丹参多糖缓解免疫性肝损伤的机制
|
摘要
为探究丹参多糖缓解肝损伤的机制,本研究将50只昆明小鼠随机分为正常对照组(A),LPS模型组(B),丹参多糖各剂量保护组500(C),250(D),125 mg/kg(E),每组10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,处死小鼠摘取肝组织。RT-PCR检测肝组织中CXCL-10和iNOS mRNA的表达;Westiern blot测定肝组织中NF-κB,CXCL-10和iNOS蛋白的表达。结果显示,与空白组相比,LPS能显著性提高肝组织中CXCL-10和iNOS mRNA及其蛋白的表达量(P<0.01),可显著性促进NF-κB中P65和IκBα的磷酸化水平(P<0.01)。而丹参多糖提前处理可以并显著性降低肝组织中CXCL-10和iNOS mRNA及其蛋白的表达量(P<0.01或P<0.05),并能显著抑制P65和IκBα在体内外的磷酸化水平的表达(P<0.01或P<0.05)。结果表明,丹参多糖可以通过抑制炎症通路NF-κB和趋化因子CXCL-10的活性来降低炎症因子的释放达到抑制炎症反应的作用,进而缓解免疫性肝损伤。
The aim of the present study was to explore the mechanism of Salvia miltiorrhiza polysaccharide(SMP)relieving immune liver injury.Fifty Kunming mice were randomly divided into 5 groups,the control group(A),the LPS model group(B),the different doses of SMP groups(group C 500 mg/kg,group D 250 mg/kg,group E 125 mg/kg),with 10 mice of each group.Mice of the groups C,D,E were given Salvia miltiorrhiza polysaccharides daily for 7 days consecutively.Immunological liver injury in mice was induced by LPS.At two hours after administration of LPS,the mouses were killed to sample liver.The mRNA expressions of CXCL-10 and iNOS were detected by using RT-PCR.The expressions of NF-κB,CXCL-10 and iNOS were measured by Western blot.Compared with the control group,the mRNA and protein expressions of CXCL-10 and iNOS were upregulated.The expressions of p-p65 and p-IκBαalso were elevated.Salvia miltiorrhiza polysaccharides(at 500,250,125 mg/kg)significantly reduce the mRNA and protein expressions of CXCL-10 and iNOS.The expressions of p-p65 and p-IκBαwere downregulated in the SMP group compared with the model group.The results of this study indicate Salvia miltiorrhiza polysaccharides to relese inflammamtion partially via attenuating chemokines and adjusting the NF-κB pathway.
The aim of the present study was to explore the mechanism of Salvia miltiorrhiza polysaccharide(SMP)relieving immune liver injury.Fifty Kunming mice were randomly divided into 5 groups,the control group(A),the LPS model group(B),the different doses of SMP groups(group C 500 mg/kg,group D 250 mg/kg,group E 125 mg/kg),with 10 mice of each group.Mice of the groups C,D,E were given Salvia miltiorrhiza polysaccharides daily for 7 days consecutively.Immunological liver injury in mice was induced by LPS.At two hours after administration of LPS,the mouses were killed to sample liver.The mRNA expressions of CXCL-10 and iNOS were detected by using RT-PCR.The expressions of NF-κB,CXCL-10 and iNOS were measured by Western blot.Compared with the control group,the mRNA and protein expressions of CXCL-10 and iNOS were upregulated.The expressions of p-p65 and p-IκBαalso were elevated.Salvia miltiorrhiza polysaccharides(at 500,250,125 mg/kg)significantly reduce the mRNA and protein expressions of CXCL-10 and iNOS.The expressions of p-p65 and p-IκBαwere downregulated in the SMP group compared with the model group.The results of this study indicate Salvia miltiorrhiza polysaccharides to relese inflammamtion partially via attenuating chemokines and adjusting the NF-κB pathway.
引文
[1]李峰,朱洁平,王艳梅,等.天麻多糖对小鼠免疫性肝损伤的保护作用[J].中药药理与临床,2015(1):111-113.
[2]王忠利,王洪新.黄芪多糖对免疫性肝损伤大鼠的保护作用研究[J].中药药理与临床,2013(2):77-80.
