Pristane诱导BALB/c小鼠系统性红斑狼疮模型的建立
Experimental study of pristane-induced BALB/c mice systemic lupus erythematosus model
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摘要: 目的:建立Pristane诱导的BALB/c小鼠系统性红斑狼疮(SLE)模型。方法:雌性BALB/c小鼠随机分成2组,模型组单次腹腔注射0.5 ml Pristane,对照组注射等量的0.9%氯化钠注射液,注射前及注射后每月ELISA法检测血清抗dsDNA抗体和抗Sm/RNP抗体含量,Albustix试纸法检测尿蛋白含量,每月定期观察小鼠症状和体征。6个月后处死全部小鼠后解剖,肉眼观察腹腔脏器组织病变,并取病变组织及肾脏做病理学检查,观察其组织病理变化(HE染色法)及肾脏免疫复合物(IC)沉积情况(直接荧光染色法)。结果:模型组小鼠造模2个月后,血清抗Sm/RNP抗体和抗dsDNA抗体开始出现,并逐月增高,与同期对照组小鼠比较,3～6个月时血清抗Sm/RNP抗体和4～6个月时血清抗dsDNA抗体均明显增高(P0.01),且血清抗Sm/RNP抗体的增高较抗dsDNA抗体更为显著;尿蛋白(+)于造模后3个月时开始出现,6个月时显著高于对照组小鼠(P0.01);3个月时模型组小鼠开始出现关节病变,6个月时其阳性率达55%,明显高于对照组小鼠(P=0.004)。6个月后处死动物,解剖发现模型组大多数小鼠腹腔可见多少不等的脂肪肉芽肿结节;肾脏病理检查50%的模型组小鼠出现不同程度肾小球肾炎病变伴毛细血管壁大量IC沉积。对照组除1只小鼠在造模6个月时出现轻度蛋白尿(+)外,未见其他病变。结论:Pristane能成功诱发BALB/c小鼠SLE,且建立的SLE模型稳定可靠。Abstract: Objective: To establish a mice systemic lupus erythematosus(SLE) model induced by Pristane.Methods: Female BALB/ c mice were divided into two groups randomly.The mice in model group were received a single intraperitoneal injection of 0.5 ml Pristane,while the mice in control group were received 0.9% sodium chloride injection with same volume.Before and after injection,anti-Sm / RNP antibody and anti-dsDNA antibody in serum were detected by ELISA,proteinuria content was quantified by Albustix dipstick test,and the changes of signs and symptoms in these mice were monitored monthly.Six months after injection,all mice were killed and lesions in abdominal organs were checked by anatomy.All injured organs were observed and kidneys were inspected by pathology.Histopathologic changes of injured organs were determined by HE staining method,and glomerular IgG deposition in kidney was detected by direct immunofluorescence staining method.Results: Serum anti-Sm / RNP antibody and anti-dsDNA antibody were detected two months after Pristane injection,and increased monthly.Compared with the control mice in the corresponding period,serum anti-Sm / RNP antibody and anti-dsDNA antibody in the model mice were increased obviously at 3-6 months after injection(P 0.01) and at 4-6 months after injection respectively(P 0.01).The increase of serum anti-Sm / RNP antibody was significantly higher than that of serum anti-dsDNA antibody.Urine protein( +) was detected since 3 months after Pristane injection,and at 6 months,urine protein content was significantly higher than that in the control mice(P 0.01).The symptom of arthritis in legs was shown at the third months after Pristane injection,and the positive incidence was 55% after 6 months,apparently higher than that in the control mice(P = 0.004).After the mice were killed at the sixth months,abdominal lipogranulomas more or less were found in most of the model mice.Kidney pathological examination was shown in over 50% Pristane-treated mice,glomerulonephritis more or less were observed and obvious immune complex deposition was observed in the wall of capillaries.Whereas,with an exception of one mouse developing proteinuria at 6th month after injection,none of the control mice displayed any pathological changes.Conclusions: Pristane can successfully induce mice SLE in BALB / c by intraperitoneal injection and is steady and reliable.
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Key words:
- lupus erythematosus,systemic /
- Pristane /
- experimental model /
- mice
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[1] Gesikowska K,Kondera-Anasz Z,Mielczarek-Palacz A,et al.Systemic lupus erythematosus-still current clinical and diagnostic problem[J].Pol Merkur Lekarski,2012, 32( 188):111-115. [2] Belot A,Cimaz R.Monogenic forms of systemic lupus erythematosus: new insights into SLE pathogenesis[J].Pediatr Rheumatol Online J,2012,10( 1):21. [3] Perry D,Sang A,Yin Y, et al.Murine models of systemic lupus erythematosus[J].J Biomed Biotechnol,2011: 271694. [4] Reeves WH,Lee PY,Weinstein JS, et al.Induction of autoimmunity by pristane and other naturally occurring hydrocarbons[J].Trends Immunol,2009,30( 9):455-464. [5] Satoh M,Richards HB, Shaheen VM, et al.Widespread susceptibility among inbred mouse strains to the induction of lupus autoantibodies by pristane[J].Clin Exp Immunol, 2000,121( 2):399-405. [6] Szczepańiska M,Muszewska E,Szprynger K, et al.Systemic lupus erythematosus and pregnancy[J].Wiad Lek,2008,61 ( 4 /6 ):161-165. [7] Kim HA,Chung JW,Park HJ,et al.An antinuclear antibodynegative patient with lupus nephritis[J].Korean J Intern Med,2009,24(1):76-79. [8] Richards HB,Satoh M,Shaw M, et al.Interleukin 6 dependence of anti-DNA antibody production: evidence for two pathways of autoantibody formation in pristane-induced lupus[J].J Exp Med,1998,188( 5):985-990. [9] Bultink IE,Vis M, van der Horst-Bruinsma IE, et al.Inflammatory rheumatic disorders and bone[J].Curr Rheumatol Rep,2012,14(3):224-230. [10] Hochberg MC.Updating the American College of Rheumatology revised criteria for the classification of systemic lupus erythematosus[J].Arthritis Rheum,1997,40( 9):1725. [11] Lee PY,Weinstein JS,Nacionales DC, et al.A novel type Ⅰ IFNproducing cell subset in murine lupus[J].J Immunol,2008,180(7):5101-5108. [12] Calvani N,Caricchio R,Tucci M, et al.Induction of apoptosis by the hydrocarbon oil pristane: implications for pristane-induced lupus[J].J Immunol,2005,175( 7):4777-4782. [13] Herman S,Kny A,Schorn C,et al.Cell death and cytokine production induced by autoimmunogenic hydrocarbon oils[J].Autoimmunity,2012, 42(8):602-611. [14] Janz S,Shacter E.A new method for delivering alkanes to mammalian cells: preparation and preliminary characterization of an inclusion complex between beta-cyclodextrin and pristane( 2,6,10,14-tetramethylpentadecane) [J].Toxicology,1991,69( 3):301-315. [15] Mizutani A,Shaheen VM,Yoshida H,et al.Pristane-induced autoimmunity in germ-free mice [J].Clin Immunol,2005,114( 2):110-118. [16] Janeway CA Jr,Goodnow CC,Medzhitov R.Danger-pathogen on the premises Immunological tolerance [J].Curr Biol,1996,6( 5):519-522. [17] Matzinger P.Tolerance,danger,and the extended family[J].Annu Rev Immunol,1994, 12: 991-1045. -
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