营养学报》
CAS
CSCD
北大核心
2015年第5期484-489,共6页
Acta Nutrimenta Sinica
基金
国家"十二五"科技支撑计划项目(2012BAD33B01)
国家自然科学基金(31171671)资助
关键词
酒精性肝病
alcoholic liver disease
aplysin
immunoregulation
interleukin-2/interleukin-10
CD4＋/CD8＋
