The protective effect of L-Shikonin on LPS/D-GalN-induced acute liver injury in mice via inhibiting NF-κB inflammatory signaling pathway北大核心CSCD

Aim To investigate the anti-inflammatory effect of L-Shikonin ( SK ) on lipopolysaccharide ( LPS)-induced RAW 264. 7 macrophages in vitro and its protective effect on LPS/D-GalN-induced acute liver injury. Methods The mouse model of acute liver in¬jury was established in vivo experiments by LPS/D- GalN. The survival rate of the mice and the changes of liver and spleen indices in each group were examined. The levels of AST, ALT and AKP in serum and NO, superoxide dismutase ( SOD ) and malondialdehyde (MDA) in liver tissue homogenate were measured, and the histopathological sections of the liver of each group were observed by H&E staining. M I T colorimet- ric assay was used for cell viability in vitro experi¬ments, Griess method for the detection of NO content, RT-PCR assay and Western blot assay for examining the effect of levulinic acid on the expression levels of mRNA and related pathway proteins of pro-inflammato¬ry factors in LPS-induced RAW264. 7 cells. Results The results of in vivo experiments showed that L-SK significantly improved the liver and spleen indices, de¬creased AST, ALT and AKP levels in serum, de¬creased NO and MDA in liver homogenate, and in¬creased SOD activity in mice with acute liver injury. The results of in vitro experiments showed that L-SK significantly inhibited the mRNA expression of INOS, COX2, I FN-(3 and pro-inflammatory factors 1L-6, TNF-a and IL-10 in LPS-induced RAW264. 7 cells, and significantly inhibited the protein expression of IN¬OS, COX2 and the NF-kB signaling pathway. Conclu¬sions L-SK has good anti-inflammatory effects in LPS-induced inflammation in RAW 264. 7 cells in vitro. Il inhibits the protein expression of phosphorylated P65 and IKKaαβ in the NF-kB signaling pathway, thereby suppressing the anti-inflammatory effects in vitro and L- Shikonin has protective effects against acute liver injury in mice. © 2023 Publication Centre of Anhui Medical University. All rights reserved.     

三叶青黄酮经p38MAPK和NF-κB途径抑制老年小鼠急性肺损伤北大核心CSCD

目的探讨三叶青总黄酮(RTFs)对老年小鼠急性肺损伤(ALI)的保护作用及可能机制。方法采用老年C57BL/6J小鼠支气管滴注脂多糖(LPS)诱导ALI模型,三叶青黄酮灌胃给药3 d。取支气管肺泡灌洗液(BALF),瑞氏-吉姆萨染色计算白细胞数,ELISA检测炎症因子水平;ELISA和Western blot分析肺组织MAPKs、NF-κB蛋白表达,Trans AM检测核蛋白NF-κB活性。结果支气管滴注LPS成功诱导ALI模型,三叶青黄酮可明显减少BALF中白细胞和中性粒细胞数(P<0.01),降低IL-1β、IL-6、IL-12p40、TNF-α和s TNF-R1的分泌水平(P<0.01),改善肺组织病理损伤。三叶青黄酮明显抑制肺组织p38MAPK、NF-κB的磷酸化及NF-κB的DNA结合活性(P<0.01)。结论三叶青黄酮抑制p38MAPK、NF-κB炎症通路,保护LPS诱导的老年小鼠ALI。     

竹节香附素A抗炎作用的研究北大核心CSCD

目的在构建核转录因子-κB(NF-κB)通路报告基因模型后,研究竹节香附素A(RDA)抗炎的作用及其作用机制。方法用双荧光素酶报告基因技术,以p NF-kappa B-Luc(p NF-κB-Luc)、pr L-tk质粒转染RAW 264.7细胞,脂多糖(LPS)为激活剂,构建靶向NF-κB通路的报告基因模型,RDA和吡咯烷二硫氨基甲酸酯(PDTC)分别给药验证模型。空白组给予空白培养基,模型组给予含LPS的培养基,对照组给予含PDTC的培养基,实验组给予含RDA的培养基。用Western-blot技术检测RDA对细胞蛋白p65和IκBα表达的影响。结果当LPS浓度为10μg·m L^(-1),转染试剂量为2μL时,成功构建靶向NF-κB通路的报告基因模型。RDA能够降低转染后细胞内p65和IκBα亚基的磷酸化,显著抑制NF-κB信号通路。结论本文构建的模型具有快速、稳定、特异、高通量等特点,可用于抗炎活性成分筛选,确认RDA的抗炎机制与NF-κB通路有关。     

The protective effect of terpinen-4-ol on inflammatory injury of VSMCs induced by high glucose through improving autophagic flux disorder北大核心CSCD

