目的 考察乌鸡白凤软胶囊（WJBF）对原发性痛经大鼠的改善作用及可能的作用机制。方法 SPF级雌性SD大鼠随机分为对照组、模型组、布洛芬缓释胶囊(BLF,0.08 g·kg^-1)组、定坤丹(1.4 g·kg^-1)组和WJBF低、中、高剂量(0.3、0.6、1.2 g·kg^-1)组，每组10只。采用im苯甲酸雌二醇联合ip缩宫素制备大鼠原发性痛经模型，第4天起各给药组按剂量ig给药，连续7 d。末次给药1 h后，对照组ip 0.9%氯化钠溶液，其余各组大鼠ip缩宫素，记录扭体潜伏期及30 min内扭体次数；检测子宫与卵巢脏器指数；采用酶联免疫吸附实验（ELISA）检测大鼠血清前列腺素F_2α(PGF_2α)、前列腺素E₂（PGE₂）、血栓素B₂（TXB₂）、6-酮-前列腺素F_1α(6-keto-PGF_1α)、β-内啡肽（β-EP）、5-羟色胺（5-HT）、雌二醇（E₂）、孕酮（Pg）含量；ELISA或生化试剂盒检测子宫组织中PGF_2α、PGE₂、环氧化酶2（COX-2）、Ca²⁺含量。离体子宫平滑肌实验：SPF级雌性大鼠连续2 d sc苯甲酸雌二醇2.0 mg·kg^-1，于第3天麻醉后迅速剖取子宫制备1 cm子宫环，将其悬挂于离体灌流装置中，通入95%O₂，施加前负荷2.0 g，用多导生理记录仪分别记录WJBF(0.37、0.74、1.48 mg·mL^-1)对离体子宫平滑肌自发收缩及乙酰胆碱（Ach）、缩宫素诱导强烈收缩的影响。结果 与模型组比较，WJBF能显著延长大鼠扭体潜伏期，减少扭体次数（P<0.001）；显著降低卵巢及子宫指数（P<0.05、0.01、0.001）；显著降低大鼠血清PGF_2α、TXB2、E₂含量及PGF_2α/PGE₂，升高6-keto-PGF_1α、β-EP、Pg水平（P<0.05、0.01、0.001）；同时对子宫组织中PGF_2α、COX-2、Ca²⁺含量和PGF_2α/PGE₂有显著降低作用，对PGE₂水平有显著升高作用（P<0.05、0.01、0.001）。WJBF对离体子宫平滑肌自主收缩和激动剂诱导的强烈收缩子宫肌条的收缩频率、活动力、平均幅度具有显著抑制作用（P<0.05、0.01、0.001），且呈浓度相关性。结论 WJBF对原发性痛经模型大鼠具有明显改善作用，其机制可能与调节COX-2/PGF_2α通路及β-EP、E₂、Pg等内分泌激素的表达，抑制M受体的过度激活，进而缓解子宫平滑肌痉挛有关。

Objective To investigate the ameliorative effect and possible mechanism of action of Wuji Baifeng Soft Capsule（WJBF） on primary dysmenorrhea in rats. Methods SPF grade female SD rats were randomly divided into the control group, model group, ibuprofen sustained-release capsule(BLF, 0.08 g·kg^-1) group, Dingkun Dan(1.4 g·kg^-1) group and WJBF low, medium and high dose(0.3, 0.6, 1.2 g·kg^-1) groups, with 10 rats in each group. The rat model of primary dysmenorrhea was established by im of estradiol benzoate combined with intraperitoneal injection of oxytocin. From the 4 th day, each drug administration group was given the drug ig the corresponding dose for seven consecutive days. One hour after the last administration, the control group was intraperitoneally injected with 0.9% sodium chloride solution, and the other groups were intraperitoneally injected with oxytocin. The latency of writhing and the number of writhing within 30 min were recorded. The organ index of the uterus and ovaries was detected. The contents of prostaglandin F_2α(PGF_2α), prostaglandin E2（PGE₂）, thromboxane B₂（TXB₂）, 6-keto-prostaglandin F_1α(6-keto-PGF_1α), β-endorphin（β-EP）, 5-hydroxytryptamine（5-HT）, estradiol（E₂）, and progesterone（Pg） in rat serum were detected by enzyme-linked immunosorbent assay（ELISA）, and the ratio of PGF_2α/PGE₂ was calculated. The contents of PGF_2α, PGE₂, cyclooxygenase 2（COX-2）, and Ca²⁺ in uterine tissue were detected by ELISA or biochemical kits. Isolated uterine smooth muscle experiment: SPF-grade female rats were subcutaneously injected with 2.0 mg·kg^-1 estradiol benzoate for 2 consecutive days. On day 3, the rats were anesthetized, and uteri were rapidly excised to prepare 1 cm uterine rings. The specimens were suspended in an isolated organ perfusion device, aerated with 95% oxygen, and given a preload of 2.0 g. A multichannel physiological recorder was used to separately observe the effects of WJBF（0.37, 0.74, 1.48 mg·mL^-1） on spontaneous contraction, as well as acetylcholine（Ach）-and oxytocin-induced strong contractions of isolated uterine smooth muscle. Results Compared with the model group, WJBF significantly prolonged the latency of the convulsive response in rats, reduced the number of convulsions（P < 0.001）; significantly decreased the ovarian and uterine indices（P < 0.05, 0.01, 0.001）; significantly reduced the levels of serum PGF_2α, TXB₂, E₂ and PGF_2α/PGE₂, and elevated the levels of 6-keto-PGF_1α, β-EP, and Pg（P < 0.05, 0.01, 0.001）; at the same time, it significantly reduced the contents of PGF_2α, COX-2, Ca²⁺ in the uterine tissue and the ratio of PGF_2α/PGE₂, and significantly increased the level of PGE₂（P < 0.05, 0.01, 0.001）. WJBF had a significant inhibitory effect on the autonomous contraction of isolated uterine smooth muscle and the contraction frequency, activity, average amplitude, and activity force of the strongly contracted uterine strips induced by agonists（P < 0.05, 0.01, 0.001）, and showed a concentration-dependent relationship. Conclusion WJBF has a significant ameliorative effect on primary dysmenorrhea model rats, and the mechanism may be related to the regulation of COX-2/PGF_2α pathway and the expression of endocrine hormones, such as β-EP, E₂, Pg, etc., to inhibit the over-activation of M receptors, and then alleviate the spasm of uterine smooth muscle.

R965

国家科技重大专项-“四大慢病重大专项”(2025ZD0549700)