摘要
目的研究和评价云南丹参复合有效部位(DS-MEF)的抗艾滋病病毒1型(human immunodefi-ciency virus type1,HIV-1)活性。方法采用放射性核素3H掺入法、荧光方法和酶联免疫吸附实验分别测定其对HIV-1逆转录酶、蛋白酶和整合酶的抑制作用;采用传代人T淋巴细胞MT-4细胞、慢性HIV-1ⅢB感染传代人T淋巴细胞H9细胞和健康人新鲜外周血单核细胞(peripheral blood mononuclear cell,PBMC),用HIV-1实验室传代的ⅢB病毒株和临床分离的齐多夫定(zidovudine,AZT)敏感株018a及AZT耐药株018c感染细胞测定其对细胞的毒性和在细胞培养内对HIV-1P24的抑制作用;灌胃或腹腔注射1次不同剂量DS-MEF后测定其对昆明种小鼠的急性毒性。结果DS-MEF抑制HIV-1整合酶、逆转录酶和蛋白酶的半数抑制浓度(50%inhibiting concentration,IC50)分别为(2.59±0.50)mg/L、(27.39±11.18)mg/L和(9.38±2.45)mg/L;对MT-4细胞毒性的半数有毒浓度(50%toxic concentration,TC50)为(13.19±6.07)mg/L,抗HIV-1活性的IC50为(0.224±0.163)mg/L,选择指数(selected index,SI)为58.7;在HIV-1ⅢB慢性感染的H9细胞培养内对细胞毒性的TC50为(18.11±9.84)mg/L,抑制HIV-1P24抗原的IC50为(17.230±21.114)mg/L,SI为1.1。对PBMC细胞毒性的TC50为(288.70±0.08)mg/L,抑制AZT敏感株HIV-1018a的IC50为(26.42±11.16)mg/L,SI为10.9,抑制AZT耐药株HIV-1018c的IC50为(27.87±5.35)mg/L,SI为10.4;与AZT和奈韦拉平均有协同抗HIV-1的作用,联合指数分别为0.78和0.67。灌胃20g/kg DS-MEF对小鼠无明显毒性,无毒剂量>20g/kg,腹腔注射后对小鼠的半数致死剂量为1.18g/kg。结论DS-MEF毒性小,具有多环节多功能地抑制HIV-1的作用。
Objective To explore and evaluate the activities of composite extract from Salvia Yunnanensis and in cell cultures （DS-MEF） for inhibition of human immuno-deficiency virus type 1 （HIV-1） in vitro and in cell cultures. Methods The inhibitory activity of DS-MEF on HIV-1 reverse transcriptase （RT）, protease （PR） and integrase （IN） were detected in vitro with radionuclide 3H incorporation, fluorescence assay and enzyme-linked im- munosorbent assay respectively. The human T-lymphocyte MT- 4 cell line, human T-lymphocyte H 9 cell line chronically infected with HIV-1 Ⅲ B, and the fresh peripheral blood mononuclear cell （ PBMC ） of healthy persons as well as the laboratory passed HIV-1 ⅢB and the clinically isolated HIV-1 AZT sensitive 018a or resistant 018c infected cell cultures were used for evaluating the cytotoxicities and inhibitory activities of DS-MEF on HIV-1 P 24 antigen. The acute toxicities of DS-MEF on KM mice were determined by gastric gavages and intraperitoneal injec- tions with various dosages. Results The IC50 of DS-MEF for inhibiting HIV-1 IN, RT and PR were 2.59 ± 0. 50 mg/L, 27.39 ± 11.18 mg/L and 9.38± 2.45 mg/L respectively. In MT-4 cell cultures infected with HIV-1 11], TCs0 were 13. 19±6.07 rag/L, ICso and SI of anti-HIV-1 activity were 0. 224 ±0. 163 mg/L and 58.7; in chroni- cally infected H 9 cell cultures, TCs0 were 18. 11 ±9.84 rag/L, IC50 on HIV-1 P 24 antigen and SI werel7. 230± 21. 114 mg/L and 1.1 respectively; TCs0 in HIV-1 infected PBMC cultures were 288.70 ± 0.08 rag/L; IC50 on AZT sensitive HIV-1 018a： 26.42± 11.16 mg/L, and SI： 10. 9; On AZT resistant HIV-1 018c, ICs0： 27. 87± 5.35 mg/L, and SI： 10. 4. Moreover, DS-MEF showed synergistic effect with AZT or nevirapine （NVP） on HIV- 1 m B in MT-4 cell cultures, the respective combination index was 0.78 or 0. 67. DS-MEF showed no acute toxicity in KM mice with the dosage up to 20 g/kg via gastrogavage, and the 50% lethal dose （ LDs0 ） via intraperitoneal injection was 1.18 g/kg. Conclusion DS-MEF is a promising anti- HIV-1 agent with low toxicity in mice and pos- sesses multi-targets and effective activities.
出处
《中国中西医结合杂志》
CAS
CSCD
北大核心
2008年第8期711-715,共5页
Chinese Journal of Integrated Traditional and Western Medicine
基金
863计划项目"丹参抗艾滋病病毒复合有效部位的临床前研究"(No.2004AA2Z3342)
关键词
丹参
有效部位
抗艾滋病病毒1型活性
协同作用
Salvia Yunnanensis
composite extract
anti-human immuno-deficiency virus type 1 activities
synergistic effect
