Page 87 - 《中国药房》2021年07期

柠檬苦素对MFC胃癌荷瘤模型小鼠免疫功能及凋亡相关因子表

达的影响 Δ

熊 伟 ，韩 华，陈华敏，吴晓明，黄光钺（海南医学院第二附属医院胃肠外科，海口 570311）
＊

中图分类号 R965 文献标志码 A 文章编号 1001-0408（2021）07-0845-06
DOI 10.6039/j.issn.1001-0408.2021.07.13

摘 要 目的：研究柠檬苦素对MFC胃癌荷瘤模型小鼠免疫功能及凋亡相关因子表达的影响。方法：将MFC胃癌细胞接种于小
鼠右侧腋下以复制 MFC 胃癌荷瘤模型，造模成功后分为模型组、环磷酰胺组（阳性对照，25 mg/kg）和柠檬苦素高、中、低剂量组
（100、50、25 mg/kg），每组10只。除模型组小鼠灌胃0.5％羧甲基纤维素钠外，其余各组小鼠灌胃相应药物，每天1次，连续14 天。
小鼠给药前及末次给药后均测定体质量，并在末次给药后取脾、胸腺、肿瘤组织以计算脾指数、胸腺指数及抑瘤率；采用流式细胞
+
+
+
术检测小鼠CD4 、CD8 T淋巴细胞百分数和CD4 /CD8 比值；采用酶联免疫吸附法检测小鼠血清中免疫功能相关指标[白细胞介
+
素2（IL-2）、IL-10、干扰素γ（IFN-γ）]的表达水平；采用实时定量聚合酶链式反应法和Western blot法检测小鼠肿瘤组织中凋亡相关
因子[细胞色素C（Cyt-C）、B淋巴细胞瘤2蛋白（Bcl-2）、Bcl-2相关X蛋白（Bax）]的mRNA和蛋白的相对表达水平。结果：除环磷
酰胺组小鼠体质量较模型组显著降低外（P＜0.05），其余各组小鼠体质量的差异均无统计学意义（P＜0.05）。环磷酰胺组和柠檬
苦素高、中、低剂量组小鼠的抑瘤率分别为（58.16±7.07）％、（37.09±4.26）％、（27.30±3.64）％、（15.13±2.95）％。与模型组比较，
+
+
环磷酰胺组小鼠脾指数、胸腺指数、CD4 和CD8 T淋巴细胞百分数、CD4 /CD8 比值、血清中IL-2和IL-10的表达水平、肿瘤组织中
+
+
Bcl-2的mRNA和蛋白的相对表达水平以及柠檬苦素各剂量组的IL-10表达水平、Bcl-2的mRNA和蛋白的相对表达水平均显著降
低（P＜0.05）；环磷酰胺组IFN-γ表达水平、Cyt-C和Bax的mRNA和蛋白的相对表达水平，以及柠檬苦素各剂量组的脾指数（低剂
+
量组除外）和胸腺指数、CD4 和CD8 T淋巴细胞百分数、CD4 /CD8 比值、IL-2和IFN-γ的表达水平、Cyt-C和Bax的mRNA和蛋白
+
+
+
的相对表达水平均显著升高（P＜0.05）。结论：柠檬苦素能抑制MFC胃癌荷瘤模型小鼠肿瘤组织的生长，且毒副作用相对较小；
其作用机制与提高免疫功能和诱导凋亡有关。
关键词 柠檬苦素；MFC胃癌细胞；免疫功能；凋亡；小鼠
Effects of Limonin on Immune Function and Apoptosis-related Factors Expression in MFC Gastric Cancer
Bearing Model Mice
XIONG Wei，HAN Hua，CHEN Huamin，WU Xiaoming，HUANG Guangyue（Dept. of Gastrointestinal Surgery，
the Second Affiliated Hospital of Hainan Medical College，Haikou 570311，China）

ABSTRACT OBJECTIVE：To study the effects of limonin on immune function and apoptosis-related factors expression in MFC
gastric cancer bearing model mice. METHODS：MFC gastric cancer bearing model was established by inoculating MFC gastric
cancer cells into the right armpit of mice. After modeling，model mice were divided into model group，cyclophosphamide group
（positive control，25 mg/kg）and limonin high-dose，medium-dose and low-dose groups（100，50 and 25 mg/kg），with 10 mice in
each group. Other groups were given relevant medicine intragastrically，once a day，for consecutive 14 days，except that model
group was given 0.5％ sodium carboxymethyl cellulose intragastrically. Before administration and after last administration，the body
weight of mice was measured；spleen，thymus and tumor tissue were taken after the last administration to calculate the spleen
index，thymus index and tumor inhibition rate. The percentage of CD4 and CD8 T lymphocytes，CD4 /CD8 ratio were detected
+
+
+
+
by flow cytometry. The expression of immune function related indexes（IL-2，IL-10，IFN-γ）in serum were detected by ELISA.
RT-PCR and Western blot assay were adopted to detect relative mRNA and protein expression of apoptosis-related
factors [cytochrome C（Cyt-C），Bcl-2，Bax] in tumor tissue of mice. RESULTS：There was no significant difference in body
weight among the other groups except that of cyclophosphamide group was decreased significantly，compared with model group
（P＜0.05）. Inhibitory rate of tumor were （58.16 ± 7.07）％ ，（37.09 ± 4.26）％ ，（27.30 ± 3.64）％ ，（15.13 ± 2.95）％ in
cyclophosphamide group，limonin high-dose，medium-dose and low-dose groups. Compared with model group，spleen index，
thymus index，the percentages of CD4 and CD8 T lymphocyte cells，CD4 /CD8 ratio，serum levels of IL-2 and IL-10，relative
+
+
+
+
mRNA and protein expression of Bcl-2 in tumor of mice in cyclophosphamide group as well as the expression of IL-10，relative
mRNA and protein expression of Bcl-2 in limonin groups were decreased significantly（P＜0.05）. The expression of IFN-γ，relative
mRNA and protein expression of Cyt-C and Bax of cyclophosphamide group as well as spleen index（except for low-dose group），
Δ 基金项目：海南省卫生健康行业科研项目（No.20A200078） thymus index， the percentage of CD4 + and CD8 + T
+
＊副主任医师。研究方向：胃肠肿瘤的诊断与治疗。电话： lymphocytes，CD4 /CD8 + ratio，the expression of IL-2 and
0898-66809173。E-mail：xw36725747@163.com IFN-γ ，and relative mRNA and protein expression of Cyt-C

中国药房 2021年第32卷第7期 China Pharmacy 2021 Vol. 32 No. 7 ·845 ·