目的：研究非病毒基因载体聚酰胺-胺型树枝状大分子（PAMAM）-透明质酸（HA）聚合物的体外细胞毒性。方法采用MTT法和流式细胞术，分别检测合成的多种PAMAM-HA聚合物对HeLa细胞、Bel-7402细胞和HepG2细胞的细胞毒性及其诱导细胞凋亡的能力。结果 PAMAM-HA聚合物的细胞毒性随浓度（50~800 mg·L-1）增加、作用时间（24~72 h）延长、PAMAM的代数（G4，G5）增加及HA的相对分子质量（3850和17200）和接枝密度（5%~25%）降低而增加。与聚合物PAMAM G4-HA3850-5%和PAMAM G5-HA3850-5%孵育24 h，肝癌细胞Bel-7402细胞凋亡率分别为4.5%和9.9%。聚合物与DNA形成的复合物PAMAM G4-HA3850-5%/DNA和PAMAM G5-HA3850-5%/DNA与细胞共孵育24 h后，细胞存活率均在80%以上，较PAMAM G4/DNA和PAMAM G5/DNA毒性降低（P<0.05）。结论经不同接枝量和接枝密度HA修饰的PAMAM细胞毒性降低，是一种较有前途的非病毒基因载体。%OBJECTIVE To study the cytotoxicity of polyamide amine dendrimers (PAMAM)-hyaluronic acid(HA)as a non-viral gene delivery vector in vitro. METHODS PAMAM-HA was synthesized by our laboratory. Cytotoxicities of various polymers on HeLa cells,Bel-7402 cells and HepG2 cells were evaluated by MTT assay. Apoptotic rates were determined by flow cytometry. RESULTS The cytotoxicity of PAMAM-HA polymer increased with the concentration of polymer(50-800 mg · L-1),the time of action(24-72 h), the number of generations of PAMAM(G4,G5) and decrease in the molecular mass(3850,17 200)and the graft density(5%-25%)of HA. After incubation with PAMAM G4-HA3850-5%or PAMAM G5-HA3850-5%for 24 h,the apoptotic rate of hepatoma cells—Bel-7402 cells was 4.5%and 9.9%,respectively. After incubation with complexes of PAMAM G4-HA3850-5%/DNA or PAMAM G5-HA3850-5%/DNA for 24 h,the viability of HeLa cells was more than 80%,which was lower than that of PAMAM G4/DNA和PAMAM G5/DNA. CONCLUSION Cytotoxicity of PAMAM modified by HA of different grafting density and quantity can be reduced ,suggesting the PAMAM is a promising non-viral gene vector.
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