Aim To investigate the effect of terpinen-4-ol (T40) on inflammatory injury of vascular smooth muscle cells (VSMCs) induced by high glucose based on the improvement of autophagic flow disorder and involved molecular signals. Methods The scratch test was used to analyze the migration ability of VSMCs, the levels of IL-1β and IL-6 in cell culture supernatant were measured by ELISA, the expression levels of inflammation-related proteins NF-κb p65, p-NF-κb p65, IL-1β, IL-18 and autophagy-related proteins p62, LC3-HYLC3-I, Beclinl, p-Beclinl were de-tected by Western blot. Results T40 inhibited migration of VSMCs induced by high glucose, reduced the secretion and release of pro-inflammatory factors IL-1β and IL-6, inhibited the expression of p-NF-κb p65/ NF-κb p65, IL-1β, IL-18, downregulated the expression of p62, LC3-TJ/LC3- I and p-Beclinl at same time. After interfering the autophagic flux of VSMCs with autophagy inhibitor chloroquine (CQ) , T40 pre-treatment significantly inhibited the protein expression levels of the above inflammatory factors and autophagy-related signals which mediated by CQ. Conclusion T40 inhibits the inflammatory injury of VSMCs induced by high glucose through improving the autophagic flow disorder. © 2023 Publication Centre of Anhui Medical University. All rights reserved.     

Mechanism of Polygonum capitatum in treatment of Helicobacter Pylori associated gastritis by network pharmacology,microarray data analysis and molecular docking北大核心CSCD

Aim To explore the mechanism of Polygonum capitatum(PC)in the treatment of Helicobacter Pylori associated gastritis(HAG). Methods The databases were used to identify the target of PC active compounds and HAG-related genes,and the intersection was taken to obtain the potential targets of PC treatment of HAG. The interaction network diagram of “drug-active compound-target-disease” and the protein-protein interaction(PPI)network of potential target protein interaction in HAG treated by PC were constructed by software Cytoscape 3.6.0. The important nodes in the network were screened by several topological indexes,and the GO and KEGG enrichment were analyzed by STRING database to obtain the potential signaling pathway of PC in the treatment of HAG. The binding ability of PC active components with key target proteins was observed by molecular docking method. On this basis,the related targets of PC in the treatment of HAG were verified in vivo and in vitro experiments. Results The PC active compounds and targets were identified through the database,and the “drug-active compound-target-disease” network diagram and the PPI network of potential target proteins were constructed. Combined with several topological indexes,the PPI network of potential target-protein interaction was analyzed,and 52 hub genes were screened. Further bioinformatics analysis and high-throughput sequencing revealed that PC exerted an effect on HAG through the Akt/NF-κB/NLRP3 pathway. Based on this,it was found that PC could reduce IL-18 and IL-1β in HAG GES-1 cells and HAG SD rats,up-regulate Akt and its phosphorylation level and reduce NF-κB expression,inhibit the activation of NLRP3 inflammatory body,so as to improve HAG inflammatory response. Conclusions PC could exert a therapeutic effect on HAG by activating Akt and its phosphorylation level,and inhibiting the expression of NF-κB and NLRP3 inflammasome related factors. This study provides a theoretical basis for explaining the mechanism of PC in the treatment of HAG. © 2023 Publication Centre of Anhui Medical University. All rights reserved.     

柚皮素对肺炎链球菌诱导的肺泡上皮细胞凋亡及MAPK/NF-κB信号通路的影响

本研究探究柚皮素对肺炎链球菌(SP)诱导的肺泡上皮细胞凋亡及MAPK/NF-κB信号通路的影响。培养肺泡上皮细胞HPAEpiC,使用CCK-8检测柚皮素对HPAEpiC细胞的毒性。再次培养HPAEpiC细胞,分为对照组、模型组、阳性对照组、柚皮素组和抑制剂组,ELISA法检测炎症因子的表达水平;TUNEL检测细胞凋亡水平;Western blot检测凋亡及MAPK/NF-κB信号通路相关蛋白表达水平。结果显示,柚皮素浓度≤60μmol/L时,对HPAEpiC细胞无毒性(P>0.05)。SP诱导可上调细胞上清液中炎症因子(TNF-α和IL-1β)的释放量、细胞凋亡率和细胞中凋亡相关蛋白(Cleaved caspase-8、Cleaved caspase-9和Cleaved caspase-3)与MAPK/NF-κB信号通路相关蛋白(p-JNK、p-p38、p-ERK、p-p65和p-IκBα)的表达水平。柚皮素干预后可改善SP对肺泡上皮细胞的影响,且添加MAPK/NF-κB信号通路抑制剂干预与柚皮素干预对SP诱导的肺泡上皮细胞的作用效果具有相似性。以上结果提示柚皮素可能通过抑制MA...     

