Celastrol Prevents Oxidative Stress Effects on FSHR, PAPP, and CYP19A1 Gene Expression in Cultured Human Granulosa-Lutein Cells

Abstract

Regulation of oxidative stress (OS) is important to prevent damage to female reproductive physiology. While normal OS levels may have a regulatory role, high OS levels may negatively affect vital processes such as folliculogenesis or embryogenesis. The aim of this work was to study OS induced by glucose, a reactive oxygen species generator, or peroxynitrite, a reactive nitrogen species generator, in cultured human granulosa-lutein (hGL) cells from oocyte donors, analyzing expression of genes involved in oocyte maturation (FSHR, PAPP, and CYP19A1) and OS damage response (ALDH3A2). We also evaluated the effect of celastrol as an antioxidant. Our results showed that although both glucose and peroxynitrite produce OS increments in hGL cells, only peroxynitrite treatment increases ALDH3A2 and PAPP gene expression levels and decreases FSHR gene expression levels. Celastrol pre-treatment prevents this effect of peroxynitrite. Interestingly, when celastrol alone was added, we observed a reduction of the expression of all genes studied, which was independent of both OS inductors. In conclusion, regulation of OS imbalance by antioxidant substances such as celastrol may prevent negative effects of OS in female fertility. In addition to the antioxidant activity, celastrol may well have an independent role on regulation of gene expression in hGL cells.

Celastrol and Melatonin Modify SIRT1, SIRT6 and SIRT7 Gene Expression and Improve the Response of Human Granulosa-Lutein Cells to Oxidative Stress

“…Analysis of gene expression in hGL cells treated with glucose showed no variation in sirtuins’ gene expression ( Figure 3 B, Figure 4 B and Figure 5 B), even though previous studies demonstrated an increased ROS/RNS by the addition of glucose to culture medium [ 38 ]. An increase in DNA damage and in the number of dead cells was observed ( Figure 2 ).…”

“…The addition of OS inductors (glucose or peroxynitrite) to cultured hGL cells increased DNA damage and the percentage of dead cells compared to control, most prominently under peroxynitrite treatment ( Figure 1 A). Previous studies from our laboratory showed that higher ROS/RNS levels in cells treated with glucose or peroxynitrite [ 38 ] increased OS levels, leading to DNA damage and cell death. In this article, we report that the addition of antioxidants (celastrol or melatonin) to culture medium elicits an increase in the amount of high/medium level of 8-OHdG staining (a measure of DNA damage) compared to the amount found in control cells, but in contrast, we found an increase in the total number of surviving cells ( Figure 1 B).…”

“…In this article, we report that the addition of antioxidants (celastrol or melatonin) to culture medium elicits an increase in the amount of high/medium level of 8-OHdG staining (a measure of DNA damage) compared to the amount found in control cells, but in contrast, we found an increase in the total number of surviving cells ( Figure 1 B). In the case of celastrol, this pro-survival effect could be related to the decrease in ROS/RNS level as upregulation of SIRT7 gene expression was observed in cultured cells treated with antioxidant [ 38 ]. The pro-survival response of melatonin is lower and consistent with the fact that the addition of melatonin to cultured medium did not substantially modify ROS/RNS levels ( Figure 2 ).…”

The landscape of aging

“…Accumulating evidence demonstrates that age-related GC dysfunction contributes to decreased oocyte quality. Excessive ROS production also causes the downregulation of FSH-receptor expression in GCs, resulting in an adverse response to FSH and subsequent impaired follicle development with age (Martín-Ramírez et al, 2021). Furthermore, advanced glycation end products (AGEs) lead to protein damage and increases in inflammatory reactions, eventually inducing ovarian function decline (Lliberos et al, 2020;Pertynska-Marczewska and Diamanti-Kandarakis, 2017).…”

Mechanisms of and Potential Medications for Oxidative Stress in Ovarian Granulosa Cells: A Review

“…Table 1 summarizes the mechanisms and effects of some drugs on OS in GCs. Celastrol [160,172] Human granulosa-lutein cells Induces SIRT7 gene expression. Improves cell viability.…”