Change Of Spermatogenic Epithelium Under Condition Of Undifferential Inflammation In Rat Testes

Objective: The present study has investigated the change of testicular spermatogenic epithelium and interstitial tissue with stimulated by a bacterial lipoplysaccharide (LPS) and Indian ink in vivo by using the methods of tissue hemotixylin and eosin (H＆E) staining, tissue immunohistochemistry staining, in situ hybrydization technique, magnetic enzyme immunoassay of serum testosterone and luteinizing hormone (LH).We observed mainly the change of proliferating cell nuclear antigen(PCNA),α-catenin in the spermatogenic epithelium and the production of androgen binding protein (ABP) mRNA in Sertoli cell. Meanwhile, we also assayed the role of LPS and Indian ink on the production of testosterone by testicular Leydig cells,LH by pituitary and testicular macrophage expression of iNOS in LPS or Indian ink-treated rats. According to these, we expect to understand the testicular damaged mechanism in pathological conditions and discuss the effect factors of maitenance and disorder of testicular spermatogenic microenvironment in order to provide reliable reference for diagnosing and treating immune infertility.Methods:Experiments were performed on healthy adult male Wistar rats, weighting about 200-250g.The rats were divided into three groups: control group(D),expriment group 1(S1),expriment group 2(S2). The S1 rats were injected ip with saline containing 1mg LPS/㎏, once every two day, for ten days according to the improved way from O'Bryan report. The other experiment group (S2) rats were induced by intravascular injection of Indian Ink, once a day, for ten days. And the rats in the control group were treated with pyrogen-free saline instead of LPS or Indian Ink. After operation rats were anesthetized with ether, then abdomen were exposed via a midline incision and a sample of blood was collected via inferior vena cava puncture in order to examine serum LH and testosterone. Testes were removed and put in Bourin's fixative, after the testes tissue was fixationed, embeded and sectioned as usual methods of light microscope and immunohistochemistry were involved in order to examine the expession of PCNA,α-catenin,iNOS in testes. The testicular sample used in in situ hybrydization technique was obtained after testicular artery douche with Bourin's fixative containing 1/1000 DEPC.Results: 1 HE staining: In normal control group testicular seminiferous tubules distributing were uniform, seminiferous epithelial was organized well. From the basement membrane to the lumen of the seminiferous tubule there were four or five concentric layers of cells including spermatogonium,primary spermatocyte,second spermatocyte,spermatid and spermatozoon. Testicular interstitial tissue was not damaged, it contains Leydig cells which had a large, sphercal nucleus. Nevertheless, observed under light microscope, the testes of two experiment groups showed that in some seminiferous tubules the seminiferous epithelium became thin and the spermatogenic cells were selectively falling from seminiferous epithelium into the lumine of the tubule, at some region of tubules, the cells appeared to have reversed arranges. Meanwhile, in some othe tubules pathological changes were serious, the spermatogenic cells vanished completely from the seminiferous epithelium or vacuoles and coenocytes were visited in the seminiferous epithelium. In the interstitial tissue the number of cells increased obviously. 2 serum LH and testosterone: 1) serum testosterone: In normal control group serum testosterone concentration was 3.576±1.124ng/ml. After LPS treatment, steroidogenesis was promoted and serum testosterone level increased, it was 5.766±1.141ng/ml. Compared with that of D group it was significant (P<0.05). While Indian Ink had no effect on serum testosterone (P>.05), and it was 4.670±1.060ng/ml. 2) LH: After induced by intravascular injection of Indian Ink, serum LH declined and it was 0.690±0.134mIu/ml. While in control group it was 0.988±0.123mIu/ml (P<0.05). However, compared with that of the control group after determination...