目的 探讨同仁乌鸡白凤丸对免疫性卵巢早衰模型小鼠的治疗作用及其作用机制。方法 采用皮下多点注射小鼠透明带多肽3建立免疫性卵巢早衰小鼠模型。将小鼠随机分为对照组、模型组、戊酸雌二醇片组、同仁乌鸡白凤丸（1.35、2.70、5.40 g/kg）组，各组连续ig给药8周。测定小鼠血浆雌二醇（E₂）、卵泡刺激素（FSH）、促性腺激素释放激素（GnRH）、抗缪勒管激素（AMH）水平，观察卵巢组织病理学改变并计数各级卵泡数，检测小鼠脾细胞悬液CD3⁺、CD4⁺、CD8⁺细胞数量及比例，比较小鼠卵巢组织B细胞淋巴瘤-2‌（Bcl-2）、Bcl-2相关X蛋白（Bax）、半胱氨酸天冬氨酸蛋白酶-3（Caspase-3）、血管内皮生长因子（VEGF）、碱性成纤维细胞生长因子（bFGF）表达的变化情况。结果 与模型组比较，同仁乌鸡白凤丸1.35 g/kg组小鼠血浆E₂、FSH、GnRH水平明显升高，同仁乌鸡白凤丸2.70、5.40 g/kg组小鼠血浆GnRH水平明显升高（P＜0.05、0.01）。同仁乌鸡白凤丸5.40 g/kg组小鼠血浆AMH水平明显升高（P＜0.05）；同仁乌鸡白凤丸各剂量组原始卵泡数、总卵泡数显著升高（P＜0.01）；同仁乌鸡白凤丸2.70、5.40 g/kg组初级卵泡数、次级卵泡数显著升高，闭锁卵泡数显著降低（P＜0.01）。同仁乌鸡白凤丸1.35 g/kg组小鼠脾细胞悬液中CD8⁺细胞百分比明显升高，CD4⁺/CD8⁺明显降低；同仁乌鸡白凤丸2.70 g/kg组小鼠脾细胞悬液中CD3⁺、CD4⁺、CD8⁺细胞比例均明显升高、CD4⁺/CD8⁺明显降低；同仁乌鸡白凤丸5.40 g/kg组小鼠脾细胞悬液中CD8+细胞比例明显升高（P＜0.05、0.01）。同仁乌鸡白凤丸各剂量组小鼠卵巢Bax蛋白相对表达水平、Bax/Bcl-2均明显降低（P＜0.01）；同仁乌鸡白凤丸1.35、2.70 g/kg组小鼠卵巢Caspase-3蛋白相对表达水平明显降低（P＜0.05）。同仁乌鸡白凤丸2.70、5.40 g/kg组小鼠卵巢VEGF蛋白相对表达水平和同仁乌鸡白凤丸1.35、2.70 g/kg组小鼠卵巢bFGF蛋白相对表达水平均明显增加（P＜0.05、0.01）。结论 同仁乌鸡白凤丸能够调节免疫性卵巢早衰引起的性激素水平紊乱，促进卵巢功能恢复，提高卵巢储备功能，可能与调节机体免疫失衡、抑制卵巢细胞凋亡、调节相关生长因子表达水平等因素相关。

Objective To investigate the effect of Tongren Wuji Baifeng Pills on immune premature ovarian failure model mice and its mechanism. Methods The mouse model of immune premature ovarian failure was established by subcutaneous injection of mouse zona pellucida polypeptide 3. Mice were randomly divided into control group, model group, Estradiol Valerate Tablets group, and Tongren Wuji Baifeng Pills (1.35, 2.70, 5.40 g/kg ) groups, and administered by gavage for 8 weeks. The levels of E₂, FSH, GnRH, and AMH in mice were measured. Pathological changes of ovarian tissues were observed and the number of follicles at all levels was counted. The number and percentage of CD3⁺, CD4⁺, CD8⁺cells in mouse spleen cell suspension were detected. The expression of Bcl-2, Bax, Caspase-3, VEGF, and bFGF in mice ovarian tissues were compared. Results Compared with the model group, the plasma levels of E₂, FSH and GnRH in the Tongren Wuji Baifeng Pills 1.35 g/kg group were significantly increased, while the plasma GnRH levels in the Tongren Wuji Baifeng Pills 2.70 and 5.40 g/kg groups were significantly increased (P < 0.05, 0.01). The plasma AMH level in the Tongren Wuji Baifeng Pills 5.40 g/kg group was significantly increased (P < 0.05), the number of primary follicles and total follicles in the all dose of Tongren Wuji Baifeng Pills group was significantly increased (P < 0.01), the number of primary and secondary follicles and the number of closed follicles in the Tongren Wuji Baifeng Pills 2.70 and 5.40 g/kg groups were significantly increased, while the number of closed follicles was significantly decreased (P < 0.01). The percentage of CD8⁺ cells in the spleen cell suspension of mice in the Tongren Wuji Baifeng Pills 1.35 g/kg group was significantly increased, and the CD4⁺/CD8⁺ ratio was significantly decreased, the percentages of CD3⁺, CD4⁺, and CD8⁺ cells in the spleen cell suspension of mice in the Tongren Wuji Baifeng Pills 2.70 g/kg group were significantly increased, and the CD4⁺/CD8⁺ ratio was significantly decreased, the percentage of CD8⁺ cells in the spleen cell suspension of mice in the Tongren Wuji Baifeng Pills 5.40 g/kg group was significantly increased (P < 0.05, 0.01). The protein relative expression levels of Bax and Bax/Bcl-2 in the ovaries of mice in each dose group of Tongren Wuji Baifeng Pills were significantly decreased (P < 0.01), the protein relative expression levels of Caspase-3 in the ovaries of mice in the Tongren Wuji Baifeng Pills 1.35 and 2.70 g/kg groups were significantly decreased (P < 0.05). The protein relative expression levels of VEGF and bFGF protein in the ovaries of mice in the Tongren Wuji Baifeng Pills 2.70 and 5.40 g/kg groups and the Tongren Wuji Baifeng Pills 1.35 and 2.70 g/kg groups were significantly increased (P < 0.05, 0.01). Conclusion Tongren Wuji Baifeng Pills can regulate the sexual hormone level disorder caused by immune premature ovarian failure, promote the recovery of ovarian function, and improve ovarian reserve function, which may be related to regulating immune imbalance, inhibiting ovarian cell apoptosis, regulating the expression level of related growth factors.

R287.4

北京市科技计划项目（Z241100009024041）