金复康口服液精简方对顺铂所致肾小管上皮细胞凋亡的作用  

Effect of simplified formula of Jinfukang Oral Liquid on apoptosis of renal tubular epithelial cells induced by cisplatin

作　　者：汪文芳 叶亮 潘铱昕 李羽禅 李仕鑫 杨冰 封亮 贾晓斌 WANG Wen-fang;YE Liang;PAN Yi-xin;LI Yu-chan;LI Shi-xin;YANG Bing;FENG Liang;JIA Xiao-bin(School of Traditional Chinese Pharmacy,China Pharmaceutical University,Nanjing 211198,China)

机构地区：[1]中国药科大学中药学院,江苏南京211198

出　　处：《中国中药杂志》2024年第19期5315-5326,共12页China Journal of Chinese Materia Medica

基　　金：中国药科大学双一流新药品种项目(CPU2022PZQ14);中国药科大学双一流创新团队项目(CPU2018GY11);江苏省六大人才高峰创新团队项目(SWYY-CXTD-004)。

摘　　要：为研究金复康口服液精简方(ALG-12)对顺铂(cisplatin,DDP)所致肾小管损伤的作用,将48只C57小鼠随机分为对照组(control)、模型组(model)、DDP组及DDP联用ALG-12低、中、高剂量组,在构建小鼠皮下Lewis肺癌异位移植瘤模型后连续给药16 d,对小鼠肾组织病理变化、血清肌酐(serum creatinine,Scr)、血尿素氮(blood urea nitrogen,BUN)、肾损伤分子1(kidney injury molecule 1,Kim-1)、中性粒细胞明胶酶相关脂质运载蛋白(neutrophil gelatinase-associated lipocalin,NGAL)、肾组织丙二醛(malondialdehyde,MDA)、总超氧歧化酶(total superoxide dismutase,T-SOD)及肾小管细胞凋亡程度进行分析,考察ALG-12对DDP治疗非小细胞肺癌(non-small cell lung cancer,NSCLC)所致肾损伤的影响。体外构建人肾皮质近曲小管上皮细胞(HK-2)模型,通过cell counting kit(CCK)-8、细胞周期及凋亡实验评价ALG-12含药血清对DDP的HK-2的细胞毒性影响。实时荧光定量PCR(real-time quantitative reverse transcription PCR,RT-qPCR)、蛋白免疫印迹(Western blot)及免疫组化用于分析ALG-12对肿瘤蛋白p53(tumor protein p53,p53)介导的凋亡相关基因的表达水平。结果显示,ALG-12可显著缓解Scr、BUN、Kim-1、NGAL、MDA、T-SOD的异常,减轻肾小管损伤、肾间质纤维化及肾小管细胞凋亡程度,提示ALG-12在体内可以减轻DDP所致的肾损伤。体外细胞实验发现ALG-12含药血清可以抑制DDP造成的HK-2细胞凋亡及周期抑制。另外ALG-12可抑制p53信号通路上共济失调毛细血管扩张突变基因Rad3相关激酶(ataxia-telangiectasia mutated-and Rad3-related gene,ATR)、肿瘤蛋白p53基因(tumor protein p53 gene,Tp53)、Bcl-2绑定组件3(Bcl-2 binding component 3,BBC3)、Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)的转录,降低p53、Bax、剪切的半胱天冬蛋白酶3(cleaved caspase-3)的蛋白表达水平,增加B细胞淋巴瘤/白血病-2(B-cell lymphoma/leukemia 2,Bcl-2)及半胱天冬蛋白酶3(caspase-3)的蛋白表�In order to study the effect of the simplified formula of Jinfukang Oral Liquid(ALG-12)on renal tubular injury induced by cisplatin(DDP),48 C57 mice were divided into control group,model group,DDP group,and DDP combined with low,medium,and high dose groups of ALG-12.The mice were administered for 16 days after the establishment of the subcutaneous Lewis lung cancer heterotopic transplant tumor model of mice.The pathological changes,serum creatinine(Scr),blood urea nitrogen(BUN),kidney injury molecule 1(Kim-1),neutrophil gelatinase-associated lipocalin(NGAL),malondialdehyde(MDA),and total superoxide dismutase(T-SOD)in renal tissue and the degree of renal tubular cell apoptosis were analyzed to investigate the effect of ALG-12 on renal injury induced by DDP treatment on non-small cell lung cancer(NSCLC).The human renal cortex proximal tubule epithelial cell(HK-2)model was constructed in vitro,and the effect of ALG-12 drug-containing serum on HK-2 cytotoxicity of DDP was evaluated by CCK-8,cell cycle,and apoptosis tests.Real-time quantitative reverse transcription PCR(RT-qPCR),Western blot,and immunohistochemistry were used to analyze the expression levels of tumor protein p53-mediated apoptosis-related genes by ALG-12.The results showed that ALG-12 could significantly reduce the abnormalities of biochemical indexes like Scr,BUN,Kim-1,NGAL,MDA,and T-SOD and decrease the degree of renal tubular injury,renal interstitial fibrosis,and renal cell apoptosis,suggesting that ALG-12 could reduce the renal injury induced by DDP in vivo.In vitro cell assay found that ALG-12 containing serum could inhibit apoptosis and cell cycle arrest induced by DDP in HK-2 cells.In addition,ALG-12 inhibited the transcription of ataxia-telangiectasia mutated-and Rad3-related gene(ATR),tumor protein p53 gene(Tp53),Bcl-2 binding component 3 gene(BBC3),and Bcl-2 associated X protein gene(Bax)in the p53 signaling pathway,decreased the protein expression levels of p53,Bax,and cleaved caspase-3,and increased the protein expression levels of B-cell

关 键 词：金复康口服液精简方 顺铂 肾小管损伤 非小细胞肺癌 凋亡 

分 类 号：R285.5[医药卫生—中药学]