目的 測(cè)定去唾液酸糖蛋白受體(ASGPR)在N-二硝基亞乙胺(DEN)誘導(dǎo)大鼠原位肝癌模型(DEN-HCC-Rat)和原位移植大鼠肝癌模型(OTT-HCC-Rat)中的表達(dá)。方法 DEN-HCC-Rat造模:模型組SD大鼠按照20 mg·kg^-1 ig 0.25% DEN水溶液,每周1次,0.025% DEN水溶液供動(dòng)物飲用;對(duì)照組每周ig 1次0.9%氯化鈉溶液,滅菌水供動(dòng)物飲用,于造模第4、10、18、22周取材。OTT-HCC-Rat造模:模型組SD大鼠肝葉注射N1-S1細(xì)胞,假手術(shù)組大鼠麻醉后給予微創(chuàng)縫合處理,于注射后14 d取材。HE染色觀察肝組織病變;通過免疫組化、免疫熒光、Western blotting和實(shí)時(shí)熒光定量PCR(qRT-PCR)實(shí)驗(yàn)檢測(cè)各組ASGPR表達(dá)及分布。結(jié)果 通過HE染色確定DEN-HCC-Rat和OTT-HCC-Rat造模成功。在DEN-HCC-Rat中,免疫組化、免疫熒光、Western blotting、qRT-PCR結(jié)果均顯示,與對(duì)照組比較,模型組大鼠肝組織ASGPR表達(dá)顯著上調(diào)(P＜0.05、0.01),且呈時(shí)間相關(guān)性;在OTT-HCC-Rat中,免疫組化結(jié)果顯示模型組ASGPR表達(dá)顯著高于假手術(shù)組(P＜0.05),免疫熒光、Western blotting、qRT-PCR結(jié)果顯示ASGPR在模型組與假手術(shù)組之間無顯著性差異。結(jié)論 DEN-HCC-Rat模型更好地模擬腫瘤微環(huán)境改變,且ASGPR在肝癌區(qū)域表達(dá)高于對(duì)照組大鼠肝臟,故可利用ASGPR介導(dǎo)肝靶向制劑的轉(zhuǎn)運(yùn),提高肝靶向藥物治療效果。

Objective To determine the role of asialoglycoprotein receptor(ASGPR) in N-dinitroethylamine(DEN)-induced hepatocellular carcinoma model in rats(DEN-HCC-Rat) and orthotopic transplantation tumor mode in rats(OTT-HCC-Rat).Methods DEN-HCC-Rat modeling: SD rats in the model group were ig given a 0.25% DEN aqueous solution of 20 mg·kg^-1, once a week, with 0.025% DEN aqueous solution for animal consumption; The control group received 0.9% sodium chloride solution once a week, sterilized water for animal consumption, and samples were taken at weeks 4, 10, 18, and 22 of modeling. OTT-HCC-Rat modeling: SD rats in the model group were injected with N1-S1 cells into the liver lobes, while rats in the sham surgery group were anesthetized and treated with minimally invasive suturing. Samples were taken 14 days after injection. HE staining was used to observe liver tissue lesions. Immunohistochemistry, immunofluorescence, Western blotting and real-time fluorescent quantitative PCR(qRT-PCR) were used to detect the expression and distribution in the liver tissue of two orthotopic liver cancer rat models and normal rats.Results In DEN-HCC-Rat, immunohistochemistry, immunofluorescence, Western blotting, and qRT-PCR results all showed that compared with the control group, the ASGPR expression in the liver tissue of the model group rats was significantly upregulated(P＜0.05, 0.01), and showed a time correlation. In OTT-HCC-Rat, immunohistochemistry results showed that the expression of ASGPR in the model group was significantly higher than that in the sham surgery group(P＜0.05).Immunofluorescence, Western blotting, and qRT-PCR results showed no significant difference in ASGPR between the model group and the sham surgery group.Conclusion The DEN-HCC-Rat model can better simulate the changes of tumor microenvironment and the expression of ASGPR in liver cancer is higher than that in normal rat liver. Therefore, ASGPR can be used to mediate the transport of liver targeted agents and improve the therapeutic effect of liver targeted drugs.

R965.2

2022年中科院“西部之光”人才培養(yǎng)計(jì)劃