Expression of Mindin, a pattern recognition molecule, in a mouse model of acute liver injury induced by carbon tetrachloride
-
摘要:
目的研究Mindin蛋白在CCl4诱导的小鼠急性肝损伤过程中的动态表达情况,并探讨其作用机制。方法雄性C57B1/6小鼠48只,随机抽取40只应用CCl4橄榄油溶液腹腔注射诱导急性肝损伤,分别于造模后6、12、24、48和72 h取小鼠肝脏组织作病理学观察(实验组),余下8只腹腔注射橄榄油作为对照(对照组)。应用Western Blot法检测Mindin蛋白的表达以及变化趋势,实时荧光定量PCR检测肝组织Mindin mRNA的表达。计量资料两组间比较采用t检验。结果 CCl4诱导肝损伤后,小鼠肝脏失去其正常结构,48 h时肝脏病理改变最为显著。肝脏Mindin蛋白在注射CCl4后12~72 h内呈现低表达水平,而其转录水平即Mindin mRNA在给予CCl4后12 h降至最低(实验组vs对照组:0. 183±0. 105 vs 1. 023±0. 247,t=8. 841,P <0. 01),随后逐渐升高,在48 h(2. 548±0. 775 vs 1. 023±0. 247)和72 h(2. 699±0. 995 vs 1. 023±0. 247)时转录水平甚至是正常水平...
Abstract:Objective To investigate the dynamic expression of Mindin protein in mice with acute liver injury induced by carbon tetrachloride ( CCl4) and its mechanism of action. Methods A total of 48 male C57 B1/6 mice were selected and divided into experimental group ( n= 40) and control group ( n = 8) . The mice in the experimental group were given intraperitoneal injection of CCl4 olive oil to induce acute liver injury, and the liver tissue was collected for pathological observation at 6, 12, 24, 48, and 72 hours after modeling. The mice in the control group were given intraperitoneal injection of olive oil. Western blot was used to measure the protein expression of Mindin and its change trend, and quantitative real-time PCR was used to measure the mRNA expression of Mindin. The t-test was used for comparison of continuous data between two groups. Results Abnormal liver structure was observed after liver injury was induced by CCl4, with the most significant pathological change at 48 hours. There was a low protein expression level of Mindin within 12-72 hours after the injection of CCl4. The mRNA expression of Mindin reached the lowest level at 12 hours after CCl4 injection and there was a significant difference between the experimental group and the control group at this time point ( 0. 183 ± 0. 105 vs 1. 023 ± 0. 247, t = 8. 841, P < 0. 01) ; then there was a gradual increase in the mRNA expression of Mindin, and at 48 and 72 hours, the mRNA expression of Mindin in the experimental group was more than 2 times that in the control group ( 48 hours: 2. 548 ± 0. 775 vs 1. 023 ± 0. 247, t = 5. 428, P < 0. 01; 72 hours: 2. 699 ± 0. 995 vs 1. 023± 0. 247, t = 4. 621, P < 0. 01) . Conclusion There is a significant change in Mindin protein during the process of acute liver injury induced by CCl4 in mice, which is characterized by the downregulation of protein expression and the regulation of post-transcriptional mRNA level, suggesting that Mindin may play an important role in the process of acute liver injury induced by CCl4.
-
Key words:
- acute liver injury /
- extracellular matrix proteins /
- mice, inbred strains
-
[1]NAM JS, PARK E, TURCOTTE TJ, et al.Mouse R-spondin2is required for apical ectodermal ridge maintenance in the hindlimb[J].Dev Biol, 2007, 311 (1) :124-135. [2]FEINSTEIN Y, BORRELL V, GARCIA C, et al.F-spondin and mindin:Two structurally and functionally related genes expressed in the hippocampus that promote outgrowth of embryonic hippocampal neurons[J].Development, 1999, 126 (16) :3637-3648. [3]SIMON I, LIU Y, KRALL KL, et al.Evaluation of the novel serum markers B7-H4, Spondin 2, and DcR3 for diagnosis and early detection of ovarian cancer[J].Gynecol Oncol, 2007, 106 (1) :112-118. [4]PARRY R, SCHNEIDER D, HUDSON D, et al.Identification of a novel prostate tumor target, mindin/RG-1, for antibodybased radiotherapy of prostate cancer[J].Cancer Res, 2005, 65 (18) :8397-8405. [5]JIN C, LIN JR, MA L, et al.Elevated spondin-2 expression correlates with progression and prognosis in gastric cancer[J].Oncotarget, 2017, 8 (6) :10416-10424. [6]LUO JH, REN B, KERYANOV S, et