摘要
目的 观察中药复方桔梗白散对Lewis肺癌小鼠皮下移植瘤生长及组织中 B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、胱天蛋白酶(caspase)-3、caspase-9表达的影响.方法 建立Lewis肺癌移植瘤小鼠模型,将70只SPF级雄性C57BL/6小鼠随机分为空白组、模型组、低剂量实验组、中剂量实验组、高剂量实验组、对照组、联合组.空白组、模型组灌胃0.9％NaCl 0.2 mL;对照组给予等量0.9％NaCl灌胃的同时腹腔注射顺铂25 mg·kg-1;高、中、低剂量实验组分别给予193、96、48 mg·kg-1·d-1桔梗白散0.2 mL灌胃;联合组以96 mg·kg-1·d1桔梗白散0.2 mL灌胃,同时腹腔注射顺铂25 mg·kg-1,隔日1次.以流式细胞术检测小鼠骨髓髓源性抑制细胞(MDSCs),以免疫荧光法观察肿瘤细胞中Bcl-2、Bax、caspase-3、caspase-9的表达.结果 空白组、模型组、低剂量实验组、中剂量实验组、高剂量实验组、对照组、联合组小鼠骨髓中MDSCs 百分比分别为(32.50±2.76)％、(63.13±3.14)％、(48.43±2.23)％、(42.53±1.28)％、(32.93±3.56)％(51.30±4.25)％和(19.90±6.21)％;模型组、低剂量实验组、中剂量实验组、高剂量实验组、对照组、联合组Bax荧光强度分别为 10.42±0.68、12.40±1.23、15.14±0.65、22.95±1.76、27.18±1.62 和31.61±1.28,Bcl-2 荧光强度分别为 36.85±0.80、33.92±4.20、28.88±1.01、20.04±2.21、15.69±2.36 和 6.05±0.73,caspase-3 荧光强度分别为5.28±0.44、7.63±0.55、9.66±0.85、14.73±1.18、17.95±1.29 和22.92±1.95,caspase-9 荧光强度分别为 9.48±0.90、11.57±0.72、13.45±0.93、15.73±1.44、19.20±0.96 和 23.21±1.51.中、高剂量实验组的上述指标与模型组比较,在统计学上差异均有统计学意义(均P＜0.05);联合组的上述指标与对照组相比,在统计学上差异均有统计学意义(均P＜0.05).结论 桔梗白散能够通过上调Bax、caspase-3、caspase-9表达,下调Bcl-2表达,抑制MDSCs,促进肿瘤细胞凋亡,抑制肿瘤生长.
Abstract
Objective To observe the effects of traditional Chinese medicine compound Platycodon grandiflorum Bai powder on the growth of subcutaneously implanted tumor and the expression of B-cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(Bax),cysteinyl aspartate specific proteinase(caspase)-3 and caspase-9 in subcutaneously implanted tumor of Lewis lung cancer mice.Methods The model of transplanted tumor of Lewis lung cancer in mice was established.Seventy SPF male C57BL/6 mice were randomly divided into blank group,model group,low dose experimental group,medium dose experimental group,high dose experimental group,control group and combined group.Blank group and model group were given 0.9％NaCl 0.2 mL by gavage;control group was given 0.9％NaCl by gavage and 25 mg·kg-1cisplatin intraperitoneally;high,medium,low dose experimental groups were given 193,96,48 mg·kg-1·d-1 Platycodon grandiflorum Bai powder 0.2 mL by gavage,respectively;combined group was given 96 mg·kg-1·d-1 Platycodon grandiflorum Bai powder 0.2 mL by gavage,and 25 mg·kg-1 cisplatin intraperitoneally,once every other day.The myelogenous suppressor cells(MDSCs)of mouse bone marrow were detected by flow cytometry,and the expressions of Bel-2,Bax,caspase-3 and caspase-9 in tumor cells were detected by immunofluorescence.Results The percentage of MDSCs in bone marrow of mice in blank group,model group,low dose experimental,medium,high dose experimental group,control group and combination group were(32.50±2.76)％,(63.13±3.14)％,(48.43±2.23)％,(42.53±1.28)％,(32.93±3.56)％,(51.30±4.25)％and(19.90±6.21)％,respectively.The fluorescence intensities of Bax in model group,low dose experimental group,medium dose experimental group,high dose experimental group,control group and combination group were 10.42±0.68,12.40±1.23,15.14±0.65,22.95±1.76,27.18±1.62 and 31.61±1.28;Bel-2 were 36.85±0.80,33.92±4.20,28.88±1.01,20.04±2.21,15.69±2.36 and 6.05±0.73;caspase-3 were 5.28±0.44,7.63±0.55,9.66±0.85,14.73±1.18,17.95±1.29 and 22.92±1.95;caspase-9 were 9.48±0.90,11.57±0.72,13.45±0.93,15.73±1.44,19.20±0.96 and 23.21±1.51.There were significant differences between medium,high dose experimental groups and model group(all P＜0.05),and there were significant differences between combined group and control group(all P＜0.05).Conclusion Platycodon grandiflorum Bai powder can up-regulating the expression of Bax,caspase-3 and caspase-9,down-regulating the expression of Bel-2,inhibiting MDSCs,promoting tumor cell apoptosis and inhibiting tumor growth.
基金项目
国家自然科学基金(82160872)
甘肃省教育厅"双一流"科研重点基金(GSSYLXM-05)
甘肃省中医药研究中心开放课题(zyzx-2020-zx17)
中医学一级学科"岐黄英才"导师专项(2022-13)
甘肃中医药大学研究生创新创业训练计划()
国家科技期刊平台
NSTL
万方数据