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Objective To explore the mechanism of ginsenoside Rg1 in alleviating heat stress-induced testicular injury in mice. Methods A total of 20 C57BL/6 male mice (6~8 weeks old) were randomly divided into 4 groups (n=5). The mice from the control group and heat stress (HS) group were intraperitoneally injected with 10 mL/(kg·d) 0.9 % normal saline for 14 d, while those in the HS+Rg1 group and the Rg1 group were given an intraperitoneal injection of 20 mg/(kg·d) for 14 d, and then on the 7th day after administration, the mice in the HS group and the HS+Rg1 group had the lower abdomen put into a 43 ℃ water bath for 30 min as a single heat stress after being anesthetized with 4% chloral hydrate. Mouse spermatocytes GC-2spd(ts) were divided into control group (routine culture for 48 h), HS group (placed in a 43 ℃ water bath for 30 min after 36 h of conventional culture, and cultured till the end of 48 h), HS+Rg1 group (50 μmol/L Rg1 treatment followed by heat stress injury), and Rg1 group (no heat stress injury). In 1 d after modeling, the eyeball blood samples were collected to detect serum testosterone with ELISA, and the testicles were extracted to observe the morphology and weighed to calculate the testicular index. HE staining was used to observe the histopathology of testis, and corresponding reagents and kits was employed to detect the content of malondialdehyde (MDA) and activities of catalase (CAT) and superoxide dismutase (SOD) in testis tissue. After the epididymal sperm were collected, the sperm concentration and motility were analyzed by computer-assisted sperm analysis (CASA) system. In in vitro experiments, cell apoptosis was detected with TUNEL staining, the protein levels of Nrf2, Keap1, HO-1, Bax, Bcl-2 and Caspase3 were detected with Western blotting, and the mRNA levels of GCLC, GCLM and NQO1 were detected by RT-qPCR. Results Rg1 prevented the decreases in testicular weight and testicular index caused by heat stress, reduced the damage of testicular tissue structure, prevented the decrease of sperm concentration and vitality, antagonized the decreasd number of Leydig cells and serum testosterone level, reduced the accumulation of MDA in testicular tissue, and enhanced the activities of CAT and SOD. Rg1 treatment alleviated the apoptosis of GC-2spd(ts) cells, down-regulated the expression of Bax, Caspase3 and Keap1 proteins, enhanced the expression of Bcl-2, Nrf2 and HO-1 proteins, and increased the transcriptional levels of Nrf2 target genes GCLC, GCLM and NQO1. Conclusion Rg1 has no significant effect on the structure and function of mouse testes, but it can effectively improve the ability of mouse testes to resist heat stress injury, which may be related to the activation of Nrf2 signaling pathway, the improvement of antioxidant enzyme activity, and the reduction of apoptosis of spermatogenic cells.

Objectives: High reactive oxygen species (ROS) levels lead to cell death, and the testes are among the most vulnerable organs to oxidative damage. Rg1, an active ingredient extracted from the natural medicine ginseng, has potential anti-inflammatory, antioxidant and antiapoptotic properties. Our previous studies showed that Rg1 can effectively improve spermatogenic function in mice, but the specific mechanism remains unclear. The purpose of this study was to investigate the effect of Rg1 on oxidative stress and spermatogonium apoptosis in D-gal-induced testicular toxicity and elucidate the associated mechanism. Methods: Male C57BL/6 mice at 6–8 weeks of age were intraperitoneally injected with D-gal (200 mg/kg) for 42 days to establish a testicular injury model, and on day 16, 40 mg/kg Rg1-rich saline was injected intraperitoneally. Concurrently, we established an in vitro model of D-gal-damaged spermatogonia, which was treated with Rg1. Results: We found that treatment with the ginsenoside Rg1 reduced D-gal-induced oxidative stress and spermatogonium apoptosis in vivo and in vitro. Mechanistically, we found that Rg1 activated Akt/bad signaling and reduced D-gal-induced spermatogonium apoptosis. Discussion: We provide evidence showing that the antioxidant effect of Rg1 is mediated by the Akt/GSK-3β/NRF2 axis. Based on these findings, we consider Rg1 a potential treatment for testicular oxidative damage. [ABSTRACT FROM AUTHOR]

5-Fluorouracil (5-FU)-injured stromal cells may cause chronic bone marrow suppression; however, the underlying mechanism remains unclear. Angelica sinensis polysaccharide (ASP), the main biologically active ingredient of the Chinese herb, Angelica sinensis (Oliv.) Diels (Apiaceae), may enrich the blood and promote antioxidation. This study investigated the protective antioxidative effects of ASP on perivascular mesenchymal progenitors (PMPs) and their interactions with hematopoietic cells. PMPs were dissociated from C57BL/6 mouse femur and tibia and were subsequently divided into the control, ASP (0.1 g/L), 5-FU (0.025 g/L), and 5-FU + ASP (pre-treatment with 0.1 g/L ASP for 6 h, together with 0.025 g/L 5-FU) then cultured for 48 h. Hematopoietic cells were co-cultured on these feeder layers for 24 h. Cell proliferation, senescence, apoptosis, and oxidative indices were detected, along with stromal osteogenic and adipogenic differentiation potentials. Intercellular and intracellular signaling was analyzed by real-time quantitative reverse transcription polymerase chain reaction and Western blotting. ASP ameliorated the reactive oxygen species production/scavenge balance in PMPs; improved osteogenic differentiation; increased SCF, CXCL12, VLA-4/VCAM-1, ICAM-1/LFA1, and TPO/MPL, Ang-1/Tie-2 gene expression. Further, the ASP-treated feeder layer alleviated hematopoietic cells senescence (from 21.9 ± 1.47 to 12.1 ± 1.13); decreased P53, P21, p-GSK-3β, β-catenin and cyclin-D1 protein expression, and increased glycogen synthase kinase (GSK)-3β protein expression in co-cultured hematopoietic cells. ASP delayed oxidative stress-induced premature senescence of 5-FU-treated feeder co-cultured hematopoietic cells via down-regulation of overactivated Wnt/β-catenin signaling. These findings provide a new strategy for alleviating myelosuppressive stress. [ABSTRACT FROM AUTHOR]