藤黄酸对脓毒症大鼠肠道损伤的保护作用  

Effect of Gambogic Acid on Intestinal Injury in Septic Rats

作　　者：曾静[1] 杨舒雅[1] 钱娴静 张清[1] 陈娇 敬媛媛 汪耘吉 张栋珉 胥勋梅[1] ZENG Jing;YANG Shu-ya;QIAN Xian-jing;ZHANG Qing;CHEN Jiao;JING Yuan-yuan;WANG Yun-ji;ZHANG Dong-min;XU Xun-mei(Department of Endocrinology,the Third People’s Hospital of Mianyang City,Mianyang 621100,China)

机构地区：[1]绵阳市第三人民医院内分泌科,四川绵阳621100

出　　处：《南昌大学学报（医学版）》2024年第2期23-27,34,共6页Journal of Nanchang University：Medical Sciences

基　　金：绵阳市第三人民医院2022年科研立项课题(202207)。

摘　　要：目的探讨藤黄酸(GA)对脓毒症大鼠肠损伤的影响以及对RIP1/RIP3/MLKL信号通路的调节作用。方法取40只SPF级SD大鼠,按随机数字表法分为假手术(Sham)组、脓毒症(Spesis)组、低剂量GA组和高剂量GA组,各10只。除Sham组外,其余3组大鼠均采用盲肠穿孔结扎法(CLP)建立脓毒症大鼠模型。低剂量GA组和高剂量GA组于造模前24 h分别灌胃给予50、100 mg·kg^(-1)GA。造模后12 h麻醉处死,收集外周血和小肠组织。采用酶联免疫吸附(ELISA)法检测血清中肿瘤坏死因子-α(TNF-α)、白介素1β(IL-1β)和白介素6(IL-6)表达水平;苏木素-伊红(HE)染色观察小肠组织病理变化并进行Chui’s评分;蛋白免疫印迹(Western blotting)法检测凋亡相关蛋白(Caspase-3、Bax和Bcl2)、紧密连接蛋白(ZO-1和Occludin)及RIP1/RIP3/MLKL信号通路相关蛋白的表达水平。结果与Sham比较,Spesis组大鼠肠黏膜受损明显,Chui’s评分显著升高(P<0.05);血清中炎性因子TNF-α、IL-1β、和IL-6表达明显增加(P<0.05);小肠组织中Bax、ZO-1和Occludin蛋白表达明显降低(P<0.05),而Caspase-3、Bcl2、MLKL、RIP1以及RIP3蛋白表达明显增加(P<0.05);与Spesis组比较,低剂量GA组和高剂量GA组大鼠肠黏膜损伤均有不同程度改善,Chui’s评分均明显降低(P<0.05);血清中炎性因子TNF-α、IL-1β、和IL-6表达均显著降低(P<0.05);小肠组织中Bax、ZO-1和Occludin蛋白表达明显增加(P<0.05),而Caspase-3、Bcl2、MLKL、RIP1以及RIP3蛋白表达明显降低(P<0.05)。结论GA能够减轻脓毒症诱导的大鼠肠道损伤,这或许与RIP1/RIP3/MLKL信号通路密切相关。Objective To investigate the effect of Gambogic Acid(GA)on intestinal injury in septic rats.Methods A total of 40 SPF SD rats were divided into Sham group,Sepsis group,Low-dose GA group and High-dose GA group according to the randomized numerical table method,10 rats in each group.The rat models of sepsis were established by cecal perforation ligation(CLP)in all three groups of rats except Sham group.Low-dose GA group and high-dose GA group were respectively given 50 and 100 mg·kg^(-1)GA by gavage 24 h before modeling,and were anesthetized and executed 12 h after modeling,and peripheral blood and small intestinal tissues were collected.The expression levels of tumor necrosis factor-α(TNF-α),interleukin 1β(IL-1β)and interleukin-6(IL-6)in serum were detected by enzyme-linked immunosorbent assay(ELISA);hematoxylin-eosin(HE)staining was used to observe the pathological changes in the small intestinal tissues and Chui’s score were performed;Western blotting was used to detect the relative expression levels of apoptosis-related proteins(Caspase-3,Bax and Bcl2),tight junction proteins(ZO-1 and Occludin)and proteins related to RIP1/RIP3/MLKL signaling pathway.Results Compared with Sham group,the intestinal mucosa of the rats in Spesis group was significantly destroyed,and Chui’s score was up-regulated(P<0.05).Also,the levels of inflammatory factors(TNF-α,IL-1βand IL-6)in serum were significantly increased(P<0.05);the relative expression levels of anti-apoptosis protein Bax and tight junction protein zona(Claudin-1 and ZO-1)in small intestine tissue was significantly decreased(P<0.05),while the relative expression levels of pro-apoptosis protein(Caspase-3 and Bcl2),and RIP1/RIP3/MLKL pathway-associated protein(MLKL,RIP1 and RIP3)were significantly increased(P<0.05).Compared with Spesis group,the intestinal mucosal injury of rats in both Low-dose GA group and High-dose GA group showed different degrees of improvement,and Chui’s score was down-regulated(P<0.05).The levels of inflammatory factors(TNF-α,IL-1βan

关 键 词：藤黄酸 脓毒症 肠道损伤 炎症反应 信号通路 动物 实验 大鼠 

分 类 号：R-332[医药卫生]