Effect Of Aroclor 1254 On Mouse Granulosa Proliferation And Induction To Differentiation In Vitro

Aroclor 1254 is a mixture of environmental hormones that existed extensively in organism circle and did reproductive impairment to human and animals. A mouse granulosa cells culture system in vitro was established and the influence of Arocolor 1254 on mouse granulosa cells'proliferation and differentiation was evaluated in this study. The results were showed as follow:1. Mouse granulosa cells primary culture showed that the group adding EGF (20 ng/mL) and FSH (50 ng/mL), but insulin (2μg/mL), both have significant effect on promoting proliferation (P<0.05). It indicates that EGF and FSH both are factors to promote mouse granulosa cells'proliferation, but insulin is not a promoter.2. The effect of estradiol and Aroclor 1254 on mouse granulose proliferation was compared. The result showed that adding estradiol and adding 0.1 ng / mL Aroclor 1254 had similar effect on mouse granulose cells'proliferation; different concentration of Aroclor 1254 all promoted the mouse granulosa's proliferation significantly(P<0.05), which indicates that Aroclor 1254 has a effect similar to estradiol and can promote mouse granulosa cells'proliferation.3. Immunohistochemical identification showed that LHR was found to express on mouse granulose cells after insulin's stimulation, its P4 amount was lower than control group,but E2 amount was lower, which indicates that insulin promote mouse granulose cells to differentiate. FSH had weaker effect to induce luteinlization than insulin. The medium from insulin and FSH treatment group was collected for radioimmunoassay. That result showed that FSH both promoted mouse granulose cells'E2 and P4 secretion; however, as the time went on, the P4 amount was lower than insulin treatment group.These results indicate that FSH can induce mouse granulosa cells to differentiate in vitro, but not as strong as insulin.4. Adding Aroclor 1254 during mouse granulose cells luteinization process showed that Aroclor 1254 both decreased LHR expression; Aroclor 1254 promoted granulosa cells'estradiol secretion, but decreased luteinized granulosa cells'progesterone secretion detected by hormone radioimmunoassay. It indicates that Aroclor 1254 can block mouse granulosa cells to luteilize in vitro while promote granulosa cells'proliferation. The results above provided scientific evidences for evaluating the reproductive toxical effect of Aroclor 1254 on female individuals .