皖南蝮蛇抑瘤组分Ⅰ急性毒理研究

Acute toxicity of the agkistrodon halys venom antitumor component-Ⅰ from snake venom in mice

目的 探究皖南蝮蛇抑瘤组分Ⅰ(Agkistrodon Halys venom antitumor component-I,AHVAC-Ⅰ)对小鼠的急性毒理反应,评价AHVAC-Ⅰ作为一种新的抗肿瘤药物使用的安全性。方法 采用小鼠半数致死量法,将120只SPF小鼠随机分为对照组,AHVAC-Ⅰ剂量组(1.00 mg/kg、1.44 mg/kg、、2.96 mg/kg、4.25 mg/kg),每组20只小鼠。按分组给相应剂量腹腔注射给药一次。于第7天,眼球取血检测小鼠肝肾功能(ALT、AST用于检测肝功能;CR、BUN用于检测肾功能),HE染色法检测脏器组织病理形态表现;根据LD_(50)值,1.00 mg/kg与1.44 mg/kg蛇毒组例入低剂量组,2.06 mg/kg蛇毒组为中剂量组,2.96 mg/kg蛇毒组例入高剂量组。结果 小鼠腹腔注射AHVAC-Ⅰ的LD_(50)是2.585 mg/kg,其95%可信限为2.233~3.069 mg/kg。与对照组相比,中、高剂量组小鼠血清中ALT、AST、CR、BUN水平显著升高(P<0.05),而低剂量组小鼠血清中的ALT、AST、CR和BUN均无明显变化。病理检查发现中、高剂量组小鼠的肝、肾有变性损伤。中剂量组肝组织可见部分肝细胞轻度水肿,胞体增大,胞浆透亮;高剂量组肝组织可见肝细胞点状坏死,肝细胞水肿,双核增多;中剂量组肾组织可见部分肾小球水肿,囊腔变小,肾小管轻度水肿;高剂量组肾组织可见肾小球水肿囊腔变小甚至消失,周围肾小管轻度水肿。而低剂量AHVAC-Ⅰ组小鼠肝脏、肾脏、心肌均未发现明显的病理变化。结论 低剂量AHVAC-Ⅰ不会引起小鼠严重的急性毒性反应,当浓度超过2.06 mg/kg对小鼠的主要毒性靶器管为肝脏与肾脏。

Objective To evaluate the safety of agkistrodon halys venom antitumor component-Ⅰ(AHVAC-Ⅰ) from snake venom in clinical use, the acute toxicity test in mice was conducted.Methods 120 experimental mice were divided into blank control group and treatment groups(1.00 mg/kg, 1.44 mg/kg, 2.06 mg/kg, 2.96 mg/kg, and 4.25 mg/kg), with 20 in each group. The modified Karber’s method was used to ascertain the median lethal dose(LD_(50)) and 95% confidence interval. After a single intraperitoneal injection, observation was conducted for 7 days. After 7 days, blood was taken from the eyeball to detect the liver and kidney function of mice in different dose groups(ALT and AST were used to detect liver function;CR and BUN were used to detect renal function), and HE staining was used to detect the histopathological morphological manifestations of main organs. According to the LD;value, the 1.00 mg/kg and 1.44 mg/kg snake venom groups were selected as in the low dose experimental group, the 2.06 mg/kg snake venom group was selected as the middle dose experimental group, and the 2.96 mg/kg snake venom group was selected as the high dose experimental group.Results Treatment with AHVAC-Ⅰ caused significantly behavioral changes. The calculated LD_(50) for AHVAC-Ⅰ in mice via intraperitoneal injection was 2.585 mg/kg and the 95% confidence limit was 2.233~3.069 mg/kg. The blood biochemical parameters showed that alanine aminotransfe(ALT), aspartate amino-transferase(AST), blood urea nitrogen(BUN) and creatinine(CR) were increased significantly in medium(2.06 mg/kg) and high dose(2.96 mg/kg) experimental group compared with the control group(P<0.05). However, no significant difference in liver and kidney function between the control and low groups(1.00 mg/kg, 1.44 mg/kg)(P>0.05). The histopathology showed degeneration in the liver and kidney of the medium(2.06 mg/kg) and high dose(2.96 mg/kg) experimental group. In the medium dose experimental group, some hepatocytes were slightly edematous with enlarged cell bodies and clear cytoplasm. In the high dose experimental group, liver tissue showed punctate necrosis of hepatocytes, hepatocyte edema, and increased binucleation. In the medium dose experimental group, partial glomerular edema was seen in renal tissue, the cyst cavity became smaller, and the renal tubules were slightly edema. In the high dose experimental group, the glomerular edema and cysts were reduced or even disappeared, and the surrounding renal tubules were slightly edema. While, no histopathology changes in low groups(1.00 mg/kg, 1.44 mg/kg).Conclusion Liver might be the target organ after treated with medium and high dose AHVAC-Ⅰ, but low dose AHVAC-Ⅰ has no obvious acute toxicity to mice.