丹参多糖对小鼠免疫性肝损伤中炎症因子的调控
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摘要
将60只昆明小鼠随机分为正常对照组,LPS模型组,丹参多糖高、中、低(500,250,125mg/kg)保护组,每组12只。保护组用相应剂量的丹参多糖连续灌胃7d,1次/d。模型组和正常组给以等剂量的生理盐水。模型组和保护组在末次灌胃1h后,腹腔注射LPS(10mg/kg)建立肝损伤模型;正常对照组腹腔注射等体积的生理盐水。各组小鼠于注射LPS 2h后,眼球取血分离血清,测定其血清谷丙转氨酶(AST)、谷草转氨酶(ALT)的活性和TNF-α、INF-γ、IL-6、IL-10含量;HE染色观察肝组织形态学变化;RT-PCR检测肝组织中TNF-α、INF-γmRNA的表达。结果显示,与正常对照组相比,LPS模型组小鼠血清AST、ALT活性和TNF-α、INF-γ、IL-6、IL-10含量均显著增加(P<0.05或P<0.01)。丹参多糖各保护组(500,250,125mg/kg)均可显著降低AST、ALT的活性和TNF-α、INF-γ、IL-6、IL-10的含量,并可减少肝组织中TNF-α、INF-γmRNA的表达。且丹参多糖对肝损伤的这种保护作用呈现一定的剂量依赖性。结果表明,丹参多糖对免疫性肝损伤具有显著的保护作用,且这种保护作用可能与丹参多糖降低炎症因子水平有关。
The aim of the present study was to study the protective effect of salvia miltiorrhiza polysaccharides on immunological liver injury induced by LPS,and to explore a possible mechanism.60 Kunming mouse were randomly divided into 5 groups:the normal group,the model group,the different does of salvia miltiorrhiza polysaccharides groups(500,250,125 mg/kg),with 12 mice in each group.The treatment group were given salvia miltiorrhiza polysaccharides daily for 7 days.Immunological liver injury in mice was induced by LPS,at 2 hafter administration of LPS,the mouses were killed.Liver pathological changes were observed by HE staining.The activity of serum ALT,AST and the content of TNF-α,INF-γ,IL-10 and IL-6 were determined by ELISA;Using RT-PCR to detect the mRNA expression of TNF-αand INF-γin liver.Compared with the normal group,the activity of ALT,AST,the content of TNF-α,INF-γ,IL-10 and IL-6 in serum,and the mRNA expression of TNF-αand INF-γin liver were increased.Salvia miltiorrhiza polysaccharides(500,250,125 mg/kg)could obviously reduce the activities of ALT,AST,the content of TNF-αand IL-6 in serum and the mRNA expression of TNF-αand INF-γin liver.Salvia miltiorrhiza polysaccharides has a protective effect on immunological liver injury,which is partially via attenuating inflammatory effect.
The aim of the present study was to study the protective effect of salvia miltiorrhiza polysaccharides on immunological liver injury induced by LPS,and to explore a possible mechanism.60 Kunming mouse were randomly divided into 5 groups:the normal group,the model group,the different does of salvia miltiorrhiza polysaccharides groups(500,250,125 mg/kg),with 12 mice in each group.The treatment group were given salvia miltiorrhiza polysaccharides daily for 7 days.Immunological liver injury in mice was induced by LPS,at 2 hafter administration of LPS,the mouses were killed.Liver pathological changes were observed by HE staining.The activity of serum ALT,AST and the content of TNF-α,INF-γ,IL-10 and IL-6 were determined by ELISA;Using RT-PCR to detect the mRNA expression of TNF-αand INF-γin liver.Compared with the normal group,the activity of ALT,AST,the content of TNF-α,INF-γ,IL-10 and IL-6 in serum,and the mRNA expression of TNF-αand INF-γin liver were increased.Salvia miltiorrhiza polysaccharides(500,250,125 mg/kg)could obviously reduce the activities of ALT,AST,the content of TNF-αand IL-6 in serum and the mRNA expression of TNF-αand INF-γin liver.Salvia miltiorrhiza polysaccharides has a protective effect on immunological liver injury,which is partially via attenuating inflammatory effect.
引文
[1]呼晨,孙铭君,姚金水.北冬虫夏草多糖对四氯化碳致小鼠急性肝损伤的影响[J].中国兽医学报,2016,36(3):518-521.
[2]白华,蔡亚娜,胡明,等.芽孢杆菌发酵五味子提取液对肉鸡肝损伤的影响[J].中国兽医学报,2012,32(2):305-308+320.
