摘要
目的探讨葛黄颗粒含药血清对乙醇诱导的L-02肝细胞损伤的影响及其可能机制。方法SD雄性大鼠45只,随机分为三组(15只/组):空白对照组、葛黄颗粒组及美他多辛组。空白对照组予以蒸馏水灌胃、葛黄颗粒组(3.1 g·100 g^(-1)·d^(-1))、美他多辛组(0.1 g·100 g^(-1)·d^(-1))灌胃,连续干预5天,腹主动脉取血制备含药血清。将L-02肝细胞分为六组并按以下方式干预:正常组(加入10%正常大鼠血清)、模型组(在正常组基础上加入3%乙醇)、葛黄颗粒高、中、低剂量组(分别加入不同浓度葛黄颗粒含药血清及正常大鼠血清及3%乙醇)、美他多辛组(加入3%乙醇、美他多辛及正常大鼠血清各5%),将L-02肝细胞培养24 h后进行之后实验部分。ELISA法检测NF-κB、TNF-α、IL-1β、IL-18水平;RT-PCR法检测NLRP3、ASC、Caspase-1 mRNA的表达;Western blot法检测TLR4、NLRP3、ASC、Caspase-1的蛋白表达。结果与正常组比较:模型组NF-κB、TNF-α、IL-1β、IL-18明显升高,NLRP3、ASC、Caspase-1 mRNA表达明显升高,TLR4、NLRP3、ASC、Caspase-1的蛋白表达均亦明显升高(P<0.05);与模型组相比较:以上指标在葛黄颗粒高、中剂量组及美他多辛组表达水平均有所降低(P<0.05);与美他多辛组比较:葛黄颗粒高剂量组中TNF-α、IL-18有统计学意义(P<0.05),而IL-1β、NF-KB差异则不显著(P>0.05);葛黄颗粒低剂量组中NLRP3、ASC、Caspase-1 mRNA差异显著(P<0.05),而在高、中剂量组差异则均无统计学意义(P>0.05);TLR4、NLRP3、ASC、Caspase-1的蛋白表达水平在葛黄颗粒各剂量组均有统计学意义(P<0.05)。结论葛黄颗粒含药血清可能通过TLR4/NLRP3信号通路改善乙醇诱导的肝细胞炎症损伤。
Objective To investigate the effect of serum containing Gehuang granules on ethanol-induced L-02hepatocytes injury and its possible mechanism.Methods 45 SD male rats were randomly divided into three groups(15 rats/group):blank control group,Gehuang granule group and Metadoxine group.The blank control group was intragastrically administered with distilled water,Gehuang Granule group(3.1 g·100 g^(-1)·d^(-1))and Metadoxine group(0.1 g·100 g^(-1)·d^(-1))for 5 days.The blood was collected from abdominal aorta to prepare drug-containing serum.The L-02 hepatocytes were divided into six groups and intervened according to the following methods:normal group(adding 10%normal rat serum),model group(adding 3%ethanol on the basis of normal group),Gehuang granules high,medium and low dose groups(adding different concentrations of Gehuang granules medicated serum and normal rat serum and 3%ethanol respectively),and Metadoxine group(adding 3%ethanol,Metadoxine and normal rat serum 5%each).The L-02 hepatocytes were cultured for 24 h and then the experimental part was carried out.The levels of NF-κB,TNF-α,IL-1βand IL-18 were detected by ELISA.RT-PCR was used to detect the expression of NLRP3,ASC and Caspase-1.The protein expressions of TLR4,NLRP3,ASC and Caspase-1 were detected by Western blot.Results Compared with the normal group,the expressions of NF-κB,TNF-α,IL-1β,IL-18,and the expression of NLRP3,ASC,Caspase-1mRNA and the levels of TLR4,NLRP3,ASC and caspase-1 protein in the model group were significantly increased(P<0.05).Compared with the model group,the expression levels of the above indexes in Gehuang granules high,medium dose group and Metadoxine group were decreased(P<0.05).Compared with the Metadoxine group:Gehuang granules high dose group TNF-α,IL-18 was statistically significant(P<0.05),However,there was no significant difference between high and medium dose groups(P>0.05).NLRP3,ASC,Caspase-1mRNA in Gehuang granules low dose group difference was statistically significant(P<0.05),however,IL-1β,NF-κB differ
作者
吴莉
李波
王晓栋
康莉
刘友平
魏嵋
Wu Li;Li Bo;Wang Xiaodong;Kang Li;Liu Youping;Wei Mei(The Affiliated TCM Hospital of Southwest Medical University,Luzhou 646000,China;School of Basic Medicine,Southwest Medical University,Luzhou 646000,China)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2022年第10期3828-3834,共7页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
四川省教育厅重点项目(16ZA0193):葛黄颗粒对大鼠酒精性肝损伤的防治作用及其分子机制研究,负责人:魏嵋。
