[1]廖炜波,鲁弘骎,梁浩,等.乙酰紫草素对HCV复制的影响及其作用机制研究▲[J].内科,2019,14(02):137-141.[doi:10.16121/j.cnki.cn45-1347/r.2019.02.04]
 LIAO Weibo,LU Hongqin,LIANG Hao,et al.Research on the effects of acetyl shikonin on HCV replication and its mechanism(s)[J].Internal Medicine of China,2019,14(02):137-141.[doi:10.16121/j.cnki.cn45-1347/r.2019.02.04]
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乙酰紫草素对HCV复制的影响及其作用机制研究▲()
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《内科》[ISSN:1006-6977/CN:61-1281/TN]

卷:
14卷
期数:
2019年02
页码:
137-141
栏目:
论著
出版日期:
2019-04-30

文章信息/Info

Title:
Research on the effects of acetyl shikonin on HCV replication and its mechanism(s)
文章编号:
1673-7768(2019)02-0137-05
作者:
廖炜波1鲁弘骎1梁浩2叶力2陈晖1*
1 广西医科大学第一附属医院;2 广西艾滋病防治研究重点实验室,南宁市530021
Author(s):
LIAO Weibo1 LU Hongqin1 LIANG Hao2 YE Li2 CHEN Hui1
1 The First Affiliated Hospital of Guangxi Medical University; 2 Guangxi Key Laboratory of AIDS Prevention and Treatment, Nanning 530021, China
关键词:
丙型肝炎病毒乙酰紫草素细胞自噬Huh7细胞
Keywords:
Hepatitis C virus Acetyl shikonin Autophagy Huh7 cells
分类号:
R 512.6;R 373.2
DOI:
10.16121/j.cnki.cn45-1347/r.2019.02.04
文献标志码:
A
摘要:
目的探讨乙酰紫草素对HCV复制的影响及其作用机制。方法(1)体外培养Huh7细胞,用不同浓度乙酰紫草素溶液与Huh7细胞进行共同培养,采用MTS法检测细胞的相对活力,观察乙酰紫草素对Huh7细胞的毒性作用。(2)采用JFH-1毒株感染Huh7细胞后,用不同浓度(0、0.1、0.2、0.4、0.8 μM)的乙酰紫草素溶液进行处理,检测HCV RNA以及HCV core蛋白的表达水平,观察乙酰紫草素对Huh7细胞中HCV JFH-1复制的影响。(3)取JFH-1感染的Huh7细胞分为HCV组和HCV+乙酰紫草素组,HCV+乙酰紫草素组用0.8 μM乙酰紫草素处理,HCV组不处理;取未感染JFH-1的Huh7细胞,分为乙酰紫草素组和对照组,乙酰紫草素组用0.8 μM乙酰紫草素处理,对照组不处理。通过检测LC3 mRNA以及LC3B蛋白的表达水平,观察乙酰紫草素对细胞自噬标志因子LC3表达的影响。结果(1)乙酰紫草素浓度在1.0 μM或更低时对Huh7细胞生长影响很小或没有影响,在2.50 μM以上时表现出明显的细胞毒性作用。(2)不同浓度乙酰紫草素(0.1、0.2、0.4、0.8 μM)处理组细胞内HCV RNA水平均显著低于未用乙酰紫草素处理组,差异有统计学意义(P<0.05),HCV core蛋白的表达量均显著低于未用乙酰紫草素处理组。(3)在基因水平上,LC3 mRNA的表达水平表现为HCV组>对照组>HCV+乙酰紫草素组>乙酰紫草素组,差异有统计学意义(P<0.05);在蛋白水平上,LC3B蛋白的表达水平表现为HCV组>对照组>HCV+乙酰紫草素组>乙酰紫草素组。结论乙酰紫草素可以抑制HCV复制,同时下调自噬标志因子LC3的表达,可通过调节细胞自噬途径影响HCV病毒的复制。
Abstract:
ObjectiveTo investigate the effects of acetyl shikonin on HCV replication and its mechanism(s). Methods(1) Huh7 cells were cultured in vitro and co-culture with different concentrations of acetyl shikonin solution. MTS cell proliferation assay was used to detect the relative viability of the cells and the cytotoxic effect of acetyl shikonin on Huh7 cells was observed. (2)After infected with JFH-1 strain, Huh7 cells were treated with different concentrations (0, 0.1, 0.2, 0.4, 0.8 μM) of acetyl shikonin solution. The expression levels of HCV RNA and HCV core protein were detected and the effect of acetyl shikonin on HCV JFH-1 replication in Huh7 cells was observed. (3)Huh7 cells infected with JFH-1 were divided into HCV group and HCV+acetyl shikonin group. The HCV+acetyl shikonin group was treated with 0.8 μM acetyl shikonin, while the HCV group was not treated with acetyl shikonin. Huh7 cells noninfected with JFH-1 were divided into acetyl shikonin group and control group. The acetyl shikonin group was treated with 0.8 μM acetyl shikonin, and the control group was not treated with acetyl shikonin. The effect of acetyl shikonin on the expression of autophagy marker LC3 was observed by detecting the expression levels of LC3 mRNA and LC3B protein. Results(1)Acetyl shikonin had little or no effect on the growth of Huh7 cells at the concentration ≤1.0 μM, and showed a significant cytotoxic effect at the concentration≥2.50 μM. (2)The levels of HCV RNA in the groups treated with different concentrations of acetyl shikonin (0.1, 0.2, 0.4, 0.8 μM) were significantly lower than those in the group untreated with acetyl shikonin, the difference was statistically significant (P<0.05), the expression level of HCV core protein was significantly lower than that in the group untreated with acetyl shikonin. (3)At the gene level, the mRNA expression level of LC3 was HCV group >control group>HCV+acetyl shikonin group>acetyl shikonin group, the difference was statistically significant (P<0.05). At the protein level, the protein expression level of LC3B was HCV group>control group>HCV+acetyl shikonin group>acetyl shikonin group. ConclusionAcetyl shikonin can inhibit HCV replication and down-regulate the expression of the autophagy marker LC3, acetyl shikonin can affect the replication of HCV virus by regulating the autophagy pathway.

相似文献/References:

[1]蒙江明.HIV和HCV混合感染的抗病毒治疗[J].内科,2010,(04):421.
 [J].Internal Medicine of China,2010,(02):421.

备注/Memo

备注/Memo:
▲基金项目:国家自然科学基金资助项目(81460305)
 *通信作者:陈晖,广西医科大学第一附属医院,电子邮箱 chenhuiyfy@gxmu.edu.cn
更新日期/Last Update: 2019-05-09