摘要
目的探讨黄芪多糖(APS)对急性放射性肠炎(ARE)大鼠肠黏膜细胞凋亡及TXNIP/NLRP3轴的影响。方法按照随机数字表法将大鼠分为6组,即对照组(NC组)、模型组(Model组)、黄芪多糖低剂量组(APS-L组)、黄芪多糖中剂量组(APS-M组)、黄芪多糖高剂量组(APS-H组)、阳性药物组(Positive组),每组12只。除NC组外,其余各组大鼠均构建ARE模型,造模24 h后,APS-L组、APS-M组、APS-H组大鼠分别按100、200、400 mg·kg^(-1)APS的剂量进行灌胃,Positive组大鼠按照300 mg·kg^(-1)柳氮磺吡啶的剂量进行灌胃,NC组及Model组大鼠给予生理盐水灌胃,末次给药24 h后,对各组大鼠进行疾病活动指数(DAI)评分;苏木精-伊红(HE)染色观察各组大鼠肠黏膜病理学变化;ELISA法检测各组大鼠肠黏膜中IL-6、TNF-α的含量;TUNEL法检测肠黏膜细胞凋亡;qRT-PCR检测大鼠肠黏膜组织中TXNIP、NLRP3、Caspase-1、IL-1β、IL-18 mRNA的表达;Western blot检测各组大鼠肠黏膜组织中Bax、半胱氨酸天冬氨酸蛋白水解酶3(Caspase-3)及TXNIP/NLRP3通路相关蛋白的表达。结果与NC组比较,Model组大鼠肠黏膜损伤严重,出现炎性细胞浸润,隐窝及上皮破坏等现象,DAI评分、白细胞介素6(IL-6)含量、肿瘤坏死因子-α(TNF-α)含量、细胞凋亡率及Bax蛋白、Caspase-3蛋白、TXNIP、NLRP3、Caspase-1、IL-1β、IL-18 mRNA及蛋白表达水平显著升高(P<0.05);与Model组比较,APS-L组、APS-M组、APS-H组及Positive组大鼠肠黏膜损伤程度明显减轻,DAI评分、IL-6含量、TNF-α含量、细胞凋亡率及Bax蛋白、Caspase-3蛋白、TXNIP、NLRP3、Caspase-1、IL-1β、IL-18 mRNA及蛋白表达水平显著降低(P<0.05)。结论APS可抑制ARE大鼠肠黏膜细胞凋亡,该机制可能与抑制TXNIP/NLRP3轴有关。
Objective To determine the effect of astragalus polysaccharide(APS)on the apoptosis and TXNIP/NLRP3 axis of the intestinal mucosal cells in rats with acute radiation enteritis(ARE).Methods According to the random number table,the rats were divided into 6 groups:a control group(NC group),a model group(Model group),a low-dose astragalus polysaccharide group(APS-L group),a middledose astragalus polysaccharide group(APS-M group),a high-dose astragalus polysaccharide group(APS-H group),and a positive drug group(Positive group),with 12 rats in each group.Except for the NC group,ARE models were constructed the rats in other groups.After 24 h of modeling,the rats in the APS-L group,the APS-M group,and the APS-H group were given 100,200,and 400 mg·kg^(-1)APS by gavage,respectively.The rats in the Positive group were given 300 mg·kg^(-1)sulfasalazine by gavage,and the rats in the NC group and Model group were given normal saline by gavage.At 24 h after the last administration,the disease activity index(DAI)score was measured on rats in each group.Hematoxylineosin(HE)staining was used to observe the pathological changes of the intestinal mucosa of rats in each group.ELISA was used to detect the levels of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in the intestinal mucosa of rats in each group.TUNEL was used to detect the intestinal mucosal cell apoptosis.qRT-PCR was used to detect the mRNA expressions of TXNIP,NLRP3,Caspase-1,IL-1βand IL-18 in the rat intestinal mucosa.Western blot was used to detect the expression of Bax,Caspase-3 and TXNIP/NLRP3 pathway related proteins in the intestinal mucosal tissues of the rats in each group.Results Compared with the NC group,the intestinal mucosa was severely damaged,with inflammatory cell infiltration,crypt and epithelium destruction.The DAI score,IL-6 content,TNF-αcontent,apoptosis rate and Bax protein,Caspase-3 protein,TXNIP,NLRP3,Caspase-1,IL-1β,IL-18 mRNA and protein expression levels were significantly increased in the Model group(P<0.05).Compared with the Model group,while the intestinal mucosal damage was significantly reduced,the DAI score,IL-6 content,TNF-αcontent,apoptosis rate and Bax protein,Caspase-3 protein,TXNIP,NLRP3,Caspase-1,IL-1β,IL-18 mRNA and protein expression levels were significantly increased in the APS-L group,the APS-M group,the APS-H group and the Positive group(P<0.05).Conclusion APS can inhibit the intestinal mucosal cell apoptosis in ARE rats,and the mechanism may be related to the inhibition of TXNIP/NLRP3 axis.
作者
李浩田
李鹏
王玉凤
张春礼
LI Hao-tian;LI Peng;WANG Yu-feng;ZHANG Chun-li(Department of General Surgery,Zhengzhou Ninth People’s Hospital,Zhengzhou 450053;Second Department of General Surgery,People’s Hospital of Henan University of Traditional Chinese Medicine/Zhengzhou People’s Hospital,Zhengzhou 450053)
出处
《中南药学》
CAS
2023年第5期1256-1262,共7页
Central South Pharmacy
基金
2019年度河南省医学科技攻关计划(联合共建)项目(No.LHGJ20191072)。
关键词
黄芪多糖
急性放射性肠炎
肠黏膜
细胞凋亡
硫氧还蛋白互作蛋白/炎性小体3通路
astragalus polysaccharide
acute radiation enteritis
intestinal mucosa
apoptosis
thioredoxin interaction protein/NLR family pyrin domain containing 3 pathway
作者简介
李浩田,男,副主任医师,主要从事普外(肝胆,胃肠,甲乳,周围血管,创面)疾病研究,email:sh19work@163.com;通信作者:张春礼,男,副主任医师,主要从事胃肠外科研究,email:justzhang@126.com。
