石见穿多糖对肝腹水H22荷瘤小鼠的作用及可能机制  

Effect of PSSC on tumor bearing mice(H22)in vivo and in vitro and JAK/STAT signaling pathway

作　　者：蔡兴锐 黄芬[1] 刘韵 许丹妮 曾江正[1] CAI Xingrui;HUANG Fen;LIU Yun;XU Danni;ZENG Jiangzheng(Internal Medicine Oncology, The First Affiliated Hospital of Hainan Medical University, Haikou 570102, China)

机构地区：[1]海南医学院第一附属医院肿瘤内科,海南海口570102

出　　处：《西部医学》2022年第7期954-959,965,共7页Medical Journal of West China

基　　金：国家自然科学基金(81960549)。

摘　　要：目的 探讨石见穿多糖(PSSC)对肝腹水H22荷瘤小鼠的作用效果及可能机制。方法 选取BALB/c小鼠80只,建立H22荷瘤小鼠模型,随机分为5组:对照组、环磷酰胺组(CTX,20 mg/kg)、PSSC组(20、40和120 mg/kg),每组16只。观察各组小鼠腹部周长、体重、生存率及生存时间;采用脾淋巴细胞转化试验检测T、B淋巴细胞增殖情况;E-玫瑰花结实验观察T细胞的免疫功能状态;ELISA法检测血清和腹水中白介素2(IL-2)、γ-干扰素(IFN-γ)、肿瘤坏死因子-α(TNF-α)、血管内皮生长因子(VEGF)含量;TUNEL法检测H22细胞凋亡;Caspase比色法测定Caspase-3和Caspase-8酶活性;Western blot法检测Janus激酶1(Jak1)、信号传导和转录激活因子-3(Stat3)、Bcl2-相关X蛋白(Bax),B淋巴细胞瘤-2(Bcl-2)蛋白表达。结果 与对照组相比,PSSC组的腹部周长和体重显著下降,而存活率和存活时间呈剂量依赖性增加;与CTX组相比,PSSC 40、120 mg/kg组腹部周长和体重下降,而存活率和存活时间升高(均P<0.05)。与对照组相比,PSSC组小鼠SI及EtRFC随PSSC治疗剂量的增加而增加;与CTX组相比,PSSC 120 mg/kg组SI及EtRFC百分比升高(均P<0.05)。ELISA检测结果,与对照组相比,PSSC组H22肿瘤小鼠血清和腹水中的IL-2、IFN-γ和TNF-α水平呈剂量依赖性增加,而VEGF表达降低;与CTX组相比,PSSC 120 mg/kg组小鼠血清和腹水中的IL-2、IFN-γ和TNF-α表达升高,而VEGF表达下降(均P<0.05)。TUNEL实验显示,与对照组相比,PSSC呈剂量依赖性增加H22细胞凋亡数量;与对照组相比,随着PSSC浓度的增加,Caspase-3和Caspase-8表达上升(P<0.05);Western blot检测结果,PSSC组Jak1、Stat3和Bcl-2蛋白表达较对照组降低,而Bax/Bcl-2比值上升。结论 石见穿多糖对H22荷瘤小鼠具有一定的抗肿瘤作用,其机制可能与免疫调节与抗凋亡作用有关。Objective To explore the effect and possible mechanism of PSSC on H22 Tumor bearing mice with hepatoma ascites.Methods Eighty BALB/C mice were selected to establish H22 tumor-bearing mice model,and randomly divided into 5 groups:control group,cyclophosphamide group(CTX,20 mg/kg),PSSC group(20,40 and 120 mg/kg),with 16 mice in each group.Abdominal circumference,weight,survival rate and survival time of mice in each group were observed.T and B lymphocyte proliferation were detected by spleen lymphocyte transformation test.T cell immune function was observed by E-rosette experiment.The concentrations of IL-2,γ-interferon,TNF-αand VEGF in serum and ascites were detected by ELISA.Apoptosis of H22 cells was detected by TUNEL assay.The activities of Caspase-3 and Caspase-8 were determined by Caspase-colorimetry.The expressions of Janus kinase 1(Jak1),signaling and transcription activator-3(Stat3),Bcl2-related X protein(Bax)and B lymphocytoma-2(Bcl-2)were detected by Western blot.Results Compared with the control group,the abdominal circumference and body weight of PSSC group decreased significantly,while the survival rate and survival time increased in a dose-dependent manner;Compared with CTX group,PSSC 40 and 120mg/kg groups decreased abdominal circumference and body weight,but increased survival rate and survival time(P<0.05).Compared with the control group,SI and EtRFC in PSSC group increased with the increase of PSSC treatment dose;Compared with CTX group,the percentage of SI and EtRFC in PSSC 120mg/kg group increased(P<0.05).Compared with the control group,the levels of IL-2,IFN-γand TNF-αin serum and ascites of H22 Tumor Mice in PSSC group increased in a dose-dependent manner,while the expression of VEGF decreased.Compared with CTX group,the expressions of IL-2,IFN-γand TNF-αin serum and ascites of mice in PSSC 120 mg/kg group were increased,while the expression of VEGF was decreased.TUNEL showed that PSSC increased the apoptosis of H22 cells in a dose-dependent manner compared with the control group.Compar

关 键 词：石见穿多糖 小鼠 脾淋巴细胞 E-玫瑰花结 肝腹水 

分 类 号：R735.7[医药卫生—肿瘤]