赛培新增产品文献：大鼠TLＲ4、TNF-α、NO ELISA试剂盒

发布时间：2022-04-08 11:56:25

毛蕊异黄酮调控 TLＲ4 /p38MAPK 通路减轻脓毒症 大鼠心肌损伤的机制研究 

刘杰 李萍 高彦霞 吴瑶 项目来源: 陕西省科学技术计划项目( 编号: 2017SF-261) 作者单位: 710004 西安市，西安交通大学第二附属医院急诊科 通讯作者: 李萍，710004 西安市，西安交通大学第二附属医院急 诊科; E-mail: iephpu@ 163． com 【摘要】 目的 观察毛蕊异黄酮减轻脓毒症大鼠心肌损伤的作用，并探讨其是否能够调控 Toll 样受体 4 ( TLＲ4) /丝裂原活化蛋白激酶( p38MAPK) 通路发挥此作用及具体的机制。方法 54 只成年 SD 大鼠随机分为 6 组， 除正常对照组( NC 组) 外其余均建立脓毒症模型。成功后阳性对照组( PC 组) 予以乌司他丁100 kU/kg 溶于1 ml/100 g 0． 9% 氯化钠溶液中经腹腔注射，低、中、高浓度组( A、B、C 组) 分别予以 5、10、20 mg /kg 毛蕊异黄酮溶于 1 ml /100 g 0． 9% 氯化钠溶液中经腹腔注射; 模型对照组( MC 组) 和 NC 组均予以 1 ml /100 g 0． 9% 氯化钠溶液经腹腔注射。 1 次/d，干预 1 周。干预后均处死，对比干预后心肌组织 TLＲ4、肿瘤坏死因子-α( TNF-α) 、一氧化氮( NO) 含量; 苏木 素-伊红染色法( HE) 观察心肌组织病理情况; 脱氧核糖核苷酸末端转移酶介导的缺口末端标记法( TUNEL) 检测心肌 组织细胞凋亡指数( AI) ; 实时聚合酶链反应( ＲT-PCＲ) 检测心肌组织 TLＲ4、p38MAPK mＲNA 表达; 蛋白质免疫印迹法 ( WB) 检测心肌组织 TLＲ4、p38MAPK 蛋白表达和磷酸化 p38MAPK( p-p38MAPK) 水平。结果 MC 组心肌组织 TLＲ4、 TNF-α、NO 含量，心肌细胞 AI 均较 NC 组升高( P ＜ 0． 01) ，PC 组和 3 浓度组均较 MC 组下降( P ＜ 0． 01) ，PC 组、B 组、C 组均较 A 组下降( P ＜ 0． 01) ，C 组均较 PC 组、B 组下降( P ＜ 0． 01) ; MC 组心肌纤维和细胞均呈严重病理改变，PC 组和 3 个浓度组病理变化均显著减轻，且 C 组与 NC 组最为接近; MC 组心肌组织 TLＲ4 mＲNA 及蛋白，p-p38MAPK 水平均 高于 NC 组( P ＜ 0． 01) ，PC 组和 3 个浓度组均低于 MC 组( P ＜ 0． 01) ，PC 组、B 组、C 组均较 A 组下降( P ＜ 0． 01) ，C 组 均较 PC 组和 B 组下降( P ＜ 0． 01) 。结论 对脓毒症大鼠腹腔注射毛蕊异黄酮可控制心肌组织炎性反应，减轻病理改 变，抑制心肌细胞凋亡，且其作用呈浓度依赖性，推测与抑制 TLＲ4 mＲNA 与蛋白表达，控制 p-p38MAPK 有关。 【关键词】 毛蕊异黄酮; Toll 样受体 4; 丝裂原活化蛋白激酶; 脓毒症; 心肌损伤; 炎性反应 【中图分类号】 Ｒ 631 【文献标识码】 A 【文章编号】 1002 － 7386( 2021) 07 － 0965 － 06

Experimental study on the effects of Calycosin on myocardial injury in rats with sepsis by regulating TLＲ4 / p38MAPK pathway LIU Jie，LI Ping，GAO Yanxia，et al． Emergency Department，The Second Affiliated Hospital of Xi’an Jiaotong University，Shaaxi，Xi’an 710004，China 【Abstract】 Objective To investigate the effects of Calycosin on myocardial injury in rats with sepsis，and to explore whether it can regulate Toll-like receptor 4 ( TLＲ4) /mitogen-activated protein kinase ( p38MAPK) pathway and its action mechanism． Methods Fifty-four adult SD rats were randomly divided into six groups． The rat models with sepsis were established except for those in normal control group ( NC group) ． After the rat models were successfully established，the rats in positive control group ( PC group) were injected intraperitoneally with ulinastatin 100kU/kg dissolved in 1ml /100g saline． The rats in low，medium and high dose groups ( group A，B and C) were injected intraperitoneally with 5mg，10mg and 20mg /kg of Calycosin dissolved in 1ml /100g saline，respectively． The rats in model control group ( MC group) and NC group were intraperitoneally injected with 1ml /100g saline，once a day for a week． After intervention，all the rats were sacrificed，and the levels of TLＲ4，TNF-α and NO in myocardial tissue were detected． Hematoxylin-eosin staining ( HE) was used to observe the pathological changes of myocardium． Apoptotic index ( AI) of cardiac myocytes was measured by deoxyribonucleotide terminal transferase mediated nick end labeling ( TUNEL) ． Ｒeal-time polymerase chain reaction ( ＲT-PCＲ) was used to detect the expression levels of TLＲ4 and p38MAPK in myocardium． Moreover the expression levels of TLＲ4，p38MAPK and phosphorylated p38MAPK ( p-p38MAPK) in myocardium were detected by Western Blot． Ｒesults The levels of TLＲ4，TNF- α，NO and AI in myocardial tissues in MC group were significantly higher than those in NC group ( P ＜ 0． 01) ，however，which in PC group and group A，B，C were significantly lower than those in MC group ( P ＜ 0． 01) ，moreover，which in PC group， group B and group C were significantly lower than those in group A ( P ＜ 0． 01) ，and which in group C were significantly lower than those in PC group and group B ( P ＜ 0． 01) ． The pathological changes of myocardial fibers and cells in MC group were serious，however，which in PC group and group A，B，C were significantly alleviated，which in group C were the closest to those in NC group． The levels of TLＲ4 gene and protein，p- p38MAPK in myocardial tissue in MC group were significantly higher than those in NC group ( P ＜ 0． 01) ，which in PC group and group A，B，C were significantly lower than those in MC group ( P ＜ 0． 01) ，however，which in PC group，group B and 965 河北医药 2021 年 4 月 第 43 卷 第 7 期 Hebei Medical Journal，2021，Vol 43 Apr No. 7group C were significantly lower than those in group A ( P ＜ 0． 01) ，which in group C were significantly lower than those in PC group and group B ( P ＜ 0． 01) ． Conclusion The intraperitoneal injection of Calycosin for the rats with sepsis can control myocardial inflammation reaction，alleviate pathological changes and inhibit cardiomyocyte apoptosis in a concentration- dependent manner，and its action mechanism may be related to the inhibition of TLＲ4 gene and protein expression and the control of p-p38MAPK pathway． 【Key words 】 Calycosin; Toll-like receptor 4; mitogen-activated protein kinase; sepsis; myocardial injury; inflammatory reaction

毛蕊异黄酮调控TLR4_p...毒症大鼠心肌损伤的机制研究_刘杰.pdf