富血小板血浆对驴感染性子宫内膜炎的体外抑菌效果研究

doi:10.16431/j.cnki.1671-7236.2025.07.047

摘要/Abstract

摘要： 【目的】探究富血小板血浆(platelet rich plasma，PRP)对驴感染性子宫内膜炎的抗炎和抑菌效果。【方法】选用健康驴的子宫内膜作为外植体进行体外培养，添加1.82×10⁷ CFU/mL马兽疫链球菌亚种(Streptococcus equi subsp. zooepidemicus，S.zooepidimicus)，诱导驴感染性子宫内膜炎外植体模型。试验分为7组，分别为空白对照组(MCG)、模型组(MG)、阳性药物对照组(PDCG)、25% PRP组、50% PRP组、MIX1组和MIX2组。试验结束后收集各组培养液和外植体，对培养液进行细菌培养和白细胞介素-1(IL-1)、IL-6浓度检测，对外植体进行组织学检查，比较各组培养液中细菌浓度、外植体组织学炎性变化和炎性因子水平，探究PRP的抑菌和抗炎效果。【结果】试验制备所得的PRP血小板浓度为4.69×10¹¹/L，血小板生长因子(PDGF)浓度为32.01 ng/mL，转化生长因子-β1(TGF-β1)浓度为49.37 ng/mL。模型组培养液中细菌浓度极显著高于空白对照组(P＜0.01)，25% PRP、50% PRP、MIX1和MIX2组培养液细菌浓度极显著低于模型组(P＜0.01)。空白对照组外植体子宫内膜结构完整，多形核白细胞少量分布；模型组子宫内膜结构严重破坏，多形核白细胞大量浸润；与模型组相比，PRP处理组子宫内膜腔上皮细胞和腺体结构更完整，多形核白细胞浸润面积较小。模型组培养液中IL-1和IL-6浓度极显著高于空白对照组(P＜0.01)，PRP处理后培养液中IL-1和IL-6浓度极显著低于模型和阳性药物对照组(P＜0.01)。【结论】本研究成功构建了马兽疫链球菌亚种诱导的驴感染性子宫内膜炎外植体模型，PRP可显著抑制外植体模型中细菌生长，下调炎性因子IL-1和IL-6的表达，减缓外植体组织炎性病变。

关键词: 富血浆血小板(PRP); 驴; 感染性子宫内膜炎; 外植体

Abstract: 【Objective】 This study aimed to investigate the anti-inflammatory and antibacterial effects of platelet rich plasma (PRP) on infectious endometritis in donkeys. 【Method】 Healthy donkey endometrium was selected as explants for in vitro culture,and 1.82×10⁷ CFU/mL Streptococcus equi subsp. zooepidemicus (S. zooepidimicus) was added to induce an infection-based endometritis explant model.The experiment was divided into 7 groups,which were mock control group (MCG),model group (MG),positive drug control group (PDCG),25% PRP and 50% PRP groups,MIX1 and MIX2 group.After the experiment,the culture medium and explants of each group were collected,and the concentration of interleukin-1 (IL-1) and IL-6 were detected in the culture medium,and the explants were examined by histology.The antibacterial and anti-inflammatory effects of PRP were evaluated by comparing bacterial concentrations in culture media,histopathological inflammatory changes in explants,and the levels of inflammatory factors. 【Result】 The PRP platelet concentration was 4.69×10¹¹/L,the platelet-derived growth factor (PDGF) concentration was 32.01 ng/mL,and the transforming growth factor β1 (TGF-β1) concentration was 49.37 ng/mL.The bacterial concentration in culture medium of MG group was extremely significantly higher than that of MCG group (P<0.01),and the bacterial concentrations in culture medium of 25% PRP,50% PRP,MIX1 and MIX2 groups were extremely significantly lower than that of MG group (P<0.01).In MCG group,the endometrium structure of the explants was intact,and few polymorphonuclear leukocytes were distributed.In MG group,the endometrial structure was severely damaged and polymorphonuclear leukocytes were infiltrated.Compared with MG group,PRP treated groups had more complete epithelial and glandular structures and less infiltration area of polymorphonuclear leukocytes.The concentration of IL-1 and IL-6 in culture medium of MG group was extremely significantly higher than that of MCG group (P<0.01).The concentrations of IL-1 and IL-6 in culture medium treated with PRP were extremely significantly lower than those in MG and PDCG groups (P<0.01). 【Conclusion】 In this study,an explants model of donkey infective endometritis induced by S. zooepidimicus was successfully constructed.PRP could significantly inhibit the growth of bacteria in explants,down-regulate the expression of inflammatory factors IL-1 and IL-6,and alleviate tissue inflammation in the explants.

Key words: platelet rich plasma (PRP); donkey; infectious endometritis; explant

中图分类号:

S852.2

兰芯儿, 刘思睿, 陈庆琳, 蒋辉, 彭聪, 王子璇, 朱怡平, 李靖. 富血小板血浆对驴感染性子宫内膜炎的体外抑菌效果研究[J]. 中国畜牧兽医, 2025, 52(7): 3489-3495.

LAN Xiner, LIU Sirui, CHEN Qinglin, JIANG Hui, PENG Cong, WANG Zixuan, ZHU Yiping, LI Jing. Inhibitory Effect of Platelet Rich Plasma on Infectious Endometritis in Donkeys in vitro[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(7): 3489-3495.

