Male infertility induced by heat stress has been attracting more and more attention. Heat stress not only causes apoptosis of spermatocytes but also has adverse effects on Sertoli cells, further damaging spermatogenesis. Lycium barbarum polysaccharide (LBP) is the main bioactive component of Lycium barbarum, which has a protective effect on male reproduction, but its mechanism is still unclear. In this study, our results proved that LBP blocked the inhibitory effect on the proliferation activity of Sertoli cells after heat stress, reversed the dedifferentiation of Sertoli cells induced by heat stress, and ameliorated the structural integrity of the blood-testis barrier. In addition, it increased the expression of the androgen receptor and activated Akt signaling pathway to resist heat-stress-induced injury of Sertoli cells.
Pulmonary hypertension (PH) is a chronic and fatal disease, for which new therapeutic drugs and approaches are needed urgently. Baicalein and baicalin, the active compounds of the traditional Chinese medicine, Scutellaria baicalensis Georgi, exhibit a wide range of pharmacological activities. Numerous studies involving in vitro and in vivo models of PH have revealed that the treatment with baicalin and baicalein may be effective. This review summarizes the potential mechanisms driving the beneficial effects of baicalin and baicalein treatment on PH, including anti-inflammatory response, inhibition of pulmonary smooth muscle cell proliferation and endothelial-to-mesenchymal transformation, stabilization of the extracellular matrix, and mitigation of oxidative stress. The pharmacokinetics of these compounds have also been reviewed. The therapeutic potential of baicalin and baicalein warrants their continued study as natural treatments for PH.
Sertoli cells (SCs) provide physical support and material supply for germ cells and participate in the formation of blood-testis barrier. The number of SCs is directly proportional to the number of germ cells. And mature SCs ensure the growth of germ cells and the production of sperm. In this study, we explored the effect and underlying mechanism of Lycium barbarum polysaccharides (LBP) on primary SCs in young rats. Primary SCs were isolated from the testis of 20-day old rats. The cells were then treated with different concentrations of LBP. Immunocytochemistry was used to detect the expression of Ki67 and the androgen receptor (AR), and western blotting was used to detect the expression of cytokeratin-18 (CK-18), AR and phosphorylated Akt (Ser473) in SCs. The number of SCs increased significantly after LBP treatment, and the 100 mg/mL. LBP group had 14% more cells than the control group. The expression of Ki67 in LBP treated groups also increased significantly. LBP inhibited the expression of cytokeratin 18 in SCs. Besides, LBP increased the expression of AR on SCs and promoted the activation of Akt at the ser473 phosphorylation site. LBP promotes the proliferation of immature SCs in young rats and also accelerates their differentiation and maturation. This seems to be associated with activation of the Akt signaling pathway via up-regulation of AR.
The blood–testis barrier (BTB) of Sertoli cells (SCs) is an important biological barrier that maintains spermatogenesis and provides a favourable microenvironment for spermatogenesis. However, heat stress can directly damage the BTB structural proteins of testicular SCs, leading to dyszoospermia. Wuzi Yanzong Pills (WYP) is a traditional Chinese medicine formula used to treat male reproductive diseases. However, whether WYP could ameliorate heat stress injury in primary SCs extracted from rat testes and BTB proteins remains unknown. Here, treatment with WYP (low, medium and high dose) increased the SC viability and the proliferation of cell antigen Ki67 significantly. Additionally, it promoted SC maturation, which presented in the form of increased androgen receptors (ARs) and decreased cytokeratin 18 (CK-18) in three WYP dose groups. WYP upregulated BTB proteins such as zonula occludens 1 (ZO-1) and occludin across all WYP groups and decreased phosphorylated Akt (p-Akt) in the middle and high-dose groups; however, ZO-1 and occludin recovery were reduced with the presence of Akt inhibitor in WYP groups. WYP improved SC viability and proliferation, and ameliorated dedifferentiation and BTB-proteins damaged by heat stress via Akt signalling. The findings present theoretical support for the effects of WYP in the management of dyszoospermia and male infertility.
Abstract Background: Semen cuscutae is one of the common drugs used in treating male infertility (MI) in Traditional Chinese Medicine (TCM), and semen cuscutae flavonoid (SCF) is the main active component of semen cuscutae. The mechanism of SCF on sertoli cells (SCs) and blood-testis barrier (BTB) in male infertility under heat stress have not been clarified yet. Methods: Here, network pharmacology were used to explore the targets and signaling pathways of SCF in the treatment of MI. Then, in vitro experiments were integrated to verify the results of network pharmacology. In the in vitro experiments, the cell viability was measured by performing the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, Ki67 and AR expression were detected by performing immunohistochemistry staining, and AR, Akt, CK-18, and Occludin levels were measured using western blotting. The Akt inhibitor assay was further performed to elucidate the signaling pathway underlying the function of SCF. Result: The data showed a total of 136 potential therapeutic targets of SCF, including HPS90AA1, SRC, JUN, TP53, MAPK1, AKT1, etc., were identified. In vitro experiments suggested that SCF significantly improved the viability of SCs, upregulated the expression of AR, Akt, Occludin, and Ki67, and downregulated the expression of CK-18. After adding the Akt inhibitor to block the PI3K-Akt signaling pathway, the effect of SCF decreased, indicating that SCF regulates the PI3K-Akt signaling pathway via AR, and then regulates occludin, Ki67, and CK-18. Gene ontology (GO) enrichment and Kyoto Encyclopedia of genes and genomes (KEGG) pathway analyses also suggested that SCF regulates cell proliferation and differentiation mainly via the PI3K-Akt signaling pathway. Conclusions: We suggested that the active components of SCF can promote AR expression, and regulate the PI3K-Akt signaling pathway and its downstream factors (occludin, Ki67, and CK-18), to regulate the viability, proliferation, differentiation, and BTB permeability of SCs, ultimately protecting reproductive ability.
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