弗式不完全佐剂诱导小鼠腹腔淋巴管瘤模型方法的改进

Modification of establishing mouse model of abdominal lymphangiomas by injection of incomplete Freund′s adjuvant

目的改进弗式不完全佐剂诱导小鼠腹腔淋巴管瘤模型的建立方法。方法 20只C57BL/6小鼠中的16只按照小鼠体重随机分为两组,分别采取d1、d15(实验Ⅰ组)和d1、d8、d15(实验Ⅱ组)模式每次腹腔注射弗氏不完全佐剂与PBS以1:1体积比配制的乳悬液0.2ml。首次注射后约3个月,脱颈处死小鼠,观察小鼠腹腔内诱导形成的白色肿瘤样组织的生长部位,用游标卡尺记录大小,随后对其进行病理组织学观察,并通过免疫组化表达淋巴管内皮细胞透明质酸受体1(LYVE-1)来进一步证实为淋巴管瘤。另外4只小鼠分别作为两组的对照组(对照Ⅰ组和对照Ⅱ组)按照与实验组对应的注射模式腹腔注射0.2 ml PBS缓冲液。结果实验Ⅰ组、实验Ⅱ组小鼠腹腔内成瘤率均为100%。实验Ⅰ组每只小鼠腹腔成瘤总体积(227.82±124.87)mm3,明显小于实验Ⅱ组的(365.38±78.74)mm3(P<0.05)。实验Ⅰ组、实验Ⅱ组的肿瘤样组织均经HE染色后光学显微镜下观察,及免疫组化均强阳性的表达LYVE-1,证实为淋巴管瘤。对照Ⅰ组、对照Ⅱ组4只小鼠腹腔均未见任何新生物生成。结论利用C57BL/6小鼠腹腔注射弗氏不完全佐剂,以d1、d8、d15注射模式能够诱导获得更多的小鼠腹腔淋巴管瘤。

Objective To modify the establishment of mouse model of abdominal lymphangiomas by injection of incomplete Freund＇s adjuvant. Methods Sixteen C57BL/6 mice in twenty were divided into two groups according to the weight of mice using completely random measures. Mice were intraperitoneally injected with 0. 2 ml each time of emulsified incomplete Freund′s adjuvant （prepared with epual volume of PBS） twice or three times, with the dl, d15 （experimental group I ） and dl, d8, d15 （experimental group Ⅱ ） regime. All mice were sacrificed about 3 months after the first injection. The site of intraabdominal tumor tissues was observed histopathologiealy and the size of tumor tissues was measured by Vernier scale calipers. The expression of lymphatic endothelial hyaluronan receptor-1 （LYVE-1） was detected by immunohistochemistry. Another four mice were injected with 0.2 ml PBS each time as the control group （control group I and the control group U ） with the corresponding injection regime. Results The abodominal tumor formation rate was 100% in experimental groups of I and II. The total tumor volume of experimental group I was （227.82±124. 87） mma, which was less than （365.38±78. 74） mm3 in experimental group Ⅱ （P〈0.05）. Lymphangioma was confirmed by optical microscopy after HE staining and strongly expressed LYVE-1 by immunohistochemistry. There was no intra-abdominal neoplasma in all mice in the two control groups. Conclusion Intraperitoneal injection of incomplete Freund~ s adjuvant using dl, d8, d15 iniection regime can obtained more intra-ahdominal lymphangiomas in C57BL/6 mice.