益气健脾解毒方含药血清对奥沙利铂诱导人肝癌HepG2细胞凋亡的影响

The effect of Yiqi-jianpi-jiedu recipe decoction containing serum on oxaliplatin-induced HepG2 human hepatoma cell

目的探讨益气健脾解毒方含药血清对奥沙利铂诱导人肝癌HepG2细胞凋亡及凋亡抑制基因（Survivin）、caspase-3及caspase．9表达的影响。方法运用血清药理学方法制备大鼠益气健脾解毒方含药血清，将人肝癌HepG2细胞按随机数字表法分为中药组、化疗组、联合组和空白组，空白组加入10％空白血清；化疗组加入2ug奥沙利铂和10％空白血清；联合组加入2ug奥沙利铂和10％益气健脾解毒方含药血清；中药组加入10％益气健脾解毒方含药血清。采用MTT法检测细胞抑制率，采用Westernblot法及RT-PCR法检测各组细胞Survivin、caspase-3、caspase．9蛋白及基因表达。结果与空白组比较，中药组、化疗组及联合组人肝癌HepG2细胞抑制率明显增高（P〈0．05）。与化疗组及中药组比较，联合组Survivin[（0．151±0．054）比（0．288±0．089）、（0．375±0．063）]蛋白表达下调（P〈0．05或P〈0．01），SurvivinmRNA[（0．205±0．091）比（0．487±0．073）、（0．725±0．092）]表达下调（P〈0．05或P〈0．01）；caspase．3[（0．821±0．079）比（0．634±0．098）、（O．487±0．z02）]、caspase．9蛋白[（O．901±0．047）比（0．709±0．054）、（0．402±0．012）]表达上调（P〈0．05或尸〈0．01），caspase．3mRNA[（1．928±0．226）比（1．564±0．195）、（1．287±0．312）]、caspase．9mRNA[（2．063±0．517）比（1．536±0．084）、（1．019±0．182）]表达均上调（P〈0．05或P〈0．01）。结论益气健脾解毒方对奥沙利铂诱导HepG2细胞凋亡机制可能通过增强抑制Survivin及促进Caspase．3及caspase．9蛋白表达发挥作用。

Objective To explore the effect of Yiqi-jianpi-jiedu recipe decoction containing serum on Oxaliplatin-induced HepG2 Human hepatoma cell survivin, caspase-3, caspase-9 expression. Methods The serum pharmacological method was used to prepare nourishing Qi and invigorating spleen decoction containing serum of rat. The HepG2 human hepatoma cells were divided into Chinese herb containing serum group, chemotherapy with Oxaliplatin （OXA） group and the combination group （medicine containing serum and OXA）, and the blank group without drugs. Apply MTT assay, Western Blotting method and RT-PCR method to detect the cell inhibition rate and survivin, caspase-3, caspase-9 protein and gene expression changes. Results Compared with the blank group, the inhibition rate of HepG2 Human hepatoma cell was significantly higher in Chinese herb containing serum group, chemotherapy group and the combination group （P 〈 0.05）. Compared with thechemothcrapy group and Chinese herb containing serum group, the Survivin （0.151 ± 0.054 vs. 0.288 ± 0.089, 0.375 ± 0.063） and the expression of Survivin mRNA （0.205 ± 0.091 vs. 0.487 ± 0.073, 0.725 ± 0.092） of the conbination group downgraded significantly lower （P 〈 0.05 or P〈 0.01）. Compared with chemotherapy group and Chinese herb containing serum group, the caspase-3 （0.821 ± 0.079 vs. 0.634±0.098, 0.487 ± 0.102）, caspase-9 （0.901 ± 0.047 vs. 0.709 ± 0.054, 0.402 ± 0.012） expression, caspase-3 mRNA （1.928 ± 0.226 vs. 1.564 ± 0.195, 1.287 ± 0.312） and caspase-9 mRNA （2.063 ± 0.517 vs. 1.536 ± 0.084, 1.019 ± 0.182） expression of the conbination group upgraded significantly higher （P 〈 0.05 or P 〈 0.01）. Conclusions Nourishing-Qi and invigorating spleen decoction containing serum may affect Oxaliplatin-induced HepG2 apoptosis mechanism through increasing the suppression on Survivin and promoting the expression of caspase-3 and caspase-9 protein and genes. This research provided important experimental basis for clinical application of traditional Chinese medicine treating liver cancer.