[Zuogui Pills improve testicular development of offspring rats exposed to prenatal stress via Cx43].

This study aims to reveal the mechanism of prenatal stress in affecting the testicular development of offspring rats and the intervention effects of Zuogui Pills via connexin 43(Cx43). Forty pregnant SD rats were randomized into a blank control group, a mo-del group, a high-dose(18.9 g·kg~(-1)) Zuogui Pills group, a low-dose(9.45 g·kg~(-1)) Zuogui Pills group, and a vitamin E(1.44 mg·kg~(-1)) group. The other groups except the blank control group was subjected to chronic unpredictable mild stress for the modeling of prenatal stress. The model was evaluated by sucrose preference test, open field test, and enzyme-linked immunosorbent assay(ELISA) of the glucocorticoid level. ELISA was employed to measure the thyroxine 4(T4), testosterone(T), and follicle-stimulating hormone(FSH) levels to assess kidney deficiency. Hematoxylin-eosin(HE) staining was employed to evaluate the status of testicular germ cells. An automatic sperm analyzer was used to measure the sperm quality. Immunofluorescence double staining was employed to detect the expression of Cx43 and follicle-stimulating hormone receptor(FSHR) in the testes of offspring rats. The mRNA and protein levels of Cx43, FSHR, phosphatidylinositol 3-kinase(PI3K), and protein kinase B(Akt) were determined by real-time fluorescence quantitative polymerase chain reaction and Western blot, respectively. Prenatal stress induced testicular development disorders in offspring rats. The HE staining results showed that on the day of birth, the model group had reduced seminiferous tubules in the testes, elevated FSH level in the serum, and lowered Cx43 level in the testicular tissue. Male offspring rats of 60 days old had reduced testicular spermatogenic function, decreased sperm quality, elevated FSH level and lowered T level in the serum, and down-regulated protein and mRNA levels of Cx43, FSHR, PI3K, and Akt in the testicular tissue. Zuogui Pills alleviated the abnormal development and dysfunction of testicles in the offspring rats caused by prenatal stress. In summary, Zuogui Pills may weaken the effects of prenatal stress on testicular development and spermatogenic function of offspring rats by activating the PI3K/Akt pathway to regulate Cx43 expression in the testicular tissue.

Zuogui Wan alleviated maternal kidney-yin deficiency-induced thymic epithelial cell dysfunction in newborn rats through Wnt/ß-catenin signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Kidney-yin deficiency (KYD) during pregnancy is common and associated with possibility of thymus hypoplasia in neonates. Zuogui Wan (ZGW) is a classic traditional medicine to treat KYD. AIM OF STUDY: The Wnt/ß-catenin signaling pathway is essential for thymic epithelial cell (TEC) viability, function and for thymus integrity. We evaluated whether maternal diets with ZGW in KYD rats ameliorates epithelial cell dysfunction in the fetal thymus, and investigated its underlying mechanism in which the Wnt/ß-catenin signaling pathway is involved. MATERIALS AND METHODS: Rats were randomly assigned to four groups (n = 8). Two experimental groups received KYD induction with or without ZGW supplementation. The other 2 vehicle groups were sham operated and administrated with normal saline or ZGW. KYD was established using periodically chronic shaken stimulus and threaten stress. Success of the model induction was evaluated by the general observation, changing of the body weight and plasma thyroxine level. Then, pregnant of vehicle and KYD rats were fed with or without ZGW-supplemented diet throughout the F1 gestation. Postnatal thymi samples were obtained after delivery for histological examination. In vitro, TECs of the newborn rats whose mother suffered KYD were isolated, and cultured using the serum containing ZGW with or without the supplement of Wnt4/ß-catenin pathway inhibitor ICG-001. Cell viability was evaluated by CCK-8 assay. Meanwhile, the thymi tissues and TECs were collected for biochemical analysis. Levels of thymosin ß4 (TMSß4) and thymosin α1 (Tα1) were detected by ELISA assay. The mRNA and protein expression of Wnt4, ß-catenin, and Foxn1 were determined by RT-qPCR and Western blot respectively. RESULTS: In vivo, KYD resulted in significantly increased apoptosis of TECs and atrophy of the thymi, especially in the medullary zone. The morphological changes observed in KYD rats were ameliorated by ZGW treatment. Meanwhile, the decreased TMSß4, Tα1, Wnt4, ß-catenin, and Foxn1 levels in KYD rats were also significantly alleviated by ZGW administration. In vitro, elevated TMSß4 and Tα1 levels accompanied with upregulated Wnt4, ß-catenin, and Foxn1 expressions in the TECs were observed after ZGW intervention, however, which were significantly downregulated by ICG-001 supplement. CONCLUSIONS: Maternal kidney-yin deficiency could result in TEC dysfunction in newborn rats. ZGW was able to improve the growth and development of TEC, potentially by regulating the Wnt/ß-catenin pathway.

