Xue Xiao, Liu Yu, Wang Shao-Hua, Yuan Han-Yu, Li Juan, Pan Si-An, Yue Zeng-Hui
School of Acupuncture-moxibustion and Massage, Hunan University of Chinese Medicine, Changsha 410208, China; Department of Traditional Chinese Medicine, The First Affiliated Hospital of University of South China, Hengyang 421001, Hunan Province.
School of Acupuncture-moxibustion and Massage, Hunan University of Chinese Medicine, Changsha 410208, China.
Zhen Ci Yan Jiu. 2023 Jan 25;48(1):63-70. doi: 10.13702/j.1000-0607.20220224.
To investigate the mechanism of electroacupuncture(EA) intervention in rats with primary dysmenorrhea(PDM) based on the Toll-like receptor 4(TLR4)/nuclear factor(NF)-κB signaling pathway.
Forty female SD rats were randomly divided into blank control, model, EA and medication groups, with 10 rats in each group. PDM rat model was established by subcutaneous injection of estradiol benzoate combined with intraperitoneal injection of oxytocin. At the same time of model procedures, EA(50 Hz, dense wave) was applied to "Guanyuan" (CV4) and bilateral "Sanyinjiao" (SP6) of rats in the EA group, with needles retained for 20 min, for 10 consecutive days. Rats in the medication group received ibuprofen(125 mg/100 mL, 0.8 mL) by gavage for 10 consecutive days. At the 11th day, writhing behavior of rats was assessed. Uterine morphology was observed by eyes and uterine pathological changes were observed after HE staining. Content of prostaglandin E2 (PGE2) and prostaglandin F2α (PGF2α) in serum and uterine tissues was detected by ELISA; NF-κB p65 positive expression in nucleus was detected by immunofluorescence; protein expression levels of TLR4, NF-κB p65, p-NF-κB p65 and inflammatory factors interleukin (IL) -1β and IL-18 were detected by Western blot.
After modeling, uterus tissues were congested and edematous, with necrosis of luminal epithelium, severe edema and extensive shedding of endometrium, nuclear pyknosis, fragmentation and disappearance, neutrophils infiltration, and slight expansion of glandular cavity, which was milder in the EA and the medication groups. Compared with the blank control group, writhing times, scores and incubation period, HE pathological scores, PGF2α contents in serum and uterine tissues, ratio of NF-κB p65 positive expression in nucleus, TLR4, NF-κB p65, p-NF-κB p65, IL-1β and IL-18 protein expression levels in uterine tissues of rats in the model group were all significantly increased(<0.01), while PGE2 contents in serum and uterine tissues were significantly decreased(<0.01). Compared with the model group, writhing times and scores, HE pathological scores, PGF2α contents in serum and uterine tissues, ratio of NF-κB p65 positive expression in nucleus, TLR4, NF-κB p65, p-NF-κB p65, IL-1β and IL-18 protein expression levels in uterine tissues of rats in the EA and medication group were all significantly decreased(<0.01), while writhing incubation period, PGE2 contents in serum and uterine tissues were significantly increased(<0.05, <0.01).
EA intervention could relieve inflammatory response and pain in PDM rats, which may be related to its effect in reducing TLR4 expression, inhibiting NF-κB activation and down-regulating inflammatory factors levels of IL-1β and IL-18.
基于Toll样受体4（TLR4）/核因子（NF）-κB信号通路探讨电针干预原发性痛经（PDM）大鼠的机制。
将40只雌性SD大鼠随机分为空白对照组、模型组、电针组和药物组，每组10只。采用皮下注射苯甲酸雌二醇联合腹腔注射缩宫素的方法建立PDM大鼠模型。在造模的同时，对电针组大鼠的“关元”（CV4）和双侧“三阴交”（SP6）施加电针（50Hz，密波），留针20min，连续10天。药物组大鼠连续10天灌胃给予布洛芬（125mg/100mL，0.8mL）。第11天，评估大鼠的扭体行为。肉眼观察子宫形态，HE染色后观察子宫病理变化。采用ELISA法检测血清和子宫组织中前列腺素E2（PGE2）和前列腺素F2α（PGF2α）的含量；采用免疫荧光法检测细胞核中NF-κB p65的阳性表达；采用蛋白质印迹法检测TLR4、NF-κB p65、p-NF-κB p65蛋白表达水平及炎症因子白细胞介素（IL）-1β和IL-18的表达水平。
造模后，子宫组织充血水肿，管腔上皮坏死，子宫内膜严重水肿、广泛脱落，细胞核固缩、碎裂、消失，中性粒细胞浸润，腺腔轻度扩张，电针组和药物组病变较轻。与空白对照组比较，模型组大鼠的扭体次数、评分及潜伏期、HE病理评分、血清和子宫组织中PGF2α含量、细胞核中NF-κB p65阳性表达率、子宫组织中TLR4、NF-κB p65、p-NF-κB p65、IL-1β和IL-18蛋白表达水平均显著升高（<0.01），而血清和子宫组织中PGE2含量显著降低（<0.01）。与模型组比较，电针组和药物组大鼠的扭体次数、评分、HE病理评分、血清和子宫组织中PGF2α含量、细胞核中NF-κB p65阳性表达率、子宫组织中TLR4、NF-κB p65、p-NF-κB p65、IL-1β和IL-18蛋白表达水平均显著降低（<0.01），而扭体潜伏期、血清和子宫组织中PGE2含量显著升高（<0.05，<0.01）。
电针干预可减轻PDM大鼠的炎症反应和疼痛，其机制可能与降低TLR4表达、抑制NF-κB活化及下调IL-1β和IL-18炎症因子水平有关。
