口服细胞穿膜肽Penetratin对小鼠肝脏、肾脏和肠道的急性毒性反应研究  

Study of acute toxic reaction of oral cell-penetrating peptide Penetratin in mouse liver,kidney and intestine

作　　者：魏振宇 李林[1] 尉秀清[1] Wei Zhenyu;Li Lin;Wei Xiuqing(Department of Gastroenterology,the Third Affiliated Hospital of Sun Yat-sen University,Guangzhou 510630,China)

机构地区：[1]中山大学附属第三医院消化内科,广州510630

出　　处：《新医学》2020年第6期422-427,共6页Journal of New Medicine

基　　金：国家自然科学基金(81470848);广州市民生科技攻关项目(201903010099)。

摘　　要：目的研究口服细胞穿膜肽Penetratin对小鼠肝脏、肾脏以及肠道的急性毒性反应,评估其安全性。方法将24只Balb/c雄性小鼠随机分为正常对照组、低剂量组和高剂量组3组,每组各8只小鼠。各组小鼠均予正常饮食,其中正常对照组予200μl生理盐水灌胃,低剂量组、高剂量组分别予200μl由生理盐水配制的浓度为5、100μmol/L的Penetratin灌胃。各组小鼠予不同处理24 h后收集标本,采集标本为小鼠的血液、肝脏、肾脏以及肠道组织,用生化法检测各组小鼠血清中ALT、AST、尿素氮、肌酐水平,用ELISA检测血清中二胺氧化酶(DAO)及脂多糖的含量,用实时荧光定量PCR方法检测肝脏、肾脏及肠道炎症因子TNF-α、IL-1β、IL-6以及IL-10的mRNA表达水平,用HE染色评估肝脏、肾脏以及肠道病理损伤情况,TUNEL法检测肝脏、肾脏以及肠道细胞凋亡情况。结果各组小鼠的ALT、AST、尿素氮、肌酐水平、DAO及脂多糖含量、炎症因子TNF-α、IL-1β、IL-6以及IL-10的mRNA表达水平比较差异均无统计学意义(P均>0.05);与正常对照组对比,低剂量组和高剂量组各脏器均未见明显损伤及炎性细胞浸润,细胞凋亡无明显增多。结论口服细胞穿膜肽Penetratin对小鼠的肝脏、肾脏以及肠道均无明显急性毒性反应,作为肠道促吸收剂安全性较好。Objective To observe the acute toxic reaction and evaluate the safety of oral cellpenetrating peptide Penetratin in mouse liver,kidney and intestine.Methods Twenty-four male Balb/c mice were randomly and evenly divided into the normal control,low-dose and high-dose groups.All mice in each group were given a normal diet.In the control group,the animals were gavaged with 200μl of physiological saline.In the low-dose group,the mice were gavaged with 200μl of Penetratin at a concentration of 5μmol/L,and 200μl of 100μmol/L Penetratin was given in the high-dose group.The blood,liver,kidney and intestinal samples were collected 24 h later.The expression levels of ALT,AST,BUN,and Cr in the mouse serum in each group were measured by biochemical methods.The levels of diamine oxidase(DAO)and lipopolysaccharide(LPS)were detected by ELISA.The expression levels of inflammatory cytokines including TNF-α,IL-1β,IL-6 and IL-10 mRNA in each organ were detected by RT-PCR.The pathological injury of liver,kidney and intestinal tissues was evaluated by HE staining.The cellular apoptosis of each organ was detected by TUNEL staining.Results The expression levels of ALT,AST,BUN,Cr,the contents of DAO and LPS,and the expression levels of TNF-α,IL-1β,IL-6 and IL-10 mRNA did not significantly differ among three groups(all P>0.05).Compared with the normal control group,no significant pathological injury or inflammatory cellular infiltration was observed and the cellular apoptosis was not significantly aggravated in the low-dose and high-dose groups.Conclusion Oral cell-penetrating peptide Penetratin yields no evident acute toxicity in mouse liver,kidney and intestine,and it can serve as a safe intestinal absorption enhancer.

关 键 词：细胞穿膜肽 Penetratin 组织损伤 炎症 细胞凋亡 

分 类 号：R965[医药卫生—药理学]