[Effect of Qianlean Pill on IL-1ß, IL-10, and TNF-α in Prostate Tissues of Chronic Nonbacterial Prostatitis Rats].

OBJECTIVE: To observe the effect of Qianliean Pill (QP) on inflammatory factors such as IL-1ß, IL-10, and tumor necrosis factor α (TNF-α) in chronic nonbacterial prostatitis (CNP) model rats, and to explore its therapeutic mechanism. METHODS: CNP rat model was established by castration and estradiol benzoate injection. Totally 50 rats were randomly divided into 5 groups, i.e., the model group, the positive medicine group, the high dose QP group, the medium dose QP group, and the low dose QP group, 10 in each group. Besides, 10 normal rats were recruited as a normal control group. Since the 8th day of castration, Pulean Tablet (PT) at 10. 80 g/kg was administered to rats in the positive medicine group by gastrogavage. QP at 11.00, 5.50, and 2.75 g/kg was administered to rats in high, medium, and low dose QP groups by gastrogavage. Distilled water at 2 mL/100 g was administered to rats in the model group and the normal control group by gastrogavage, once daily for 30 successive days. After 30 days of medication all rats were sacrificed and their prostate tissues were extracted. The prostatic index was calculated. Pathological changes of rat prostate were observed under light microscope. Meanwhile, levels of IL-1ß, IL-10, and TNF-α were detected using enzyme linked immunosorbent assay. RESULTS: Compared with the normal control group, the prostate index obviously decreased, levels of IL-1ß, TNF-α, and IL-10 in the prostate tissue significantly increased in the model group (P < 0.01). Compared with the model group, the prostate index obviously decreased in high and medium dose QP groups, and the positive medicine group (P < 0.01); levels of IL-1ß, TNF-α, and IL-10 obviously decreased in each QP group and the positive medicine group (P < 0.01). Compared with the positive medicine group, the TNF-α level decreased more obviously in the high dose QP group (P < 0.05). Compared with the normal control group, inflammatory reactions occurred obviously in rats' prostate of the model group. Compared with the model group, inflammatory reactions were milder in rats' prostate of each QP group and the positive medicine group, and their degrees were improved to some extent. CONCLUSION: QP could treat CNP, which might be achieved by regulating local immune state of the prostate, relieving inflammatory reactions of the prostate, and lowering levels of IL-ß, TNF-α, and IL-10 in the prostate tissue.

Effects of astaxanthin and esterified glucomannan on hematological and serum parameters, and liver pathological changes in broilers fed aflatoxin-B1-contaminated feed.

The effects of astaxanthin (ASTA) and esterified glucomannan (EMG) on hematological and serum parameters, and liver pathological changes in broilers fed on aflatoxin-B1 (AFB1) contaminated diet were investigated. Two hundred and forty 10-day-old broilers were randomly assigned to one of five dietary treatments including: (i) control diet; (ii) AFB1-contaminated diet; (iii) AFB1 + EGM diet; (iv) AFB1 + ASTA diet; and (v) AFB1 + EGM + ASTA diet. At 35 days old, blood and liver tissue samples were collected for analysis. Results indicated that total white blood cell (WBC) number, hemoglobin (Hgb) concentration, hematocrit (Hct) level, serum alanine amino transferase (AST) and γ-glutamyl transferase (GGT) activities, red blood cell (RBC) number, serum globulin (GLB) and urea nitrogen (BUN) concentrations (P < 0.05) were increased by feeding AFB1-contaminated diet. EMG and ASTA alleviated the alteration of RBC, WBC, Hgb and AST caused by AFB1-contaminated diet. Liver superoxide dismutase (SOD) activity was reduced, while myeloperoxidase (MPO) activity was increased by AFB1-contaminated diet (P < 0.05). Both EGM and ASTA restrained the increase of MPO activity (P < 0.05). Degeneration of the liver tissues was found in broilers fed AFB1-contaminated diet. It suggested that feeding 0.4 mg/kg AFB1-contaminated diet resulted in adverse effects on blood parameters and liver morphology. Dietary addition of EGM addition at 5 g/kg diet, ASTA at 10 mg/kg diet and especially their combination showed positive protection effects on alleviating the alteration of feeding AFB1. The results indicated that supplementation of 5 g EGM/kg diet, 10 mg ASTA/kg diet and their combination could partially or greatly alleviate the adverse effects caused by AFB1, with the EGM+ASTA group receiving the most effective treatment.

