Effects Of Serum Contained BuShentiaoChong On The Secretion Function And Structure Of Ovarian Granulosa Cells From Female Rat In Vitro

Objective: To investigate the effect of BuShenTiaoChong decoction on function and structure in cultured rat ovarian granulosa cells, which in the purpose to explain the mechanism that the decoction is to promote follicular development. Method: PMSG treated femal S.D. rats were used as a model and ovarian granulosa cells were prepared. Rat ovarian granulosa cells were cultured in DMEM/F12 medium containing 2% calf serum. Two groups were set up: study group with rat serum contained BuShenTiaoChong decoction in different dosage and control group with nomol serum. Granulosa cells were cultured in the medium containing 10% different rat serum at 37℃. After 24 hours, the concentration of estradiol (E₂) , progesterone (P) , cAMP, insulin-like growth factor-I (IGF-I) and tumor necrosis factor-a (TNF-α) in culture medium were measured by radioimmunoassay (RIA) ; the ultrastructures changes of granulosa cells were observed under electronicmicroscope. After 12 hours, Percentage of apoptosis and death, bcl-2 and P⁵³ gene protein expression of granulosa cells were quantitatively assessed by flow cytometry (FCM) analysis. Result: RIA illustrated: compared with control group, the serum with medicine could increase E₂, IGF-I production (P<0.05) , and could dcrease P, TNF-a production from GC in vitro (P<0.05) . However, the serum with medicine could not effect on cAMP production from GC (P>0.05) .FCM showed: the percentage of apoptosis of GC in study group was less than that in control group (P<0.05 ) ; and the percentage of death of GC in study group was same as that in control group (P>0.05 ) . The positive expression rate of bcl-2 protein of GC in study group was higher than that in control group (P<0.05 ) ; and the positive expression rate of P⁵³ protein of GC in study group was lower than that in control group (P<0.01) .The pathology study illustrated the main ultrastructureal appearences of granulosa cells in study group were abundance with enlarged smooth endoplasmic reticulum , mitochondria and lipid droplets in cytoplasm.