基于NLRP3/Caspase-1/GSDMD通路探讨阿霉素纳米凝胶载药对H22肝癌荷瘤小鼠的作用机制  

Mechanism of Doxorubicin Nanogel Loading on H22Liver Cancer-bearing Mice Based on NLRP3/Caspase-1/GSDMD Pathway

作　　者：张书文 崔佳乐[1] 邱春辉 刘腾予 黄可欣[1] ZHANG Shuwen;CUI Jiayue;QIU Chunhui;LIU Tengyu;HUANG Kexin(Basic Medical College,JilinUniversity,Changchun,Jilin 130021,China)

机构地区：[1]吉林大学基础医学院,吉林长春130021

出　　处：《中国实验诊断学》2025年第1期74-78,共5页Chinese Journal of Laboratory Diagnosis

基　　金：吉林大学“大学生创新创业训练计划”(S202310183585)。

摘　　要：目的通过检测NOD样受体蛋白3(NLRP3)/半胱氨酸天冬氨酸蛋白水解酶-1(Caspase-1)/消皮素D(GSDMD)通路,探讨阿霉素纳米凝胶载药(Nanogel drug loading,NG)对H22肝癌荷瘤小鼠的作用机制。方法将H22肝癌细胞珠每只1.25×10^(6)个/100μl接种于Balb/c小鼠右前肢腋窝皮下,待肿瘤体积至50~60mm^(3)时,将40只Balb/c小鼠随机分为5组(n=8),分别为肿瘤对照组、阿霉素3mg组(DOX-3.0)、阿霉素6mg组(DOX-6.0)、阿霉素纳米凝胶载药3mg组(NG/DOX-3.0)和阿霉素纳米凝胶载药6mg组(NG/DOX-6.0)。各组均采用尾静脉注射给药治疗,分别给予生理盐水(5ml/kg)、DOX(3mg/kg)、DOX(6mg/kg)、NG/DOX(3mg/kg)和NG/DOX(6mg/kg),1次/3d,给药4次,于给药结束第3d处死动物取材。通过苏木精-伊红(hematoxylin-eosin,HE)染色,观察肿瘤生长情况和坏死面积;采用免疫组织化学染色检测NLRP3/Caspase-1/GSDMD凋亡基因的表达情况。结果HE染色与肿瘤对照组比较,各治疗组肿瘤生长状态较差,且DOX-3.0、NG/DOX-3.0、DOX-6.0、NG/DOX-6.0肿瘤坏死面积百分率依次增大,组间差异均有统计学意义(P<0.05);与肿瘤对照组比较,DOX-3.0、NG/DOX-3.0、DOX-6.0、NG/DOX-6.0组肿瘤组织中NLRP3、Caspase-1、GSDMD的蛋白阳性表达水平依次减弱,组间差异均有统计学意义(P<0.05),纳米凝胶载药组作用优于单纯阿霉素给药组。结论纳米凝胶载药组治疗H22肝癌的作用优于阿霉素单独用药,它能通过下调NLRP3/Caspase-1/GSDMD通路,有效抑制肿瘤生长、降低药物使用剂量及提高抗癌效果,具有良好发展前景,其作用机制有待于进一步探讨。Objective The mechanism of doxorubicin nanogel loading(NG)on H22hepatomic carcinoma tumorbearing mice was investigated by detecting the NOD-like receptor protein 3(NLRP3)/cysteine aspartate proteolyticase-1(Caspase-1)/quercetin D(GSDMD)pathway.Methods H22hepatoma cell beads were inoculated under the axillary arm of the right forelimb of Balb/c mice×1.25106per bead,and when the tumor volume reached 50~60mm^(3),40 Balb/c mice were randomly divided into 5groups(n=8),which were tumor control group,doxorubicin 3mg group(DOX-3.0),doxorubicin 6mg group(DOX-6.0),and doxorubicin nanogel drug delivery group 3mg group(NG/DOX-3.0)and doxorubicin nanogel loaded 6mg group(NG/DOX-6.0).Each group was treated with tail vein injection,and normal saline(5ml/kg),DOX(3mg/kg),DOX(6mg/kg),NG/DOX(3mg/kg)and NG/DOX(6mg/kg)were given 1 time/3days for 4times,and the animals were sacrificed on the 3rd day after the end of administration.Hematoxylin-eosin(HE)staining was used to observe tumor growth and necrosis area.Immunohistochemical staining was used to detect the expression of NLRP3/Caspase-1/GSDMD apoptosis genes.Results H22hepatoma cell beads were inoculated under the axillary arm of the right forelimb of Balb/c mice×1.25106per bead,and when the tumor volume reached 50~60mm^(3),40Balb/c mice were randomly divided into 5groups(n=8),which were tumor control group,doxorubicin 3 mg group(DOX-3.0),doxorubicin 6mg group(DOX-6.0),and doxorubicin nanogel drug delivery group 3mg group(NG/DOX-3.0)and doxorubicin nanogel loaded 6mg group(NG/DOX-6.0).Each group was treated with tail vein injection,and normal saline(5ml/kg),DOX(3mg/kg),DOX(6mg/kg),NG/DOX(3mg/kg)and NG/DOX(6mg/kg)were given 1time/3days for 4times,and the animals were sacrificed on the 3rd day after the end of administration.Hematoxylin-eosin(HE)staining was used to observe tumor growth and necrosis area.Immunohistochemical staining was used to detect the expression of NLRP3/Caspase-1/GSDMD apoptosis genes.Conclusion The effect of nanogel drug loading group in the treatment of H22

关 键 词：阿霉素 纳米凝胶 免疫组织化学 NLRP3/Caspase-1/GSDMD通路 H22荷瘤 

分 类 号：R73[医药卫生—肿瘤]