al.Transcriptomic and genomic analysis of human hepatocellular carcinomas and hepatoblastomas[J].Hepatology, 2006, 44 (4) :1012-1024. [7]WANG LF, LIU YS, YANG B, et al.The extracellular matrix protein mindin attenuates colon cancer progression by blocking angiogenesis via Egr-1-mediated regulation[J].Oncogene, 2018, 37 (5) :601-615. [8]QIAN X, LI C, PANG B, et al.Spondin-2 (SPON2) , a more prostate-cancer-specific diagnostic biomarker[J].PLo SOne, 2012, 7 (5) :e37225. [9]KAHVECIOGLU S, GUCLU M, USTUNDAG Y, et al.Evaluation of serum Spondin 2 levels in the different stages of Type 2diabetic nephropathy[J].Nephrology (Carlton) , 2015, 20 (10) :721-726. [10]MURAKOSHI M, TANIMOTO M, GOHDA T, et al.Mindin:Anovel marker for podocyte injury in diabetic nephropathy[J].Nephrol Dial Transplant, 2011, 26 (7) :2153-2160. [11]DOGˇAN T, YETIM M, ELIK O, et al.Investigation of mindin levels in hypertensive patients with left ventricular hypertrophy and QRS fragmentation on electrocardiography[J].Acta Cardiol, 2017:1-6. [12]YAN L, WEI X, TANG QZ, et al.Cardiac-specific mindin overexpression attenuates cardiac hypertrophy via blocking AKT/GSK3βand TGF-β1-Smad signalling[J].Cardiovasc Res, 2011, 92 (1) :85-94. [13]GULENG B, LIAN YM, REN JL.Mindin is upregulated during colitis and may activate NF-kappaB in a TLR-9 mediated manner[J].World J Gastroenterol, 2010, 16 (9) :1070-1075. [14]ZHANG C, QIN JJ, GONG FH, et al.Mindin deficiency in macrophages protects against foam cell formation and atherosclerosis by targeting LXR-β[J].Clin Sci (Lond) , 2018, 132 (11) :1199-1213. [15]ZHU LH, HUANG L, ZHANG X, et al.Mindin regulates vascular smooth muscle cell phenotype and prevents neointima formation[J].Clin Sci (Lond) , 2015, 129 (2) :129-145. [16]HE YW, LI H, ZHANG J, et al.The extracellular matrix protein mindin is a pattern-recognition molecule for microbial pathogens[J].Nat Immunol, 2004, 5 (1) :88-97. [17]JIA W, LI H, HE YW.Pattern recognition molecule mindin promotes intranasal clearance of influenza viruses[J].J Immunol, 2008, 180 (9) :6255-6261. [18]LI H, OLIVER T, JIA W, et al.Efficient dendritic cell priming of T lymphocytes depends on the extracellular matrix protein mindin[J].EMBO J, 2006, 25 (17) :4097-4107. [19]SUN P, ZHANG P, WANG PX, et al.Mindin deficiency protects the liver against ischemia/reperfusion injury[J].J Hepatol, 2015, 63 (5) :1198-1211. [20]ZHU LH, WANG A, LUO P, et al.Mindin/Spondin 2 inhibits hepatic steatosis, insulin resistance, and obesity via interaction with peroxisome proliferator-activated receptorαin mice[J].J Hepatol, 2014, 60 (5) :1046-1054. [21]MATZINGER P.The danger model:A renewed sense of self[J].Science, 2002, 296 (5566) :301-305. [22]ZHANG X, DANG SS.Research advances in the mechanism of experimental liver injury[J/CD].Chin J Liver Dis:Electr Version, 2009, 1 (2) :60-62. (in Chinese) 张欣, 党双锁.实验性肝损伤机制研究进展[J/CD].中国肝脏病杂志:电子版, 2009, 1 (2) :60-62. [23]LUO L, ZHANG QF, ZHANG DZ.Advances in the treatment of acute liver failure[J].J Clin Hepatol, 2018, 34 (2) :438-443. (in Chinese) 骆玲, 张琼方, 张大志.急性肝衰竭的治疗进展[J].临床肝胆病杂志, 2018, 34 (2) :438-443. [24]LONG Y, LUO JF, HU MY, et al.Protective effect of ellagic acid on acute liver injury induced by CCl4in mice and its mechanism[J].J Jilin Univ:Med Edit, 2017, 43 (3) :572-576. (in Chinese) 龙毅, 骆静方, 胡敏予, 等.鞣花酸对CCl4诱导小鼠急性肝损伤的保护作用及其机制[J].吉林大学学报:医学版, 2017, 43 (3) :572-576. [25]WANG WJ, DANG SS, ZHAI S, et al.Down-regulated expression of pattern recognition molecule mindin in Huh7 and Huh7.5cells harboring in vitro transcribed full-length hepatitis C virus RNA[J].Chin Hepatol, 2013, 18 (2) :85-87. (in Chinese) 王文俊, 党双锁, 翟嵩, 等.模式识别分子mindin在体外HCV全基因组转染的Huh-7和Huh-7.5细胞系中表达下调[J].肝脏, 2013, 18 (2) :85-87.
计量
- 文章访问数: 1689
- HTML全文浏览量: 10
- PDF下载量: 273
- 被引次数: 0