[3]宋雨鸿,刘强,徐舒,等.丹参多糖对小鼠免疫性肝损伤的保护作用[J].中南药学,2013(5):345-348.
[4]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(10):1737-1740.
[5]胡宝吉.红景天苷预处理对刀豆蛋白A诱导小鼠肝损伤的作用及机制探讨[D].上海:第二军医大学,2014.
[6]RECKNAGEL P,GONNERT F A,HALILBASIC E,et al.Mechanisms and functional consequences of liver failure substantially differ between endotoxaemia and faecal peritonitis in rats[J].Liver Int,2013,33(2):283-293.
[7]LI Y,WANG X,WEI Z,et al.Pretreatment with wortmannin alleviates lipopolysaccharide/d-galactosamine-induced acute liver injury[J].Biochem Biophys Res Commun,2014,455(3/4):234-240.
[8]YAN Z Q.Regulation of TLR4expression is a tale about tail[J].Arterioscler Thromb Vasc Biol,2006,26(12):2582-2584.
[9]CAO W,ZHANG W,LIU J,et al.Paeoniflorin improves survival in LPS-challenged mice through the suppression of TNF-alpha and IL-1beta release and augmentation of IL-10production[J].Int Immunopharmacol,2011,11(2):172-178.
[10]KANG S M,MORE S V,PARK J Y,et al.A novel synthetic HTB derivative,BECT inhibits lipopolysaccharide-mediated inflammatory response by suppressing the p38 MAPK/JNK and NF-kappaB activation pathways[J].Pharmacol Rep,2014,66(3):471-479.
[11]KIM I T,RYU S,SHIN J S,et al.Euscaphic acid isolated from roots of Rosa rugosa inhibits LPS-induced inflammatory responses via TLR4-mediated NF-kappaB inactivation in RAW 264.7macrophages[J].J Cell Biochem,2012,113(6):1936-1946.
[12]WELSH P,GRASSIA G,BOTHA S,et al.Targeting inflammation to reduce cardiovascular disease risk:a realistic clinical prospect?[J].Br J Pharmacol,2017,174(22):3898-3913.
[13]ZHOU L,ZHU D Y.Neuronal nitric oxide synthase:structure,subcellular localization,regulation,and clinical implications[J].Nitric Oxide,2009,20(4):223-230.
[14]MEDZHITOV R.Toll-like receptors and innate immunity[J].Nat Rev Immunol,2001,1(2):135-145.
[15]TU C T,HAN B,LIU H C,et al.Curcumin protects mice against concanavalin A-induced hepatitis by inhibiting intrahepatic intercellular adhesion molecule-1(ICAM-1)and CXCL10expression[J].Mol Cell Biochem,2011,358(1/2):53-60.
[16]CAO L,ZOU Y,ZHU J,et al.Ginsenoside Rg1attenuates concanavalin A-induced hepatitis in mice through inhibition of cytokine secretion and lymphocyte infiltration[J].Mol Cell Biochem,2013,380(1/2):203-210.
[17]MAGHAZACHI A A.Role of chemokines in the biology of natural killer cells[J].Curr Top Microbiol Immunol,2010,341:37-58.
[18]LEE E Y,LEE Z H,SONG Y W.CXCL10and autoimmune diseases[J].Autoimmun Rev,2009,8(5):379-383.
[19]ANTONELLI A,FERRARI S M,GIUGGIOLI D,et al.Chemokine(C-X-C motif)ligand(CXCL)10in autoimmune diseases[J].Autoimmun Rev,2014,13(3):272-280.
[20]KAKIMI K,LANE T E,WIELAND S,et al.Blocking chemokine responsive to gamma-2/interferon(IFN)-gamma inducible protein and monokine induced by IFN-gamma activity in vivo reduces the pathogenetic but not the antiviral potential of hepatitis B virus-specific cytotoxic T lymphocytes[J].J Exp Med,2001,194(12):1755-1766.
[21]BIECHE I,ASSELAH T,LAURENDEAU I,et al.Molecular profiling of early stage liver fibrosis in patients with chronic hepatitis C virus infection[J].Virology,2005,332(1):130-144.
[2]王忠利,王洪新.黄芪多糖对免疫性肝损伤大鼠的保护作用研究[J].中药药理与临床,2013(2):77-80.
