电针预处理联合促性腺激素释放激素激动剂对早发性卵巢功能不全大鼠卵巢储备功能的保护作用OA

Objective:To observe the protective effect of electroacupuncture(EA)pretreatment combined with gonadotropin-releasing hormone agonist(GnRH-a)on ovarian reserve function in a rat model of premature ovarian insufficiency(POI).Methods:Sixty SPF-grade female Sprague-Dawley(SD)rats with regular estrous cycles were randomly assigned,using a random number table,into a blank control group,a model group,an EA group,a GnRH-a group,and an EA combined with GnRH-a group,with 12 rats in each group.Except for the blank group,all rats re-ceived daily intraperitoneal injection of 4-vinylcyclohexene dioxide(VCD)at 80 mg/kg body weight for 15 consecu-tive days to establish the POI model.Beginning 4 weeks before modeling,the EA group and the EA combined with GnRH-a group received EA pretreatment at the bilateral Zhongliao(BL33),Tianshu(ST25),and Guilai(ST29),using a continuous wave with a current of 0.1～1 mA and a frequency of 1～3 Hz,with needles retained for 20 min,5 times per week for 4 weeks.Two weeks before modeling,the GnRH-a group and the EA combined with GnRH-a group received a single subcutaneous injection of GnRH-a at 0.4 μg/rat.The blank group and the model group re-ceived no intervention.When all rats in the model group exhibited estrous cycle disturbance(indicating successful modeling),rats in each group were sacrificed.The ovarian index was measured;serum levels of anti-Müllerian hor-mone(AMH),progesterone(PROG),follicle-stimulating hormone(FSH),luteinizing hormone(LH),and estradiol(E2)were determined by enzyme-linked immunosorbent assay(ELISA);ovarian histomorphology was observed un-der a microscope and follicles at each developmental stage were counted;and granulosa cell apoptosis in ovarian tissue was detected by TUNEL assay,with the proportion of positive cells to total cells(AOD)used as the evaluation in-dex.Results:Compared with the blank group,the model group showed a significant decrease in ovarian index(P＜0.01);serum AMH and PROG levels decreased while serum FSH and LH levels increased(all P＜0.05),with no statistically significant difference in serum E2(P＞0.05);the proportion of preantral follicles in ovarian tissue was significantly reduced(P＜0.01)and that of atretic follicles increased(P＜0.05);and AOD was significantly elevated(P＜0.01).Compared with the model group,the ovarian index increased in the EA group(P＜0.05),whereas no statistically significant difference was observed in the GnRH-a group or the EA combined with GnRH-a group(P＞0.05).In the EA group,serum FSH increased(P＜0.05)and serum LH decreased significantly(P＜0.01),with no statistically significant differences in serum AMH,E2,or PROG(P＞0.05).In the GnRH-a group,serum AMH in-creased significantly(P＜0.05)while serum LH and PROG decreased significantly(P＜0.01),with no statistically significant differences in serum FSH or E2(P＞0.05).In the EA combined with GnRH-a group,serum AMH in-creased(P＜0.05),serum FSH decreased(P＜0.05),and serum LH and PROG decreased significantly(P＜0.01),with no statistically significant difference in serum E2(P＞0.05).The proportion of preantral follicles in ovarian tissue in-creased significantly(P＜0.01)and that of atretic follicles decreased(P＜0.05)in the EA group,GnRH-a group,and EA combined with GnRH-a group.AOD decreased in the EA group(P＜0.05),showed no statistically significant difference in the GnRH-a group,and decreased significantly in the EA combined with GnRH-a group(P＜0.01).Compared with the GnRH-a group,the EA combined with GnRH-a group showed no statistically significant difference in ovarian index(P＞0.05);serum AMH was significantly higher(P＜0.01)and serum FSH significantly lower(P＜0.01),with no statistically significant differences in serum LH,E2,or PROG(P＞0.05);the proportion of primary follicles in ovarian tissue was sig-nificantly lower(P＜0.01);and AOD was significantly lower(P＜0.01).Conclusion:EA,GnRH-a,and EA combined with GnRH-a pretreatment before modeling all exert a certain protective effect on ovarian reserve function in rats with ovarian in-jury.Compared with GnRH-a pretreatment alone,EA combined with GnRH-a pretreatment before modeling exhibits a syn-ergistic effect-upregulating serum AMH,downregulating serum FSH,protecting primary follicles,and reducing granulosa cell apoptosis-thereby better protecting ovarian reserve function in POI rats.