运用血清药理学方法观察补肾调冲方对大鼠卵巢颗粒细胞增殖、激素分泌功能以及抑制素B（INHB）、胰岛素样生长因子-1（IGF-1）mRNA表达的影响。方法 向培养的SD大鼠卵巢颗粒细胞中分别加入10%空白血清（对照组）、人转基因重组促卵泡激素（hFSH）含药血清（阳性对照组）、补肾调冲方含药血清（补肾调冲方组），培养48 h，以闪烁计数器测定3H-TdR掺入DNA的量，流式细胞仪测定细胞周期分布，放射免疫法测定培养液中雌二醇（E2）和孕激素（P）水平，并采用RT-PCR法测定INHB、IGF-1 mRNA的表达量。结果 补肾调冲方含药血清可剂量相关地促进3H-TdR掺入颗粒细胞DNA的量。与对照组相比，补肾调冲方各含药血清组G0/G1期细胞均明显减少，而S期细胞则明显增多；E2、P的量明显升高；颗粒细胞INHB、IGF-1 mRNA表达量均明显增多，且呈剂量相关性。结论 补肾调冲方含药血清能明显促进卵巢颗粒细胞的增殖、增强甾体激素的分泌功能，该作用可能通过增强INHB、IGF-1基因的表达，从内分泌和自分泌两条途径调节卵巢功能而实现的。

To observe the effect of serum containing Bushen Tiaochong Recipe (BTR) on rats ovarian granulosa cell proliferation, hormone secretion and inhibin B (INHB) and insulin-like growth factor-1 (IGF-1) mRNA expressions by the method of serum pharmacology. Methods Blank sserum (10%), serum containing follicle-stimulating hormone (FSH) or increasing doses of BTR were added to 6-weeks-old Sprague Dawley rats to prepare the control, positive control, and serum containing BTR groups, in which granulosa cells were incubated for 48 h. [3H] Thymidine deoxyriboside (3H-TdR) incorporation with DNA assay was performed using a scintillation counter. Cell cycle analysis was carried out by flow cytometer. Levels of estrodial and progestogen were examined by radio-immunity. INHB and IGF-1 mRNA expressions were measured by the method of RT-PCR. Results There was a dose-dependent increase in3H-TdR incorporation in granulosa cells in the serum containing BTR. DNA content assay exhibited BTR serum-treated cells in G0/G1 phase decreased markedly, while those in S phase increased significantly compared with the normal serum group. The levels of E2 and P in BTR serum-treated groups were all significantly higher than those in the normal serum group. It showed that serum containing BTR increased more INHB and IGF-1 mRNA expressions in granulosa cells than those in the normal serum group with a dose-dependent manner. Conclusion Serum containing BTR could stimulate granulosa cells proliferation and steroid hormone secretion through promoting INHB and IGF-1 mRNA expressions in granulosa cells. This indicates serum containing BTR could regulate granulosa cells function through two different pathways of endocrine and autocrine.

江苏省自然科学基金资助项目（BK2005006）