摘要
目的观察丹参酮IIA(TSH)对顺铂(DDP)治疗肺癌裸鼠的抑癌作用、不良反应及免疫功能的影响,探讨TSH影响DDP抑癌作用的机制。方法体外培养NCI-H292人肺癌细胞,建立人肺癌裸鼠皮下移植瘤模型。将60只肺癌裸鼠随机分为模型组、DDP组、DDP+TSH高剂量组、DDP+TSH中剂量组、DDP+TSH低剂量组,每组12只,另取12只裸鼠作为空白对照组,共6组。分别予0.9%氯化钠溶液、DDP溶液、DDP+各浓度TSH处理21 d。记录各组裸鼠体重及肿瘤体积变化,取瘤块称重,计算抑瘤率、肿瘤指数;计算胸腺系数、脾脏系数、肾脏系数;测24 h尿液N-乙酰-β-D-氨基葡萄糖苷酶(NAG)、蛋白量(Upro);测血常规、血肌酐(Scr)、血尿素氮(BUN)、胱抑素C(Cys C)水平;流式细胞仪测定裸鼠T淋巴细胞亚群水平;采用酶联免疫法检测裸鼠血清白介素2(IL-2)和IL-10水平,免疫组化法检测TLR4蛋白表达水平。结果与模型组比较,DDP组、DDP+TSH各剂量组裸鼠抑癌率、体重、尿液Upro/24 h、NAG和血清Scr、BUN、Cys C水平明显升高,T淋巴细胞亚群CD3^+、、CD4^+、CD8^+、CD4^+/CD8^+和NK细胞明显增加,裸鼠瘤体体积和WBC、RBC、PLT计数以及血清IL-2、IL-10水平显著降低(P<0.05);与DDP组比较,DDP+TSH各剂量组裸鼠抑癌率、体重和WBC、RBC、PLT计数明显升高,T淋巴细胞亚群CD3^+、CD4^+、CD8^+、CD4^+/CD8^+和NK细胞显著增加,尿液Upro/24 h、NAG和血清Scr、BUN、Cys C水平以及血清IL-2、IL-10水平显著降低(P<0.05)。免疫组化结果显示DDP+TSH各剂量组TLR4蛋白表达均下调。结论TSH可促进DDP的抑癌作用,并减轻DDP引起的肾毒性和骨髓抑制,提高肺癌裸鼠的T淋巴细胞亚群水平,其作用机制可能与下调IL-2、IL-10的表达和调控TLR4信号通路有关。
Objective To observe the effects of tanshinone IIA(TSH)on the anticancer action and adverse reactions of cisplatin(DDP)in the treatment of lung cancer in nude mice,as well as on the immune function of nude mice.To explore the mechanism of TSH on the anticancer action of DDP.Methods NCI-H292 human lung cancer cells were cultured in vitro,and subcutaneous transplanted tumor model of human lung cancer in nude mice was established.Sixty nude mice with lung cancer were randomly divided into model group,DDP group,DDP+TSH high-dose group,DDP+TSH medium-dose group,and DDP+TSH low-dose group,with 12 mice in each group.They were treated respectively with 0.9%sodium chloride solution,DDP solution,and DDP+different concentrations of TSH for 21 d.Another 12 nude mice were used as blank control group.Record the changes of body weight,tumor volume,tumor blocks in each group.Calculate the tumor inhibition rate and tumor index,thymus coefficient,spleen coefficient and kidney coefficient.Measure the levels of urinary N-acetyl-β-D-glucosaminidase(NAG)and urine protein(Upro)at 24 h,serum creatinine(Scr),blood urea nitrogen(BUN),serum cystatin C(Cys C),and blood routine examination.Flow cytometry was used to determine T lymphocyte subsets in nude mice.Enzyme-linked immunoassay was applied to detect the serum interleukin-2(IL-2)and IL-10 levels in nude mice,and immunohistochemical assay was used to detect the protein expression level of Toll-like receptor 4(TLR4).Results Compared with the model group,the tumor inhibition rate,body weight,Upro/24 h and NAG were significantly increased in DDP group and all DDP+TSH dose groups,so were the levels of Scr,BUN and Cys C,and the levels of T lymphocyte subsets CD3+,CD4+,CD8+,NK cells,and the CD4+/CD8+.The tumor volume,WBC,RBC,PLT count and serum IL-2 and IL-10 levels were significantly decreased in DDP group and all DDP+TSH dose groups(P<0.05).Compared with the DDP group,the tumor inhibition rate,body weight,WBC,RBC,PLT count of nude mice in all TSH dose+DDP groups were significantly increased,and the levels of T lymphocyte subsets CD3+,CD4+,CD8+,CD4+/CD8+and NK cells were also significantly increased,but the Upro/24 h,NAG,Scr,BUN,Cys C,IL-2 and IL-10 levels were significantly decreased(P<0.05).Immunohistochemical results showed that the TLR4 protein expression was down-regulated in all DDP+TSH dose groups.Conclusion TSH could strengthen the antitumor effect of DDP,and alleviate the nephrotoxicity and bone marrow suppression induced by DDP in nude mice with lung cancer.It also could improve the immune function of nude mice with DDP treatment.The mechanism may be related to the downregulation of IL-2 and IL-10 levels and TLR4 signal path.
作者
陈煜
凌霄
余丽梅
邱振
席加喜
CHEN Yu;LING Xiao;YU Limei;QIU Zhen;XI Jiaxi(Guangxi Dermatology Hospital,Nanning,Guangxi,530021,China;Guangxi Zhuang Autonomous Region People's Hospital,Nanning,Guangxi,530021,China)
出处
《肿瘤药学》
CAS
2020年第3期287-293,共7页
Anti-Tumor Pharmacy
基金
广西卫健委自筹经费科研课题(Z20180362)。
关键词
丹参酮IIA
顺铂
肺癌
裸鼠
抑癌作用
肾毒性
骨髓抑制
TLR4
Tanshinone IIA
Cisplatin
Nude mice
Lung cancer
Antitumor
Renal toxicity
Bone marrow suppression
TLR4
作者简介
陈煜,女,硕士研究生,研究方向:临床药学;通信作者:席加喜,男,药理学硕士,副主任药师,研究方向:抗肿瘤药物及其不良反应。
