Study On The Mechanism Of Qing Luo Tong Bi Compound Compatibility Reducing The Reproductive Toxicity Of Tripterygium

Objective:To observe the reduction effects of Compound Tripterygium Wilfordii (TW) on male reproductive toxicity and to provide the basis for security applications of Tripterygium.Methods:In vivo studies SD rats were randomly divided into blank control group, TW, Compound TW, TW+Rehmanniae Radix (RR), TW+Caulis Sinomenii(CS),TW+Stiff Silkworm(SS),TW+Panax Notoginseng (PN), Compound-RR,Compound-CS, Compound-SS, Compound-PN.The rats in blank control group were given the same volume of the solvent solution. All drugs were serially administered by intragastric administration for four weeks. The content of T, FSH, LH, INHB in serum was examined by ELISA. Testicles and epididymis were got to weigh. One-side epididymis was used to sperm counting, the rest part was fixed with formaldehyde to make pathological section. In vitro studies culture system of spermatocytes and the Sterol cell was established and different doses of triploids were added at different time to settle down the IC50. Influence of each effective component of the Qingluotongbi formula paired with TP on rate of proliferating inhabitation of spermatocytes and the Sterol cells was praised by MTT.Apoptosis of cells in each group was detected by flow cytometry, content of LDH, ABP, INHB by ELISA, expression of Herc4, Mrto4, Ipo11mRNA by Real time PCR and content of Bax, Bcl2, Caspase3 protein in spermatocytes and ABP, TF protein in Sterol cells detected by Western blot.Results:Vivo studies demonstrated that compared with blank control group, the content of T, FSH, LH, INHB decreased in the TW group after serially administered by intragastric administration for four weeks while there was no change in the compound group. RR and PN could regulate the level of serum hormone changed by TW while no significant change occurred in the CS and SS group. Pathology observation showed severe deformation of spermatogenesis tubules, sperm cells of all stages arranging in disorder, cell degeneration or necrosis being obviously visible and inflammatory cell infiltrationetc in TW group. However, there existed no significant pathological change in the compound group. Vitro studies indicated that TP could lead to apoptosis of spermatogenesis cells and the Sterol cells while CAT and TPNS could protect them. With the help of CAT and TPNS, the content of LDH decreased, Herc4, Mrto4, Ipo11mRNA increased, Bax, Caspase3 protein reduced and Bcl2 increased, ABP, INHB, TF protein in the Sterol cell increased while such change hadn’t been found in the CS and SS group.Conclusion:Vivo and vitro study on male reproductive toxicity showed that the toxicity of TW might affect structure and function of testis to decrease the number of sperm, accelerate apoptosis of spermatogenesis cells and the Sterol cells, and damage secretary function of the Sterol cells by interfering structure and function of the pituitary-gonad axis and level of hormone controlling spermatogenesis to affect sperm production. Compound TW can reduce male reproductive toxicity probably by affecting the hypothalamus-pituitary-gonad axis to regulate secretion of endocrine hormone, maintain sperm production, slowing down apoptosis of spermatogenesis cells and the Sterol cells, and promote secretary function of the Sterol cells. The study also indicated RR and PN could reduce male reproductive toxicity while CS and SS could increase the effect in the compound TW.