胶原蛋白肽联合海藻酸钠通过下调NLRP3炎症小体表达改善小鼠宫腔粘连
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1.新乡医学院;2.新乡医学院,新乡医学院三全学院;3.新乡医学院第三附属医院;4.南通业盟新材料股份有限公司

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【基金项目】国家重点研发计划资助(2018YFC1105800);南通业盟新材料股份有限公司业盟基金(110202002);新乡医学院第三附属医院开放课题(KFKTYB202127);新乡医学院研究生科研创新支持计划(YJSCX202124Z)。


Collagen peptide combined with sodium alginate improved intrauterine adhesions in mice by down-regulating the expression of NLRP3 inflammasome
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1.Xinxiang Medical University;2.Xinxiang Medical University,Sanquan College of Xinxiang Medical College;3.The Third Affiliated Hospital of Xinxiang Medical University;4.Nantong Yemeng New Material Co., Ltd

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Found by the National Key R & D Program Grant (2018YFC1105800), Nantong Yemeng New Material Co., Ltd Yemeng Foundation (110202002), Open Project of the Third Affiliated Hospital of Xinxiang Medical University (KFKTYB202127) and Postgraduate Scientific Research Innovation Support Program of Xinxiang Medical University (YJSCX202124Z).

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    摘要:

    【摘要】 目的 探讨胶原蛋白肽(collagen peptides,CP)联合海藻酸钠(sodium alginate, SA)对子宫内膜损伤的修复作用及机制。方法 将48只SPF级雌性C57BL/6N小鼠随机分为假手术组(sham)、模型组(model)、治疗组(CP SA),各16只。模型组采用微创法经小鼠子宫颈向宫腔注射95%乙醇造成子宫内膜损伤,制备小鼠宫腔粘连模型;假手术组向宫腔注射0.9%生理盐水;治疗组向宫腔注射95%乙醇后给予宫腔注射CP SA混合液。于造模7天后每组分别取8只小鼠采集样本用于各项检测。HE染色观察小鼠子宫组织病理形态学变化;改良Masson三色染色观察子宫内膜纤维化情况;Western blot检测子宫组织中炎症因子NLRP3、ASC、Caspase-1、IL-1β、IL-18蛋白表达水平;ELISA检测小鼠血清中炎症因子IL-1β、IL-18的表达。各组剩余的8小鼠按照雄雌比例为1:2合笼,记录各组小鼠妊娠数量,于妊娠第14天处死妊娠小鼠,记录胚胎数量。结果(1)与假手术组相比,模型组小鼠子宫内膜厚度变薄,子宫内膜纤维化增加,并有炎症细胞浸润。此外,模型组子宫组织中炎症因子NLRP3、ASC、Caspase-1、IL-1β和IL-18蛋白表达水平以及血清中炎症因子IL-1β和IL-18水平均显著高于假手术组(P<0.05)。(2)与模型组相比,治疗组小鼠子宫内膜损伤程度、子宫内膜纤维化程度及炎症细胞浸润均减轻;此外,治疗组子宫组织中炎症因子NLRP3、ASC、Caspase-1、IL-1β和IL-18蛋白表达水平及血清中炎症因子IL-1β和IL-18水平均显著低于模型组(P<0.05)。(3)与假手术组相比,模型组小鼠生殖能力下降(P<0.05);与模型组相比,治疗组小鼠生殖能力明显改善(P<0.05)。结论 CP和SA联合应用可有效改善宫腔粘连模型小鼠子宫组织病理改变,减轻子宫内膜损伤、炎症反应及纤维化程度,改善小鼠生殖能力,其作用机制可能与下调NLRP3炎症小体表达有关,为CP和SA联合应用治疗宫腔粘连提供可靠的实验依据。

    Abstract:

    【Abstract】Objective To investigate the repair effect and mechanism of collagen peptides (CP) combined with sodium alginate (SA) on endometrial injury. Methods Forty-eight SPF female C57BL/6N mice were randomly divided into sham group (16 mice), model group (16 mice) and CP SA group (16 mice). In the model group, 95% ethanol was injected into the uterine cavity through the cervix to induce endometrial injury, and the intrauterine adhesion model was established. The sham operation group was injected with 0.9% normal saline into the uterine cavity. The treatment group was injected with the mixture of CP SA after the injection of 95% ethanol into the uterine cavity. After 7 days of modeling, 8 mice in each group were selected to collect samples for each test. HE staining was used to observe the pathological changes of mouse uterine tissue. Modified Masson tri-color staining was used to observe endometrial fibrosis. Western blot was used to detect the protein expression levels of inflammatory factors NLRP3, ASC, Caspase-1, IL-1β and IL-18 in uterine tissues. ELISA was used to detect the expression of IL-1β and IL-18 in serum. The remaining 8 mice in each group were cohoused according to the male-female ratio of 1:2, and the number of pregnant mice in each group was recorded. Pregnant mice were euthanized in the 14th day of pregnancy, and the number of embryos was recorded. Results (1) Compared with the sham operation group, the model group showed thinner endometrial thickness, increased endometrial fibrosis, and inflammatory cell infiltration. In addition, the protein expression levels of inflammatory factors NLRP3, ASC, Caspase-1, IL-1β and IL-18 in uterine tissue and the levels of IL-1β and IL-18 in serum in the model group were significantly lower than those in the sham operation group (P<0.05). (2) Compared with the model group, the treatment group showed that the degree of endometrial injury, endometrial fibrosis and inflammatory cell infiltration were reduced. In addition, the protein expression levels of inflammatory factors NLRP3, ASC, Caspase-1, IL-1β and IL-18 in uterine tissue and the levels of IL-1β and IL-18 in serum in the model group were significantly decreased than those in the sham operation group (P< 0.05). (3) Compared with the sham group, the reproductive ability of the model group decreased (P<0.05). Compared with the model group, the reproductive ability of the treatment group was significantly improved(P<0.05). Conclusions The combined application of CP and SA can effectively improve the pathological changes of uterine tissue, reduce endometrial injury, inflammatory response and fibrosis, and improve the reproductive ability of mice. The mechanism may be related to the down-regulation of NLRP3 inflammasome expression, which provides a reliable experimental basis for the combined application of CP and SA in the treatment of intrauterine adhesion.

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  • 收稿日期:2023-05-03
  • 最后修改日期:2023-09-15
  • 录用日期:2023-09-15
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