| 乌兰,李心竹,黄露诗.养血化瘀合剂调控血管通透性防治卵巢过度刺激综合征作用及机制研究[J].江苏中医药,,():. |
| 养血化瘀合剂调控血管通透性防治卵巢过度刺激综合征作用及机制研究 |
| Stucia-removing herbs prevent and treat Ovarian Hyperstimulation Syndrome by Regulating vascular permeability |
| 投稿时间：2023-12-25 修订日期：2024-04-22 |
| DOI： |
| 中文关键词: 养血化瘀合剂 卵巢过度刺激综合征 血管通透性 VEGF PEDF NLRP3 |
| 英文关键词: Stucia-removing herbs ovarian stress syndrome (OHSS) vascular permeability VEGF PEDF NLRP3 |
| 基金项目:其它, |
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| 中文摘要: |
| 目的：探讨养血化瘀合剂对卵巢应激综合征（OHSS）小鼠的预防治疗作用及其机制。方法：将72只雌性ICR小鼠随机分为模型组、养血化瘀合剂低、中、高剂量组（分别每日灌胃给药1.42、2.84、5.68 g/kg）、吲哚美辛组（每日灌胃给药的剂量为7.5 mg/kg）、正常对照组（每日仅给予等体积生理盐水灌胃），每组12只，连续给药7天。从给药第2天开始采用人绒毛膜促性腺激素（Human Chorionic Gonadotropin, HCG）和孕马血清促性腺激素（Pregnant Mare Serum Gonadotropin, PMSG）联用的方法构建OHSS模型，正常对照组只注射等体积生理盐水，第7天处死全部小鼠，处死前30分钟每组随机选取3只小鼠予尾静脉注射伊文思蓝，并进行血管通透性检测。对所有小鼠进行体重称量，麻醉，眼球取血，血清一氧化氮（NO）含量测定，采用酶联免疫吸附测定（ELISA）法测定白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）、血管内皮生长因子（Vascular Endothelial Growth Factor, VEGF）、孕激素（progestogen, P）以及雌二醇（Estradiol, E2）的水平。处死全部小鼠，取完整卵巢称重，采用苏木精-伊红染色（HE）法观察各组小鼠卵巢组织的病理变化；采用蛋白免疫印迹（Western Blot, WB）法检测VEGF，色素上皮衍生因子（Pigment Epithelium Derived Factor, PEDF），半胱氨酸天冬氨酸酶（caspase-1）及、NOD样受体热蛋白结构域相关蛋白3（NLRP3）的蛋白表达量。结果：OHSS模型组小鼠体重、卵巢体积、重量相比于正常对照组明显增加（P<0.01），卵巢组织内黄体的数量增加，同时形成多个卵泡；养血化瘀合剂低、中剂量组和吲哚美辛组体重明显高于模型组（P<0.01），其中养血化瘀合剂低剂量组的卵巢重量显著低于模型组（P<0.05），养血化瘀合剂和吲哚美辛组的卵巢组织体积均显著缩小，同时黄体和卵泡数量相比模型组明显减少。OHSS模型组小鼠血清中IL-6、THF-α、VEGF、P和E2水平明显高于正常对照组（P<0.01），血清中NO水平也明显高于正常对照组（P<0.01），卵巢浸出液中的伊文思蓝含量显著增加（P<0.05）；养血化瘀合剂高剂量组和吲哚美辛组小鼠血清中IL-6、THF-α、VEGF水平明显低于模型组（P<0.01），中剂量组小鼠血清中NO、VEGF、THF-α、IL-6水平也明显低于模型组（P<0.01，P<0.05，P<0.01），低剂量组小鼠血清的NO、E2和P水平明显低于模型组（P<0.01，P<0.01）。OHSS模型组小鼠卵巢组织中VEGF，Caspase-1及NLRP3的蛋白表达量相比于对照组明显增加，PEDF的表达量则明显低于对照组；在养血化瘀合剂低、中剂量组中VEGF，Caspase-1及NLRP3的蛋白表达量呈上升趋势，PEDF的表达量则呈下降趋势。结论：养血化瘀合剂对OHSS起到一定防治效果，中剂量较优，能够有效降低OHSS小鼠血管通透性，且对小鼠体重、卵巢重量、腹水有明显的改善作用，可能通过调节NLRP3/caspase-1信号通路，调控OHSS小鼠血清中NO、炎症因子和激素的水平，起到防治该疾病的效果。 |
| 英文摘要: |
| Objective To investigate the therapeutic effect and mechanism of different doses of stay-removing Chinese medicine on ovarian stress syndrome (OHSS) model mice. Methods Sixty 5-week-old female ICR mice were randomly divided into 6 different groups: blank control group, OHSS model group, low, medium and high doses of Chinese medicine groups, and positive drug control group (indomethacin), with 10 mice in each group. Except for the blank control group, the other 5 groups were imposed with the combination of pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin (HCG) to construct OHSS model. At the same time, normal saline, different doses of Yangxue Huayu recipe and indomethacin were given respectively. After 8 days of continuous administration, the mice were injected with Evans Blue (EB) through the tail vein for vascular permeability detection. Then body weight and ovarian weight were measured, and the levels of interleukin-6 (IL-6), estradiol (E2), progesterone (P) tumor necrosis factor-α (TNF-α) and vascular endothelial growth factor (VEGF) were determined by ELISA. The expression of nucleotide-binding oligomerization domain, leucine-rich repeat and pyrin domain-containing 3 (NLRP3), cysteinyl aspartate specific proteinase (caspase-1), VEGF and pigment epithelium-derived factor (PEDF) are detected with Western Blot (WB). Results The ovarian weight, body weight, levels of NO,VEGF, IL-6 and THF-α, E2 and P in serum are all dramatically decreased (P<0.05) after the administration of Yangxue Huayu combination, which cause the reduction of vascular permeability in OHSS mice. The expression of PEDF increased following the application of low and medium doses of Yangxue Huayu combination, according to WB data, however the expressions of VEGF, caspase-1, and NLRP3 considerably decreased. Conclusion Yangxue Huayu combination can effectively regulate the level of some inflammatory factors in serum of OHSS mice, reduce vascular permeability, relieve ascites, weight of body and ovarian, and play an effective role in preventing and treating the disease. The possible mechanism of action is through the regulation of VEGF, PEDF, caspase-1 and NLRP3 pathways. |
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