槲皮素自微乳的抗肿瘤作用及其机制分析

目的:研究槲皮素自微乳(quercetin self-nanoemulsifying drug delivery system,Q-SNEDDS)抑制小鼠肉瘤细胞S180的分子机制。方法:制备槲皮素自微乳和空白自微乳,对其粒径和Zeta电位分别进行检测。体内实验分6组:空白组、环磷酰胺(20 mg·kg-1)组、槲皮素自微乳高、中、低剂量(50,25,12.5 mg·kg-1)组、槲皮素组(50 mg·kg-1)。采用肿瘤细胞移植法建立S180移植瘤模型,用药10 d后,检测各组抑瘤率、肝指数、脾指数和胸腺指数等指标;采用实时荧光定量PCR(Realtime PCR)检测各组S180细胞的B细胞淋巴瘤-2(Bcl-2),Bcl-2相关X蛋白(Bax),蛋白激酶B(Akt)mRNA表达的情况;蛋白免疫印迹法(Western blot)检测各组细胞Bax,Bcl-2,Akt,磷酸化-Akt(p-Akt)的蛋白表达。结果:与空白组比较,槲皮素自微乳、环磷酰胺及槲皮素组均能抑制肿瘤细胞生长,槲皮素自微乳组抑瘤率高于槲皮素组(P0.05);环磷酰胺组脏器指数低于空白组(P0.05),槲皮素自微乳组脏器指数高于环磷酰胺组(P0.05)。与空白组比较,槲皮素自微乳上调S180细胞Bax mRNA和蛋白表达(P0.05),下调S180细胞Bcl-2 mRNA和蛋白的表达(P0.05),对Akt蛋白表达影响不明显,但是可以下调Akt mRNA和p-Akt蛋白的表达(P0.05)。结论:槲皮素自微乳具有体内抑制S180细胞的作用,槲皮素自微乳抑瘤作用高于槲皮素组。槲皮素自微乳可能通过抑制Akt信号通路,发挥其抗肿瘤的作用。

Anti-tumor Effect and Mechanisms of Quercetin Self-nanoemulsifying Drug Delivery System

Objective: To investigate the anti-tumor effect of quercetin self-nanoemulsifying drug delivery system (Q-SNEDDS) on mice sarcomarosarcoma cell S180 and explore its molecular mechanism. Method: Q-SNEDDS and Blank-SNEDDS were prepared, and then their particle size and Zeta potential were measured respectively. The in vivo experiment was divided into 6 groups:blank group, cyclophosphamide(CTX) group (20 mg·kg^-1), high dose Q-SNEDDS group (50 mg·kg^-1), middle dose Q-SNEDDS group (25 mg·kg^-1), low dose Q-SNEDDS group (12.5 mg·kg^-1) and quercetin control group (50 mg·kg^-1). S180 bearing mice models were established by using tumor cell transplantation method. After 10 days of drug administration, the inhibition rate and indexes of organ (liver, spleen and thymus) were measured in all groups. B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax), and protein kinase B (Akt) mRNA expression levels of S180 were measured by Real-time quantitative polymerase chain reaction (Real-time PCR). The protein expression levels of Bax, Bcl-2, Akt and p-Akt were measured by Western blot method. Result: Q-SNEDDS, CTX and quercetin groups could inhibit the growth of tumor cells as compared with blank group, and the inhibition rate in Q-SNEDDS groups was higher than that in Quercetin group (P<0.05). The organ indexes of CTX group were lower than those in blank group (P<0.05) and the indexes of Q-SNEDDS group were higher than those in CTX group (P<0.05). As compared with blank group, Q-SNEDDS up-regulated Bax mRNA and protein expression (P<0.05). Q-SNEDDS down-regulated Bcl-2 mRNA and protein expression (P<0.05), down-regulated Akt mRNA and p-Akt protein, but Akt protein expression didn''t change obviously. Conclusion: Q-SNEDDS played the anti-tumor role in vivo, and the effect was superior to quercetin. Its mechanism may be associated with inhibiting Akt signaling pathway.