黄芩多糖对PRV感染猪睾丸细胞的作用及其差异基因表达分析

Effects of Scutellaria baicalensis Georgi Polysaccharide on PRV-infected ST Cells and Analysis of Differential Gene Expression

为研究黄芩多糖对伪狂犬病毒(PRV)感染猪睾丸(ST)细胞的作用并筛选与调控相关的差异基因,本试验建立PRV感染ST细胞模型,将其分为预防、治疗和杀毒模式,分别使用不同浓度的黄芩多糖溶液进行处理,每个模式下设立空白对照组和PRV对照组。采用CCK-8法检测黄芩多糖对ST细胞存活率的影响,显微镜观察各组细胞形态,ELISA检测IFN-γ、IFN-α和TNF-α细胞因子含量;在24 h时提取RNA进行转录组测序,分析其差异表达基因并进行GO功能和KEGG通路分析。结果显示,PRV感染ST细胞出现病变。与PRV对照组相比,黄芩多糖浓度为97.66μg/mL时,3种模式中细胞存活率均极显著升高(P<0.01),细胞病变减轻,IFN-γ和IFN-α含量均极显著提高(P<0.01),TNF-α含量极显著降低(P<0.01),治疗指数(TI)分别为2、4和4。治疗模式下,97.66μg/mL黄芩多糖处理组、空白对照组和PRV对照组比较共交集差异基因74个,其中SOCS3、VEGFA、ZBTB18、CLCN6、RSBN1、RBM47和TET2基因差异表达较为显著,差异表达基因显著富集到15条信号通路,PRV对照组与空白对照组和PRV对照组与黄芩多糖治疗组比较,差异表达基因主要分别涉及核糖体、内吞、MAPK、TNF和MAPK、mTOR、TLR等相关通路。结果表明,黄芩多糖对PRV感染ST细胞具有保护作用,其作用可能是通过调节TNF信号通路和TLR信号通路中SOCS3、CCL5、FOS、PIK3R1和MAP3K8等相关基因的表达来实现。本试验结果将为进一步探索黄芩多糖调控PRV感染宿主的作用机制提供理论依据。

To investigate the effects of Scutellaria baicalensis Georgi polysaccharide(SBGP)on pseudorabies virus(PRV)infection in swine testis(ST)cells and to screen for differentially regulated genes,this study established a PRV-infected ST cell model and divided it into prevention,treatment,and antivirus modes.Different concentrations of SBGP solutions were used for treatment.Each model included blank control groups and PRV control groups.The effects of SBGP on ST cell viability were assessed using the CCK-8 method,and the cell morphology in each group was observed under microscope,the content of cytokines(IFN-γ,IFN-α,and TNF-α)was measured using ELISA.At 24 hours,RNA was extracted for transcriptome sequencing,and differential gene expression was analyzed along with GO function and KEGG pathway analysis.The results showed that PRV infection led to pathological changes in ST cells.Compared with the PRV control group,cell viability significantly increased(P<0.01),cell damage decreased,IFN-γand IFN-αlevels significantly increased(P<0.01),and TNF-αlevels significantly decreased(P<0.01)when treated with 97.66μg/mL of SBGP in all three modes.The treatment index(TI)was 2,4,and 4,respectively.Under the treatment mode,97.66μg/mL SBGP treatment group,blank control group,and PRV control group had 74 overlapping differentially expressed genes.Among these,SOCS 3,VEGFA,ZBTB 18,CLCN 6,RSBN 1,RBM 47,and TET 2 showed significant differential expression.Differentially expressed genes were significantly enriched in 15 signaling pathways.When comparing the PRV control group with the blank control group and the PRV control group with the SBGP treatment group,the differentially expressed genes were mainly associated with ribosomes,endocytosis,MAPK,TNF,MAPK,mTOR,TLR,and other related pathways.These results suggest that SBGP has a protective effect on PRV-infected ST cells,potentially mediated by the regulation of genes such as SOCS 3,CCL 5,FOS,PIK 3 R 1,and MAP 3 K 8 in the TNF and TLR signaling pathways.This study provides a theoretical basis for further exploration of the mechanisms by which SBGP regulates PRV infection in hosts.