Transcriptome profiling of ASPP2-konckdown mouse liver were analyzed by RNA-sequencing. HBV-ASPP2-knockdown mice was the hybrid offspring of HBV transgenic mice and ASPP2 knockdown mice. Liver tissues were taken for the experiments such as western Blot (WB), PCR, Hematoxylin and Eosin (HE), Immunohistochemistry and high throughput sequencing of transcriptome.
Pathological and transcriptomic analysis of liver tissue from ASPP2 knockdown vs con mice showed that after ASPP2 knockdown, the pathological changes in the liver tissue of mice were not significant, but transcriptomics showed obvious changes in immune system process, and response to stimulus, metabolism, Human Diseases and other directions etc. In the HBV-ASPP2-knockdown mice, liver tissue HE staining found less cell swelling and necrosis foci; F4/80 and MPO staining showed less inflammatory cell infiltration; serum ALT and AST decreased than the HBV-ASPP2-con mice. Transcriptome results showed significantly changed in HBV-ASPP2-knockdown mice including immune system process, inflammatory response, and innate immune response etc. Further comparison of the two transcriptomes yielded 9 identical pathways related to inflammatory and cell injury. The PPAR pathway was verified, and found that the increase of PPARγ caused by the reduction of ASPP2 is likely to be the reason for the reduction of HBV-related liver injury. The expression of PPARγ was then analyzed by transcriptome and PCR, it was found that in the absence of HBV, ASPP2 knockdown resulted in a mild decrease in PPARγ, and in the presence of HBV infection, ASPP2 knockdown resulted in a marked increase in PPARγ.In addition, this study found that high expression of ASPP2 had opposite effects on HCC (HBV-none) and HCC (HBV-yes).
This study demonstrated that reduction of ASPP2 reduces HBV-induced hepatocyte damage during chronic HBV infection. This phenomenon is related to the different regulation of PPARγ by ASPP2 in the presence or absence of HBV stimulation.
通过RNA测序分析ASPP2-konckdown小鼠肝脏的转录组谱分析。HBV-ASPP2敲低小鼠是HBV转基因小鼠和ASPP2敲低小鼠的杂交后代。取肝脏组织进行实验,如WesternBlot(WB),PCR,苏木精和伊红(HE),转录组的免疫组织化学和高通量测序。
ASPP2敲低与对照小鼠肝组织的病理学和转录组学分析显示,在ASPP2敲低后,小鼠肝脏组织的病理变化不显著,但是转录组学显示了免疫系统过程的明显变化,对刺激的反应,新陈代谢,人类疾病和其他方向等。在HBV-ASPP2敲低小鼠中,肝组织HE染色发现细胞肿胀和坏死灶较少;F4/80和MPO染色显示炎症细胞浸润较少;血清ALT和AST比HBV-ASPP2-con小鼠降低。转录组结果显示HBV-ASPP2-knockdown小鼠包括免疫系统过程中显著改变,炎症反应,和先天免疫反应等。两个转录组的进一步比较产生了与炎症和细胞损伤相关的9个相同的途径。PPAR途径得到验证,并发现ASPP2降低引起的PPARγ升高很可能是HBV相关性肝损伤减轻的原因。然后通过转录组和PCR分析PPARγ的表达,发现在没有HBV的情况下,ASPP2敲低导致PPARγ轻度降低,在HBV感染的情况下,ASPP2敲低导致PPARγ显著增加。此外,这项研究发现,ASPP2的高表达对HCC(HBV-none)和HCC(HBV-yes)有相反的作用。
这项研究表明,ASPP2的减少减少慢性HBV感染期间HBV诱导的肝细胞损伤。这种现象与在存在或不存在HBV刺激的情况下ASPP2对PPARγ的不同调节有关。