Garg S, Talwar G P, Upadhyay S N
National Institute of Immunology, New Delhi, India.
J Ethnopharmacol. 1998 Apr;60(3):235-46. doi: 10.1016/s0378-8741(97)00157-8.
A novel approach for immunocontraception by intervention of local cell mediated immunity in the reproductive system by using single intrauterine application of neem oil has been described earlier. The reversible block in fertility was reported to last for 107-180 days in female Wistar rats (Upadhyay et al., 1990. Antifertility effects of neem oil by single intrauterine administration: A novel method of contraception. Proceedings Of The Royal Society Of London B 242, 175-180) and 7-11 months in monkeys (Upadhyay et al., 1994. Long term contraceptive effects of intrauterine neem treatment (IUNT) in bonnet monkeys: An alternative to intrauterine contraceptive devices. Contraception 49, 161-167). The present study, describes the identification and characterization of the biologically active fraction from neem seeds (Azadirachta indica A. Juss. Family Meliaceae), responsible for the above activity in adult female Wistar rats. Initial studies with the mechanically extracted oil and solvent extracts of neem seeds have revealed that the antifertility activity was present in constituents of low to intermediate polarity. A hexane extract of neem seeds was reported to be biologically active (Garg et al., 1994. Comparison of extraction procedures on the immunocontraceptive activity of neem seed extracts. Journal of Ethnopharmacology 22, 87-92). Subsequently, hexane extract was sequentially fractionated through the last active fraction using various separation techniques and tested for antifertility activity at each step. Preparative HPLC was used for isolating individual components of the active fraction in quantities, sufficient for characterization. An analytical HPLC method was developed for standardization of the fraction. The active fraction was identified to be a mixture of six components, which comprises of saturated, mono and di-unsaturated free fatty acids and their methyl esters. Dose response study was performed with the last active fractions. The antifertility activity with the active fraction was reversible in nature and it was completely active until 5% concentration. There was no systemic toxic effect following the administration of the active fraction. This study, for the first time, proposes an active fraction from neem seeds, responsible for long term and reversible blocking of fertility after a single intrauterine administration with high efficacy.
此前曾描述过一种新颖的免疫避孕方法，即通过在生殖系统中局部细胞介导免疫的干预，利用印楝油单次宫内给药来实现。据报道，在雌性Wistar大鼠中，生育能力的可逆性阻断持续107 - 180天（Upadhyay等人，1990年。单次宫内给药印楝油的抗生育作用：一种新型避孕方法。《伦敦皇家学会学报B》242卷，175 - 180页），在猴子中持续7 - 11个月（Upadhyay等人，1994年。帽猴宫内印楝治疗（IUNT）的长期避孕效果：宫内节育器的替代方法。《避孕》49卷，161 - 167页）。本研究描述了印楝种子（楝科印楝属植物印楝）中负责上述成年雌性Wistar大鼠活性的生物活性成分的鉴定和表征。对机械提取的印楝油和印楝种子的溶剂提取物的初步研究表明，抗生育活性存在于低至中等极性的成分中。据报道，印楝种子的己烷提取物具有生物活性（Garg等人，1994年。印楝种子提取物提取方法对免疫避孕活性的比较。《民族药理学杂志》22卷，87 - 92页）。随后，使用各种分离技术对己烷提取物进行连续分级分离，直至得到最后一个活性级分，并在每个步骤测试其抗生育活性。制备型高效液相色谱用于大量分离活性级分的各个成分，以进行表征。开发了一种分析型高效液相色谱方法用于该级分的标准化。活性级分被鉴定为六种成分的混合物，包括饱和、单不饱和和二不饱和游离脂肪酸及其甲酯。对最后一个活性级分进行了剂量反应研究。活性级分的抗生育活性本质上是可逆的，在浓度达到5%之前完全有活性。给药活性级分后没有全身毒性作用。本研究首次提出了一种来自印楝种子的活性级分，负责单次宫内给药后长期且可逆地高效阻断生育能力。
