Protective effect of Fructus Lycii polysaccharides against time and hyperthermia-induced damage in cultured seminiferous epithelium Yanrong Wang a , Hui Zhao b , Xinsheng Sheng a , Patricia E. Gambino c , Brian Costello c , Krzysztof Bojanowski b, * a Department of Histology and Embryology, Ningxia Medical College, Yinchuan 750004, People’s Republic of China b Sunny BioDiscovery, Inc., 675 Hegenberger Rd., 2nd Floor, Oakland, CA 94621, USA c The Collaborative Group, Ltd., 50 East Loop Rd., Stony Brook, NY 11790, USA Received 1 January 2002; received in revised form 1 June 2002; accepted 20 June 2002 Abstract Lycium barbarum L. (Solanaceae) is a Chinese medicinal plant whose fruits (Fructus Lycii ) are used by Chinese physicians for treatment of infertility. However, the active ingredients and the mechanism of action underlying Lycium ’s fertility-facilitating effects remain unknown. Here we report that Fructus Lycii polysaccharides (FLPS) inhibit time- and hyperthermia-induced structural damage in murine seminiferous epithelium, in vitro. Moreover, we found that FLPS delayed apoptosis in this system, both at normothermic and hyperthermic culture conditions. Oxidative stress was reported to be a major cause of structural degradation and apoptosis in hyperthermic testes, and thus the protective effect of FLPS could implicate an antioxidant mechanism of action. To test this hypothesis we assayed the effect of FLPS on ultraviolet light-induced lipid peroxidation, and cytochrome c reduction by free radicals. We found that FLPS is a potent inhibitor of both of these reactions. Together, these results demonstrate the protective effect of FLPS on time- and hyperthermia-induced testicular degeneration in vitro, indicate the potential mechanism of action for this protective effect, and provide a scientific basis for the traditional use of this plant. # 2002 Elsevier Science Ireland Ltd. All rights reserved. Keywords: Seminiferous tubules; Free radicals; Apoptosis; Tissue morphology; Traditional chinese medicine; Polysaccharides 1. Introduction Fructus Lycii (FL) is a medicinal fruit derived from Lycium barbarum L. (Solanaceae), valued in China for a large variety of beneficial effects. The pro-fertility effect of FL has been first described in the XVI century by the great Chinese herbalist Li Shizhen, and FL has been included in most fertility-promoting Chinese herbal remedies ever since. More recently, a Chinese clinical study performed on a small cohort of 42 patients with low sperm count and/or low sperm motility found that 79% of the group ingesting 15 g/day of FL for 4 months had a significant improvement in the quality of sperm (Yin and Guo, 1993). As it is often the case with traditional herbal medicines, the chemical nature of FL’s active ingredients and their fertility-promoting mechanism of action remain unknown. In order to elucidate these questions, we prepared a highly purified polysaccharide extract from FL and tested it for its protective effects on the testicular tissue in an in vitro model. We used time and temperature as variables to induce structural degeneration of this tissue, which was cultured in the absence of follicle stimulating hormone (FSH) and chorionic gonadotropin (hCG). In this model system, the structure of seminiferous tubules undergoes rapid degeneration during the first 72 h, and this degeneration is greatly enhanced by hyperthermic (37 8C) conditions. The physiologic relevance of the temperature variable is difficult to overstate. In vivo, testicular thermoregulation is a complex process con- trolled by numerous local mechanisms that attempt to maintain testes at conditions ideal for spermatogenesis. Hyperthermia caused by congenital defect (such as cryptorchidism) or disease (varicocele) damages the * Corresponding author. Tel.: 510-235-8839; fax: 510-553-0676 E-mail address: kbojanowski@sunnybiodiscovery.com (K. Bojanowski). Journal of Ethnopharmacology 82 (2002) 169  /175 www.elsevier.com/locate/jethpharm 0378-8741/02/$ - see front matter # 2002 Elsevier Science Ireland Ltd. All rights reserved. PII:S0378-8741(02)00169-1