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萝卜硫素对坏死性小肠结肠炎小鼠内质网应激诱导的细胞凋亡和炎症的保护作用

The Protective Effect of Sulforaphane on ER-induced Apoptosis and Inflammation in Necrotizing Enterocolitis Mice.

作者信息

Wang Xinhong, Mi Yanhong, Xiong Xiaoyu, Bao Zhongkun

机构信息

Department of Radiology, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China.

Department of Radiology, Children's Hospital of Zhejiang University, Hangzhou, Zhejiang Province, China.

出版信息

Comb Chem High Throughput Screen. 2023;26(6):1186-1195. doi: 10.2174/1386207325666220705115007.

Abstract

BACKGROUND

Necrotizing enterocolitis (NEC) is a neonatal intestinal necrotizing disease caused by various factors in newborns. Sulforaphane (SFN) has a strong anti-inflammatory ability and a certain protective effect on intestinal diseases.

OBJECTIVE

NEC is a common developed gastrointestinal exigency in an untimely baby. SFN is a naturally originated isothiocyanate that has beneficial effects on the intestinal system.The purpose of this study is to study the protective effect of SFN on endoplasmic reticulum stress (ERS)-related NEC.

METHODS

The newborn mice were randomly divided into control (n = 15), NEC (n = 20), and NEC+SFN (n = 18) groups. Mice in NEC and SFN+NEC groups were injected with 0.1 μl normal saline or 20 mg/kg/d SFN, respectively. After that, the weight and survival of the mice were recorded every day. Then the mice were sacrificed after 96 h of modeling; ileum tissue and blood samples were collected for qPCR, Western blot, ELISA, HE staining, TUNEL staining, and immunohistochemistry assays.

RESULTS

SFN significantly inhibited the mRNA expression of BIP, CHOP, IL-1β and IL-6, and protein expression of Bax, Caspase-3, Caspase-9 and CHOP, and promoted the expression of Bcl-2 in ER-induced NEC mice intestinal tissues (P<0.01). Meanwhile, SFN could suppress the serum levels of IL-8, IL-10, IL-6, TNF-α, and IL-1β, and positive expression of TLR4 and NF-κB (P<0.01), and promote the serum levels of IL-10. HE staining showed that SFN alleviated the NEC intestinal tissue injury, and TUNNEL staining showed that SFN could reduce the rate of NEC apoptotic cells (P<0.01). Moreover, SFN treatment improved the body weight and survival rate in NEC mice.

CONCLUSION

SFN could effectively protect against ERS-induced inflammation and apoptosis in NEC mice.

摘要

背景

坏死性小肠结肠炎(NEC)是一种由多种因素引起的新生儿肠道坏死性疾病。萝卜硫素(SFN)具有较强的抗炎能力,对肠道疾病有一定的保护作用。

目的

NEC是早产儿常见的严重胃肠道急症。SFN是一种天然存在的异硫氰酸盐,对肠道系统有益。本研究旨在探讨SFN对内质网应激(ERS)相关NEC的保护作用。

方法

将新生小鼠随机分为对照组(n = 15)、NEC组(n = 20)和NEC+SFN组(n = 18)。NEC组和SFN+NEC组小鼠分别注射0.1 μl生理盐水或20 mg/kg/d SFN。此后,每天记录小鼠的体重和存活情况。建模96小时后处死小鼠;收集回肠组织和血液样本进行qPCR、蛋白质免疫印迹、酶联免疫吸附测定、苏木精-伊红染色、脱氧核糖核苷酸末端转移酶介导的缺口末端标记染色和免疫组织化学分析。

结果

SFN显著抑制ER诱导的NEC小鼠肠道组织中BIP、CHOP、IL-1β和IL-6的mRNA表达,以及Bax、Caspase-3、Caspase-9和CHOP的蛋白表达,并促进Bcl-2的表达(P<0.01)。同时,SFN可抑制血清中IL-8、IL-10、IL-6、TNF-α和IL-1β水平,以及TLR4和NF-κB的阳性表达(P<0.01),并促进血清IL-10水平。苏木精-伊红染色显示SFN减轻了NEC肠道组织损伤,脱氧核糖核苷酸末端转移酶介导的缺口末端标记染色显示SFN可降低NEC凋亡细胞率(P<0.01)。此外,SFN治疗提高了NEC小鼠的体重和存活率。

结论

SFN可有效预防ERS诱导的NEC小鼠炎症和凋亡。

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