枇杷叶提取物通过激活磷脂酰肌醇 3激酶 /蛋白激酶 B信号通路减轻脂多糖诱导的肺细胞损伤

目的探讨枇杷叶提取物对脂多糖( LPS)诱导的人 Ⅱ型肺泡上皮细胞 A549增殖、凋亡及氧化应激的影响及其可能作用机制。方法 2019年 6月至 2020年 7月,采用 LPS诱导的 A549细胞建立肺损伤模型( LPS组)同时将正常培养的细胞作为对照组。不同剂量( 1 mg/L、2 mg/L、4 mg/L)的枇杷叶提取物处理细胞( LPS+枇杷叶 -L组、 LPS+枇,杷叶 -M组、 LPS+枇杷叶 -H组),添加磷脂酰肌醇 3激酶( PI3K)/蛋白激酶 B(Akt)信号通路抑制剂 LY294002与枇杷叶提取物处理细胞( LPS+枇杷叶 -H+ LY294002组);采用 MTT法检测细胞增殖;采用流式细胞术检测细胞凋亡率;采用 2,4-二硝基苯肼显色法检测乳酸脱氢酶(LDH)的含量;采用黄嘌呤氧化酶法检测超氧化物歧化酶(SOD)的含量;采用硫代巴比妥酸法检测丙二醛的含量;蛋白质印迹法( Western blotting)检测磷酸化磷脂酰肌醇 -3-激酶( p-PI3K)、磷酸化蛋白激酶 B(p-Akt)、 B细胞淋巴瘤 -2(Bcl-2)、 Bcl-2相关 X(Bax)蛋白蛋白表达量。结果与对照组比较, LPS组吸光度［( 1.38±0.06)比( 0.55±0.02)］及 SOD的含量［( 81.66±5.36)U/L比(14.65±1.06)U/L］降低,凋亡率［(6.41±0.25)%比( 27.43±1.00)%］、 Bax蛋白水平及 LDH［(242.86±6.09)U/L比( 875.92±15.01)U/ L］、丙二醛［( 121.55±3.17)μmol/g比( 424.46±6.48)μmol/g］的含量升高, Bcl-2、p-PI3K［( 0.60±0.04)比( 0.13±0.01)］、 p-Akt［( 0.51±0.04)比( 0.09±0.01)］蛋白水平降低,差异有统计学意义( P<0.05);与 LPS组比较, LPS+枇杷叶 -M组、 LPS+枇杷叶 -H组吸光度［( 0.55±0.02)比( 0.86±0.03)、(1.18±0.05)］及 SOD［( 14.65±1.06)U/L比( 36.84±2.08)U/L、(70.97±3.03)U/L］的含量升高,凋亡率［( 27.43±1.00)%比( 22.24±0.81)%、(14.78±0.49)%］、 Bax蛋白水平及 LDH［( 875.92±15.01)U/L比( 641.95±9.81)U/L、(365.47±7.02)U/L］、丙二醛［( 424.46±6.48)μmol/g比( 277.94±6.90)μmol/g、(156.30±3.26)μmol/g］的含量降低, Bcl-2、p-PI3K［( 0.13±0.01)比( 0.32±0.01)、(0.54±0.03)］、 p-Akt［( 0.09±0.01)比( 0.25±0.02)、(0.42±0.03)］蛋白水平升高,均差异有统计学意义( P<0.05);添加 LY294002可明显逆转枇杷叶提取物对 LPS诱导的 A549细胞增殖、凋亡及氧化应激的作用。结论枇杷叶提取物可通过激活 PI3K/Akt信号通路减轻 LPS诱导的 A549细胞增殖抑制、凋亡和氧化应激效应,为探究枇杷叶治疗细菌感染     

Synthesis of matrine derivative ZS10 and its mechanism of apoptosis induced by PI3K/AKT signaling pathway in human hepatocellular carcinoma cell line BEL-7402北大核心CSCD

Aim To explore the signaling pathway of matrine derivative ZS10 in inhibiting proliferation and inducing apoptosis of BEL-7402 cells. Methods ZS10 was synthesized by organic synthesis. The inhibitory effect of ZS10 on the proliferation of BEL-7402 cells was analyzed by MTT method at the time of 24 h, 48 h and 72 h, respectively, and IC50 was calculated. DAPI staining was used to observe the state of BEL-7402 cells. Clone formation method was used to observe the colony formation of BEL-7402 cells, flow cytometry was used to observe the cell cycle arrest and apoptosis of BEL7402 cells, and Western blot was used to detect the expression level of PI3K/AKT pathway and related proteins. Results MTT results showed that the IC50 was(6.62±1.11)μmol·L-1; DAPI staining showed that the cell state changed significantly with the increase of drug concentration, and the results of colony formation showed that ZS10 significantly inhibited the colony formation of BEL-7402 cells. The results of flow cytometry showed that ZS10 induced S phase arrest and cycle apoptosis of BEL-7402 cells. Western blot showed that ZS10 at the concentration of 0～8 μ mol·L-1 could regulate the PI3K/AKT pathway and its related proteins in a dose-dependent manner. Compared with the control group, the expression of PI3K, AKT, P-AKT and anti-apoptotic protein Bcl-2 significantly decreased, the expression of pro-apoptotic protein Bax significantly increased, the expression of Cyclin D1 and CDK2 significantly decreased, and the expression of EGFR and N-cadherin, Vimentin significantly decreased in the treatment group. The expression of E-cadherin increased. Conclusions Matrine derivative ZS10 can inhibit the growth and proliferation of hepatocellular carcinoma cell line BEL-7402. © 2023 Publication Centre of Anhui Medical University. All rights reserved.