[3]宋雨鸿,刘强,徐舒,等.丹参多糖对小鼠免疫性肝损伤的保护作用[J].中南药学,2013(5):345-348.
[4]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(10):1737-1740.
[5]胡宝吉.红景天苷预处理对刀豆蛋白A诱导小鼠肝损伤的作用及机制探讨[D].上海:第二军医大学,2014.
[6]RECKNAGEL P,GONNERT F A,HALILBASIC E,et al.Mechanisms and functional consequences of liver failure substantially differ between endotoxaemia and faecal peritonitis in rats[J].Liver Int,2013,33(2):283-293.
[7]LI Y,WANG X,WEI Z,et al.Pretreatment with wortmannin alleviates lipopolysaccharide/d-galactosamine-induced acute liver injury[J].Biochem Biophys Res Commun,2014,455(3/4):234-240.
[8]YAN Z Q.Regulation of TLR4expression is a tale about tail[J].Arterioscler Thromb Vasc Biol,2006,26(12):2582-2584.
[9]CAO W,ZHANG W,LIU J,et al.Paeoniflorin improves survival in LPS-challenged mice through the suppression of TNF-alpha and IL-1beta release and augmentation of IL-10production[J].Int Immunopharmacol,2011,11(2):172-178.
[10]KANG S M,MORE S V,PARK J Y,et al.A novel synthetic HTB derivative,BECT inhibits lipopolysaccharide-mediated inflammatory response by suppressing the p38 MAPK/JNK and NF-kappaB activation pathways[J].Pharmacol Rep,2014,66(3):471-479.
[11]KIM I T,RYU S,SHIN J S,et al.Euscaphic acid isolated from roots of Rosa rugosa inhibits LPS-induced inflammatory responses via TLR4-mediated NF-kappaB inactivation in RAW 264.7macrophages[J].J Cell Biochem,2012,113(6):1936-1946.
[12]WELSH P,GRASSIA G,BOTHA S,et al.Targeting inflammation to reduce cardiovascular disease risk:a realistic clinical prospect?[J].Br J Pharmacol,2017,174(22):3898-3913.
[13]ZHOU L,ZHU D Y.Neuronal nitric oxide synthase:structure,subcellular localization,regulation,and clinical implications[J].Nitric Oxide,2009,20(4):223-230.
[14]MEDZHITOV R.Toll-like receptors and innate immunity[J].Nat Rev Immunol,2001,1(2):135-145.
[15]TU C T,HAN B,LIU H C,et al.Curcumin protects mice against concanavalin A-induced hepatitis by inhibiting intrahepatic intercellular adhesion molecule-1(ICAM-1)and CXCL10expression[J].Mol Cell Biochem,2011,358(1/2):53-60.
[16]CAO L,ZOU Y,ZHU J,et al.Ginsenoside Rg1attenuates concanavalin A-induced hepatitis in mice through inhibition of cytokine secretion and lymphocyte infiltration[J].Mol Cell Biochem,2013,380(1/2):203-210.
[17]MAGHAZACHI A A.Role of chemokines in the biology of natural killer cells[J].Curr Top Microbiol Immunol,2010,341:37-58.
[18]LEE E Y,LEE Z H,SONG Y W.CXCL10and autoimmune diseases[J].Autoimmun Rev,2009,8(5):379-383.
[19]ANTONELLI A,FERRARI S M,GIUGGIOLI D,et al.Chemokine(C-X-C motif)ligand(CXCL)10in autoimmune diseases[J].Autoimmun Rev,2014,13(3):272-280.
[20]KAKIMI K,LANE T E,WIELAND S,et al.Blocking chemokine responsive to gamma-2/interferon(IFN)-gamma inducible protein and monokine induced by IFN-gamma activity in vivo reduces the pathogenetic but not the antiviral potential of hepatitis B virus-specific cytotoxic T lymphocytes[J].J Exp Med,2001,194(12):1755-1766.
[21]BIECHE I,ASSELAH T,LAURENDEAU I,et al.Molecular profiling of early stage liver fibrosis in patients with chronic hepatitis C virus infection[J].Virology,2005,332(1):130-144.