Investigation On Methods Of Male Reproductive Toxicity Induced By Chinese Traditional Medicine

OBJECTIVE: To investigate methods of male reproductive toxicity induced by Chinesetraditional medicine.METHOD: In vitro tests: Spermatogenic cells and sertoli cells from SD rat testis wereisolated and cultured. The effects of Triptolide (TL) and Wilforine (WR), activecomponents extracted from Gufengning which possesses male reproductive toxicityclinically, at different concentrations on cell proliferation were examined using MTTmethod.The expression of pan-cadherin and fshr in the cultured cells was examined usingimmunocytochemistry.In vivo test: SD rats were given daily gavage doses of Gufengning for 3-months. Testes slideswere treated with HE, PAS staining and immunohistochemical technology to examinemale reproductive toxicity.RESULT:In vitro tests: The lowest toxic concentration of TL was 10 μg/L in mix culturedspermatogenic cells and sertoli cells, its IC50 was 1.22 mg/L; the lowest toxicconcentration of TL was 10³ μg/L in cultured sertoli cells, its IC₅₀ was 28.15 mg/L; andthe inhibition rates were related to doses. The lowest toxic concentration of WR was 10⁵μg/L in cultured sertoli cells, its IC₅₀ was 139.26 mg/L. WR showed no toxicity in mixcultured cells.Pan-cadherin expression increased in mix cultured cells at TL concentration levels of 0.1,1 and 10 μg/L. No statistical significant difference in pan-cadherin and fshr expressionwas observed in cultured sertoli cells between TL treated groups and vehicle controlgroup. No statistical significant difference in pan-cadherin and fshr expression wasobserved in mix cultured cells and cultured sertoli cells between WR treated groups andvehicle control group.In vivo tests: Decreased quantity or disappearance of spermatogenic cells, multinucleargiant cells, increasing proportion of stage â…  ～ â…¢ seminiferoustubuli cycle wereobserved in SD rats treated with Gufengning for 3 months. No statistical significantdifference in pan-cadherin expression was observed in testes of control and dosinggroups after termination of dosing. Fshr expression was increased in testes of high dosegroup after termination of dosing. Pan-cadherin expression was increased in mid dose group,but decreased in high dose group after recovery period. No statistical significant differencein fshr expression was observed in testes of control and dosing groups after recovery period.