[Effect of qianlie huichun capsule on microstructure and ultranstructure of prostate glandular tissues in rats].

OBJECTIVE: To investigate the effect of Qianlie Huichun capsule on the microstructure and ultranstructure of prostate glandular tissue in the model rat. METHOD: Hynertophy of prostate model rat was established by injecting testosterone to gelding male rats. After having been fed with Qianlie Huichun capsule for 30 days, the rats were killed and prostate tissues were resected for pathomorphological studies with microscope and electromicroscope, and the diameter of glandular lumer and the height of glandular epithelial cells were measured under the microspcope for different groups of rats. RESULT: In the model groups, the glandular epithelial cells mutiplycated notably, showing stratified and pseudostratified cells that made the glandular lumer cramped. Under the electromicroscope, the glandular epithelial cells became high columnor and the rough endoreticulum extremely expanded. But in treatment groups, the change of the diameter of the glandular lumer and the height of the glandular epithelial cells were less remarkable than those in model groups. So the differerence between the model group and the treatment groups was remarkable (P < 0.01). CONCLUSION: Qianlie Huichun capsule can depress the glandular epithelialceu multiplication of prostate gland in model rats.

[Effect of Qianlie Huichun on prostate tissue VEGF expression in rats].

OBJECTIVE: To investigate the effect of Qianlie Huichun on the expression of vascular endothelial growth factor(VEGF) in prostate tissues and expound its anti-prostatomegaly action in model rats with prostatic hypertrophy induced by injected testosterone. METHODS: A total of 60 male rats were eventy randomized into six groups. All were gelded except the normal control group. After a week, the gelded rats were injected with testosterone(4 mg/kg/d), meanwhile the first group were fed with a small dosage of Qianlie Huichun(0.4 g/kg/d), the second group with a medium dosage(0.8 g/kg/d), the third group with a large dosage(1.6 g/kg/d), and the fourth group injected with estriol(2.5 mg/kg/d), all for a month. The fifth group were model controls, and the sixth the normal controls, both fed with the same amount of pure water for a month. Then all the six groups of rats were killed and their prostate glands were resected for the examination of the expression rate of VEGF by immunohistochemical method. RESULTS: The difference of VEGF expression between Qianlie Huichun groups and the model group was significant(P < 0.01), and so was it between the medium and large dosage middle, large amount of Qianlie Huichun groups and the estriol group(P < 0.01). CONCLUSION: Qianlie Huichun depressed the VEGF expression of the prostate gland in model rats, and the expression rate decreased with the increased amount of the drug, which shows that Qianlie Huichun has a definite therapeutic effect on prostatic hypertrophy by depressed the vascular growth of the vessel in the prostate gland.

[Effects of experimental varicocele on the apoptosis of spermatogenic cells in rats].

OBJECTIVE: To explore the effects of experimental varicocele on the apoptosis of spermatogenic cells in rats. METHODS: Thirty-two Sprague-Dawley(SD) male rats were randomly divided into four groups: pseudo-operation group, 45 d experimental group, 60 d experimental group and 90 d experimental group. Experimental varicocele was created by partial ligation of left renal vein. The apoptosis number of spermatogenic cells was measured by flow cytometry. RESULTS: No marked apoptosis peak was observed in the control group, but it was observed in all the other groups, and the height of the peak increased with the duration of experiment. CONCLUSIONS: Varicocele can induce apoptosis of large numbers of spermatogenic cells, and the decrease of spermatogenic cells. This may be the mechanism of varicocele causing male sterility.

[Effect of qianlie huichun capsular on Fas expression and cell apoptosis of prostate gland tissue in rat].

OBJECTIVE: To investigate the effect of Qianlie Huichun capsular on Fas expression and cell apoptosis of prostate gland tissue in the model rat. METHOD: Hypertrophy of prostate model rat was established by injecting testosterone to gelding male rats. After being treated with Qianlie Huichun ig 30 days, the rats were killed and prostate glands were resected for examination. The Fas expression was examined by immunobistochemical SABC. The cell apoptosis and the peak of cell apoptosis in the prostate gland of the rats were examined by flow cytometry. Compared with model group, the weight of prostage gland tissue in groups treated with Qianlie Huichun capsular was light (P < 0.01). RESULT: Compared with model group, the Fas expression in all treatment groups increase(P < 0.05, P < 0.01). Compared with normal and model groups, cell apoptosis in all treatment groups increase at different level(P < 0.01). CONCLUSION: The Qianlie Huchun capsular increases the Fas expression and cell apoptosis of model rats, and shows a definite treatment effect on the hypertrophy of prostate by promoting the apoptosis of prostate cell.