文章摘要
蔡青云,杨留生,张贺川.Toll样受体 2、Toll样受体 4 mRNA和蛋白在急性出血坏死性胰腺炎肝损伤中的表达及意义[J].安徽医药,2022,26(1):82-86.
Toll样受体 2、Toll样受体 4 mRNA和蛋白在急性出血坏死性胰腺炎肝损伤中的表达及意义
Expression and significance of Toll-like receptor 2, Toll-like receptor 4 mRNA and protein in hepatic injury induced by AHNP
  
DOI:10.3969/j.issn.1009-6469.2022.01.019
中文关键词: 胰腺炎,急性坏死性  肝功能不全  丙氨酸转氨酶  Toll样受体 2  Toll样受体 4  信使核糖核酸  蛋白  大鼠, Sprague-Dawley 淀粉,
英文关键词: Pancreatitis,acute necrotizing  Hepatic insufficiency  Alanine transaminase  Toll-like receptor 2  Toll-like recep-tor 4  mRNA  Protein  Rats,sprague-dawley
基金项目:
作者单位
蔡青云 驻马店市中心医院急诊科河南驻马店 463000 
杨留生 驻马店市中心医院急诊科河南驻马店 463000 
张贺川 驻马店市中心医院急诊科河南驻马店 463000 
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中文摘要:
      目的探讨 Toll样受体 2(TLR2)、 TLR4信使核糖核酸( mRNA)和蛋白在急性出血坏死性胰腺炎( AHNP)肝损伤中的表达及意义。方法 2019年 6月至 12月选取健康 SD大鼠(郑州大学实验动物中心) 60只为研究对象,采用随机数字表法分为模型组和对照组,每组 30只,同时模型组和对照组再随机分为 3h组、 6h组和 12 h组各 10只。模型组采用牛磺胆酸钠建立 AHNP模型,对照组注射生理盐水,检测各组血清丙氨酸氨基转移酶( ALT)、天门冬氨酸氨基转移酶( AST)和淀粉酶, qRT-PCR和 Western blotting检测肝脏组织 TLR2、TLR4 mRNA和蛋白表达, HE染色观察肝脏组织。结果模型组造模后 3h组、 6h组和 12 h组血清 ALT[( 170.02±32.21)U/L、(310.01±80.02)U/L、(720.11±112.28)U/L]、 AST[( 450.13±100.21)U/L、(980.12±132.21)U/L、(1 100.32±145.03)U/L]和淀粉酶[( 7 099.27±1 209.02)U/L、(11 010.19±1 200.01)U/L、(17 003.21±2 007.82)U/L]肝脏组织 TLR2和 TLR4 mRNA及蛋白相对表达量[ TLR2 mRNA(0.962±0.201)、(1.201±0.223)、(1.501±0.212), TLR2蛋白( 0.71,3±0.091)、(1.102±0.100)、(1.343±0.121); TLR4 mRNA(1.701±0.332)、(2.011±0.350)、(2.562±0.313),TLR4蛋白( 1.210±0.170)、(1.446±0.182)、(1.788±0.190)]均逐渐增高( P<0.001),且均明显高于对照组[ ALT(84.40±14.92)U/L、(85.50± 15.20)U/L、(84.19±14.49)U/L;AST(170.02±20.01)U/L、(168.82±21.14)U/L、(171.14±20.11)U/L;淀粉酶( 1 102.00±183.29)U/ L、(1 154.49±190.04)U/L、(1 160.03±186.27)U/L;TLR2 mRNA(0.030±0.010)、(0.031±0.011)、(0.029±0.009)TLR2蛋白( 0.021± 0.008)、(0.020±0.009)、(0.022±0.010); TLR4 mRNA(0.882±0.181)、(0.880±0.170)、(0.883±0.179)TLR4(0.961±0.192)、蛋白,(0.967±0.181)、(0.966±0.180)](P<0.001)。肝组织 TLR2、TLR4 mRNA表达与血清 ALT、AST和酶呈正相关( P<0.05), TLR2、TLR4蛋白表达与血清 ALT、AST和淀粉酶呈正相关(P<0.05);模型组造模 3h后可见肝细胞气球样变性, 6h后炎性细胞浸润汇管区,小叶周边细胞发生嗜酸性变性, 12 h后出现大片状出血坏死及灶性坏死。结论 TLR2、TLR4 mRNA和蛋白在 AHNP肝损伤组织中的表达升高,可能在肝脏损伤过程中有重要作用。
英文摘要:
      Objective To investigate the expression and significance of Toll-like receptor 2 (TLR2), Toll-like receptor 4 (TLR4) mRNA and protein in liver injury of acute hemorrhagic necrotizing pancreatitis (AHNP).Methods Sixty healthy SD rats in Experimen-tal Animal Center of Zhengzhou University from June to December 2019 were selected as subjects. The rats were assigned into modelgroup and control group by random number table method, with 30 rats in each group. Meanwhile, the model group and control groupwere randomly divided into 3 h group, 6 h group and 12 h group with 10 rats in each group. The model group was established with SO-DIUM taurocholate to establish AHNP model, while the control group was injected with normal saline. Serum alanine aminotransferase(ALT), aspartate aminotransferase (AST) and amylase of the different groups were detected. The mRNA and protein expressions ofTLR2 and TLR4 in liver tissues were detected by QRT-PCR and Western blotting, and the liver tissues were observed by HE staining. Results Serum ALT [(170.02±32.21) U/L, (310.01±80.02) U/L, (720.11±112.28) U/L], AST [(450.13±100.21) U/L, (980.12±132.21)U/L, (1 100.32±145.0) U/L] and amylase [(7 099.27±1 209.02) U/L, (11 010.19±1 200.01) U/L, (17 003.21±2 007.82) U/L], TLR2 andTLR4 mRNA and protein relative expression levels in liver tissues [TLR2 mRNA (0.962±0.201), (1.201±0.223), (1.501±0.212), TLR2protein (0.713±0.091), (1.102±0.100), (1.343±0.121); TLR4 mRNA (1.701±0.332), (2.011±0.350), (2.562±0.313), TLR4 protein(1.210±0.170), (1.446±0.182), (1.788±0.190)] of the model group 3 h, 6 h and 12 h after modeling were increased gradually (P<0.001).And they were significantly higher than the control group [ALT (84.40±14.92) U/L, (85.50±15.20) U/L, (84.19±14.49) U/L; AST (170.02±20.01) U/L, (168.82±21.14) U/L, (171.14±20.11) U/L; Amylase (1 102.00±183.29) U/L, (1 154.49±190.04) U/L, (1 160.03±186.27) U/L; TLR2 mRNA (0.030±0.010), (0.031±0.011), (0.029±0.009), TLR2 protein (0.021±0.008), (0.020±0.009), (0.022±0.010);TLR4 mRNA (0.882±0.181), (0.880±0.170), (0.883±0.179), TLR4 protein (0.961±0.192), (0.967±0.181), (0.966±0.180)] (P<0.001). The mRNA expressions of TLR2 and TLR4 in liver tissue were positively correlated with ALT, AST and amylase in serum (P<0.05), and the protein expressions of TLR2 and TLR4 were positively correlated with ALT, AST and amylase in serum (P<0.05). In the modelgroup, balloonlike degeneration of liver cells was observed 3 h after modeling, inflammatory cells infiltrated into the sink area 6 h laterand eosinophilic degeneration occurred in the cells around lobules, and large patellar bleeding necrosis and focal necrosis occurred 12h later.Conclusion The expression of TLR2 and TLR4 mRNA and protein in liver injury tissue induced by AHNP are increase, whichmay play an important role in the process of liver injury.
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