消水散对人胃癌小鼠腹膜移植模型B7/CD28/CTLA-4协同刺激通路及腹水微环境的影响  

Effects of Xiaoshui powder on B7/CD28/CTLA-4 costimulatory pathway and ascites microenvironment in a mouse peritoneal transplantation model of human gastric cancer

作　　者：彭银花 赵晔[1] 彭巍[1] 程晓燕[1] 袁晓清[1] 唐建清[1] 龚辉 PENG Yinhua;ZHAO Ye;PENG Wei;CHENG Xiaoyan;YUAN Xiaoqing;TANG Jianqing;GONG Hui(Department of Oncology,Hunan Academy of Traditional Chinese Medicine Affiliated Hospital,Changsha,Hunan 410013,China)

机构地区：[1]湖南省中医药研究院附属医院肿瘤三科,湖南长沙410013

出　　处：《安徽医药》2022年第4期654-659,共6页Anhui Medical and Pharmaceutical Journal

基　　金：湖南省中医药研究院科研课题(201813)。

摘　　要：目的 研究消水散对人胃癌小鼠腹膜移植模型B7/CD28/细胞毒性T淋巴细胞相关抗原-4(CTLA-4)协同刺激通路及腹水微环境的影响。方法 2019年4月至2020年5月,在湖南省中医药研究院动物中心实验室,采用腹腔注射人胃癌细胞MGC803建立人胃癌小鼠腹膜移植模型,共分为模型组、消水散低、中、高剂量组,另取正常小鼠为对照组,每组10只。通过行为学观察及测定血清促肾上腺皮质激素(ACTH)、皮质醇(CORT)、三碘甲状腺原氨酸(T3)、甲状腺素(T4)、促甲状腺激素(TSH)含量评估模型质量,影像学方法评估恶性腹水情况,流式细胞仪及实时荧光定量PCR(RT-qPCR)分别检测B7-1、B7-2、CD28及CTLA-4表达量及其mRNA表达水平,酶联免疫吸附试验(ELISA)检测腹膜组织转化生长因子-β1(TGF-β1)、白细胞介素-10(IL-10)表达水平。结果 正常小鼠的阳虚症状积分为(0.00±0.00)分,血清ACTH水平为(103.46±10.13)ng/L、CORT水平为(81.34±9.21)μg/L、T3含量为(178.52±15.86)ng/L、T4含量为(213.59±16.97)μg/L和TSH含量为(10.84±1.15)mIU/mL。与对照组相比,模型小鼠阳虚症状积分[(7.84±1.59)分]显著增加(P<0.05),血清ACTH[(71.09±6.42)ng/L]、CORT[(43.42±7.14)μg/L]、T3[(103.77±13.24)ng/L]、T4[(127.64±13.25)μg/L]、TSH含量[(8.14±1.02)mIU/mL]显著降低(P<0.05)。模型组小鼠的腹水量为(5.18±0.46)mL、瘤体积为(1.23±0.12)cm3、瘤重为(1.73±0.15)g、B7-1 mRNA水平为(1.06±0.10)、B7-2 mRNA水平为(1.04±0.11)、CD28 mRNA水平为(0.97±0.08)、CTLA-4 mRNA水平为(1.03±0.09)、TGF-β1含量为[(181.73±15.26)pg/g]和IL-10水平为[(68.74±6.03)pg/g]。与模型组相比,消水散低、中、高剂量组小鼠的腹水量[(4.03±0.41)、(3.24±0.25)、(2.33±0.27)mL]、瘤体积[(0.93±0.09)、(0.49±0.05)、(0.16±0.02)cm3]、瘤重[(1.42±0.13)、(0.79±0.11)、(0.27±0.08)g]、CTLA-4 mRNA水平[(0.83±0.07)、(0.42±0.05)、(0.16±0.02)]、TGF-β1[(152.69±12.39)、(131.45±11.68)、(107.26±10.52)pg/Objective To study the effects of Xiaoshui powder on the B7/CD28/cytotoxic T lymphocyte associated antigen-4(CTLA-4) costimulatory pathway and ascites microenvironment in a mouse peritoneal transplantation model of human gastric cancer.Methods This study was conducted in the Animal Central Laboratory of Hunan Academy of Traditional Chinese Medicine from April 2019 to May 2020. In this study, MGC803 human gastric cancer cells were injected intraperitoneally to establish a mouse peritoneal transplantation model of human gastric cancer. The mice were divided into the model group and the low-dose, medium-dose and high-dose Xiaoshui powder groups, and normal mice were used as the control group, with 10 mice in each group. The model quality was evaluated by behavioral observation and determination of serum adrenocorticotropic hormone(ACTH), cortisol(CORT), triiodothyronine(T3), thyroxine(T4), and thyroid-stimulating hormone(TSH). The condition of malignant ascites was evaluated by imaging. The expression levels of B7-1, B7-2, CD28 and CTLA-4 and their mRNA expression levels were detected by flow cytometry and real-time quantitative PCR(RT-qPCR), respectively. The expression levels of transforming growth factor-β1(TGF-β1) and interleukin-10(IL-10) were detected by enzyme-linked immunosorbent assay(ELISA).Results The score of Yang deficiency symptoms in normal mice was(0.00±0.00)points, the serum ACTH level was(103.46±10.13) ng/L, the CORT level was(81.34±9.21) mg/L, and the T3 content was(178.52±15.86) ng/L, the T4 content was(213.59±16.97) mg/L, and the TSH content was(10.84±1.15) mIU/mL. Compared with the control group, the scores of Yang deficiency symptoms [(7.84±1.59) points] in model mice increased significantly(P < 0.05), and the contents of ACTH [(71.09±6.42) ng/L], CORT [(43.42±7.14) μg/L], T3 [(103.77±13.24) ng/L], T4 [(127.64±13.25) μg/L] and TSH [(8.14±1.02)mIU/mL] in serum were significantly lower(P < 0.05). The ascites volume, tumor volume and tumor weight of mice in the model group were(5.

关 键 词：中草药 胃肿瘤 消水散 人胃癌小鼠腹膜移植模型 B7/CD28/细胞毒性T淋巴细胞相关抗原-4协同刺激通路 腹水 肿瘤微环境 小鼠 近交BALB C 

分 类 号：R285.5[医药卫生—中药学]