OBJECTIVE: To examine the hormonal regulation of 72-kd type IV collagenase-gelatinase-matrix metalloproteinase-2 in human granulosa-lutein cells. DESIGN: The effect of E2 and GnRH agonist (GnRH-a) on matrix metalloproteinase-2 amount was examined on a long-term granulosa-lutein cell cultures. PATIENTS: The preovulatory follicles from women participating in our IVF program were aspirated through the vaginal wall. Follicular fluids were centrifuged and the granulosa cells were separated and pooled for culturing. MAIN OUTCOME MEASURES: Matrix metalloproteinase-2 enzyme activity was assayed with type IV collagen degradation assay and zymography. The matrix metalloproteinase-2 immunoreactive protein was studied by using specific antibodies. RESULTS: Matrix metalloproteinase-2 was the predominant form of gelatinase in cultured granulosa-lutein cells. The enzyme activity as well as the immunoreactive protein increased in the culture medium of the granulosa-lutein cells in the presence of E2. The effect of E2 was abolished by GnRH-a. CONCLUSIONS: Estradiol regulates the matrix metalloproteinase-2 enzyme amount in granulosa-lutein cells; the effect may be antagonized by GnRH-a.
OBJECTIVE: To examine the hormonal regulation of 72-kd type IV collagenase-gelatinase-matrix metalloproteinase-2 in human granulosa-lutein cells. DESIGN: The effect of E2 and GnRH agonist (GnRH-a) on matrix metalloproteinase-2 amount was examined on a long-term granulosa-lutein cell cultures. PATIENTS: The preovulatory follicles from women participating in our IVF program were aspirated through the vaginal wall. Follicular fluids were centrifuged and the granulosa cells were separated and pooled for culturing. MAIN OUTCOME MEASURES: Matrix metalloproteinase-2 enzyme activity was assayed with type IV collagen degradation assay and zymography. The matrix metalloproteinase-2 immunoreactive protein was studied by using specific antibodies. RESULTS:Matrix metalloproteinase-2 was the predominant form of gelatinase in cultured granulosa-lutein cells. The enzyme activity as well as the immunoreactive protein increased in the culture medium of the granulosa-lutein cells in the presence of E2. The effect of E2 was abolished by GnRH-a. CONCLUSIONS:Estradiol regulates the matrix metalloproteinase-2 enzyme amount in granulosa-lutein cells; the effect may be antagonized by GnRH-a.
Authors: Luciana Andrea Ribeiro; Maria Elena Turba; Augusta Zannoni; Maria Laura Bacci; Monica Forni Journal: BMC Dev Biol Date: 2006-11-30 Impact factor: 1.978