脱氧紫草素通过SIRT1/FoxO1/Rab7轴调节自噬和氧化应激来抑制轮状病毒复制。

Deoxyshikonin inhibited rotavirus replication by regulating autophagy and oxidative stress through SIRT1/FoxO1/Rab7 axis.

作者信息

Huang Haohai, Liao Dan, He Bin, Pu Rong, Cui Yejia, Zhou Guanghui

机构信息

Medical and Pharmacy Research Laboratory, SSL Central Hospital of Dongguan, Dongguan Third People's Hospital, Affiliated Dongguan Shilong People's Hospital of Southern Medical University, Dongguan, Guangdong, China; Department of Clinical Pharmacy, SSL Central Hospital of Dongguan, Dongguan Third People's Hospital, Affiliated Dongguan Shilong People's Hospital of Southern Medical University, Dongguan, Guangdong, China.

Medical and Pharmacy Research Laboratory, SSL Central Hospital of Dongguan, Dongguan Third People's Hospital, Affiliated Dongguan Shilong People's Hospital of Southern Medical University, Dongguan, Guangdong, China; Department of Gynaecology, SSL Central Hospital of Dongguan, Dongguan Third People's Hospital, Affiliated Dongguan Shilong People's Hospital of Southern Medical University, Dongguan, Guangdong, China.

出版信息

Microb Pathog. 2023 May;178:106065. doi: 10.1016/j.micpath.2023.106065. Epub 2023 Mar 11.

DOI:10.1016/j.micpath.2023.106065

PMID:36907361

Abstract

BACKGROUND

Rotavirus (RV) is a double-stranded RNA virus. RV prevention and treatment remain a major public health problem due to the lack of clinically specific drugs. Deoxyshikonin is a natural compound isolated from the root of Lithospermum erythrorhizon and one of the shikonin derivatives which owns remarkable therapeutic effects on multiple diseases. The purpose of this research was to inquire Deoxyshikonin's role and mechanism in RV infection.

METHODS

Deoxyshikonin's function in RV was estimated using Cell Counting Kit-8 analysis, cytopathic effect inhibition assay, virus titer determination, quantitative real-time PCR, enzyme linked-immunosorbent assay, Western blot, immunofluorescence, and glutathione levels detection. Also, Deoxyshikonin's mechanism in RV was appraised with Western blot, virus titer determination, and glutathione levels detection. Moreover, Deoxyshikonin's function in RV in vivo was determined using animal models, and diarrhea score analysis.

RESULTS

Deoxyshikonin owned anti-RV activity and repressed RV replication in Caco-2 cells. Furthermore, Deoxyshikonin reduced autophagy and oxidative stress caused by RV. Mechanistically, Deoxyshikonin induced low protein levels of SIRT1, ac-Foxo1, Rab7, VP6, low levels of RV titers, low autophagy and oxidative stress. SIRT1 overexpression abolished the effects of Deoxyshikonin on RV-treated Caco-2 cells. Meanwhile, in vivo research affirmed that Deoxyshikonin also possessed anti-RV function, and this was reflected in increased survival rate, body weight, GSH levels, and decreased diarrhea score, RV virus antigen, LC-3II/LC3-I.

CONCLUSION

Deoxyshikonin reduced RV replication through mediating autophagy and oxidative stress via SIRT1/FoxO1/Rab7 pathway.

摘要

背景

轮状病毒（RV）是一种双链RNA病毒。由于缺乏临床特效药物，RV的预防和治疗仍然是一个重大的公共卫生问题。脱氧紫草素是从紫草根部分离出的一种天然化合物，是紫草素衍生物之一，对多种疾病具有显著的治疗作用。本研究的目的是探究脱氧紫草素在RV感染中的作用及机制。

方法

使用细胞计数试剂盒-8分析、细胞病变效应抑制试验、病毒滴度测定、定量实时PCR、酶联免疫吸附测定、蛋白质免疫印迹法、免疫荧光法和谷胱甘肽水平检测来评估脱氧紫草素在RV中的作用。此外，通过蛋白质免疫印迹法、病毒滴度测定和谷胱甘肽水平检测来评估脱氧紫草素在RV中的作用机制。此外，使用动物模型和腹泻评分分析来确定脱氧紫草素在体内对RV的作用。

结果

脱氧紫草素具有抗RV活性，并能抑制Caco-2细胞中RV的复制。此外，脱氧紫草素可减轻RV引起的自噬和氧化应激。机制上，脱氧紫草素可诱导SIRT1、乙酰化Foxo1、Rab7、VP6的蛋白水平降低，RV滴度降低，自噬和氧化应激水平降低。SIRT1过表达消除了脱氧紫草素对RV处理的Caco-2细胞的影响。同时，体内研究证实脱氧紫草素也具有抗RV功能，这体现在存活率、体重、谷胱甘肽水平增加，腹泻评分、RV病毒抗原、LC-3II/LC3-I降低。