[3]倪耀娣,乔健,徐建忠,等.紫锥菊、黄芪对法氏囊病雏鸡的肝损伤及脾脏NK细胞活性的影响[J].中国兽医学报,2011,31(4):561-564.
[4]姚莹华,刘强,陈育尧,等.丹参多糖对小鼠急性肝损伤的保护作用[J].中国实验方剂学杂志,2010,16(6):227-230.
[5]张志伟,吴在德.丹参对大鼠肝缺血后肝再生促进作用的初步观察[J].同济医科大学学报,1995,24(4):272-275.
[6]胡义杨,刘平,刘成,等.丹酚酸A抗四氯化碳中毒致大鼠肝损伤和肝纤维化作用[J].中国药理学报,1997,18(5):478-480.
[7]巴翠晶,段雪磊,李得鑫,等.丹参多糖对雏鸡肝损伤的保护作用[J].中国兽医学报,2016,36(10):1737-1740.
[8]SONG Y H,LIU Q,LV Z,et al.Protection of a polysaccharide fromSalvia miltiorrhiza,a Chinese medicinal herb,against immunologicalliver injury in mice[J].Int J Biol Macromol,2008,43(2):170.
[9]徐光尧.肝脏的生理功能[J].安徽医科大学学报,1978(5):18-23.
[10]姜辉,高家荣,刘新平,等.三七总皂苷抗小鼠免疫性肝损伤的研究[J].中药药理与临床,2011,27(4):33-36.
[11]付丹阳,刘雨哲,曲极冰,等.丹参磷脂浸膏对急性四氯化碳肝损伤小鼠肝组织的保护作用[J].中国实验诊断学,2014(11):1741-1743.
[12]YAN Z Q.Regulation of TLR4expression in a tale about tail arteriosclesosis[J].Thromb Vasc Biol,2006,26(4):2582-2584.
[13]SUN R,TIAN Z,KULKARNI S,et al.IL-6prevents T cell-mediated hepatitis via inhibition of NKT cells in CD4+T Cell and STAT3dependent manners[J].J Immunol,2004,172(9):5648-5655.
[14]BLISS S K,BLISS S P,BEITING D P.IL-10Regulates movement of intestinally derived CD4+T cells to the liver[J].J Immunol,2007,178(12):7974-7983.
[2]白华,蔡亚娜,胡明,等.芽孢杆菌发酵五味子提取液对肉鸡肝损伤的影响[J].中国兽医学报,2012,32(2):305-308+320.
[3]倪耀娣,乔健,徐建忠,等.紫锥菊、黄芪对法氏囊病雏鸡的肝损伤及脾脏NK细胞活性的影响[J].中国兽医学报,2011,31(4):561-564.
[4]姚莹华,刘强,陈育尧,等.丹参多糖对小鼠急性肝损伤的保护作用[J].中国实验方剂学杂志,2010,16(6):227-230.
[5]张志伟,吴在德.丹参对大鼠肝缺血后肝再生促进作用的初步观察[J].同济医科大学学报,1995,24(4):272-275.
[6]胡义杨,刘平,刘成,等.丹酚酸A抗四氯化碳中毒致大鼠肝损伤和肝纤维化作用[J].中国药理学报,1997,18(5):478-480.
[7]巴翠晶,段雪磊,李得鑫,等.丹参多糖对雏鸡肝损伤的保护作用[J].中国兽医学报,2016,36(10):1737-1740.
[8]SONG Y H,LIU Q,LV Z,et al.Protection of a polysaccharide fromSalvia miltiorrhiza,a Chinese medicinal herb,against immunologicalliver injury in mice[J].Int J Biol Macromol,2008,43(2):170.
[9]徐光尧.肝脏的生理功能[J].安徽医科大学学报,1978(5):18-23.
[10]姜辉,高家荣,刘新平,等.三七总皂苷抗小鼠免疫性肝损伤的研究[J].中药药理与临床,2011,27(4):33-36.
[11]付丹阳,刘雨哲,曲极冰,等.丹参磷脂浸膏对急性四氯化碳肝损伤小鼠肝组织的保护作用[J].中国实验诊断学,2014(11):1741-1743.
[12]YAN Z Q.Regulation of TLR4expression in a tale about tail arteriosclesosis[J].Thromb Vasc Biol,2006,26(4):2582-2584.
[13]SUN R,TIAN Z,KULKARNI S,et al.IL-6prevents T cell-mediated hepatitis via inhibition of NKT cells in CD4+T Cell and STAT3dependent manners[J].J Immunol,2004,172(9):5648-5655.
[14]BLISS S K,BLISS S P,BEITING D P.IL-10Regulates movement of intestinally derived CD4+T cells to the liver[J].J Immunol,2007,178(12):7974-7983.