参考文献

[1] TROEDSSON M H T,WOODWARD E M.Our current understanding of the pathophysiology of equine endometritis with an emphasis on breeding-induced endometritis[J].Reproductive Biology,2016,16(1):8-12.
[2] BENKO T,BOLDIZAR M,NOVOTNY F,et al.Incidence of bacterial pathogens in equine uterine swabs,their antibiotic resistance patterns,and selected reproductive indices in English thoroughbred mares during the foal heat cycle[J].Veterinární Medicína,2015,60(11):613-620.
[3] FERRER M S,PALOMARES R.Aerobic uterine isolates and antimicrobial susceptibility in mares with post-partum metritis[J].Equine Veterinary Journal,2018,50(2):202-207.
[4] ZHAO Y,ZHU Y,LIU B,et al.Antimicrobial susceptibility of bacterial isolates from donkey uterine infections,2018-2021[J].Veterinary Sciences,2022,9(2):67.
[5] SEGABINAZZI L G T M,CANISSO I F,PODICO G,et al.Intrauterine blood plasma platelet-therapy mitigates persistent breeding-induced endometritis,reduces uterine infections,and improves embryo recovery in mares[J].Antibiotics (Basel,Switzerland),2021,10(5):490.
[6] LIU X,WANG Y,WEN X,et al.Platelet rich plasma alleviates endometritis induced by lipopolysaccharide in mice via inhibiting TLR4/NF-κB signaling pathway[J].American Journal of Reproductive Immunology,2024,91(3):e13833.
[7] DE BARROS M R M,DAVIES-MOREL M C G,MUR L A J,et al.Characterization of an ex vivo equine endometrial tissue culture model using next-generation RNA-sequencing technology[J].Animals,2021,11(7):1995.
[8] HARRISON T E,BOWLER J,LEVINS T N,et al.Platelet yield and yield consistency for six single-spin methods of platelet rich plasma preparation[J].Platelets,2020,31(5):661-666.
[9] SHIN H S,WOO H M,KANG B J.Optimisation of a double-centrifugation method for preparation of canine platelet-rich plasma[J].BMC Veterinary Research,2017,13(1):198.
[10] HAUSCHILD G,GEBUREK F,GOSHEGER G,et al.Short term storage stability at room temperature of two different platelet-rich plasma preparations from equine donors and potential impact on growth factor concentrations[J].BMC Veterinary Research,2017,13(1):7.
[11] CARMONA J U,RÍOS D L,LÓPEZ C,et al.Proinflammatory and anabolic gene expression effects of platelet-rich gel supernatants on equine synovial membrane explants challenged with lipopolysaccharide[J].Veterinary Medicine International,2017,2017:6059485.
[12] BORGES A M,HEALEY G D,SHELDON I M.Explants of intact endometrium to model bovine innate immunity and inflammation ex vivo[J].American Journal of Reproductive Immunology,2012,67(6):526-539.
[13] WEI S,XU P,YAO Z,et al.A composite hydrogel with co-delivery of antimicrobial peptides and platelet-rich plasma to enhance healing of infected wounds in diabetes[J].Acta Biomaterialia,2021,124:205-218.
[14] WESSELY-SZPONDER J,SZPONDER T,BOBOWIEC R,et al.Changes in the activity of ovine blood-derived macrophages stimulated with antimicrobial peptide extract (AMP) or platelet-rich plasma (PRP)[J].Journal of Veterinary Research,2019,63(2):235-242.
[15] GHALLAB R S,EL-BESKAWY M,EL-SHEREIF A A,et al.Impact of intrauterine infusion of platelets-rich plasma on endometritis and reproductive performance of Arabian mare[J].Reproduction in Domestic Animals,2023,58(5):622-629.
[16] MUIR A J T,NIEHAUS A J,LOZIER J W,et al.Autologous platelet-rich plasma effects on Staphylococcus aureus-induced chondrocyte death in an in vitro bovine septic arthritis model[J].American Journal of Veterinary Research,2021,83(2):119-126.
[17] WU R,YU F,HOLYOAK G R,et al.Use of the endometrial histopathology to improve diagnosis of donkeys with endometritis[J].Equine Veterinary Education,2024,36(1):43-50.
[18] LI C,WANG J,YANG W,et al.3D-printed hydrogel particles containing PRP laden with TDSCs promote tendon repair in a rat model of tendinopathy[J].Journal of Nanobiotechnology,2023,21(1):177.
[19] TIAN K,YE J,ZHONG Y,et al.Autologous i-PRF promotes healing of radiation-induced skin injury[J].Wound Repair and Regeneration,2023,31(4):454-463.
[20] BENDINELLI P,MATTEUCCI E,DOGLIOTTI G,et al.Molecular basis of anti-inflammatory action of platelet-rich plasma on human chondrocytes:Mechanisms of NF-κB inhibition via HGF[J].Journal of Cellular Physiology,2010,225(3):757-766.
[21] CHEN S,ZHU L,FANG X,et al.Alloferon mitigates LPS-induced endometritis by attenuating the NLRP3/CASP1/IL-1β/IL-18 signaling cascade[J].Inflammation,2024.Doi:10.1007/s10753-024-02083-6.Online ahead of print.
[22] MISHRA S,SAHU S K,PANIGRAHI S,et al.Comparative therapeutic efficacy between levofloxacin,ornidazole and alpha tocopherol combination to prostaglandin F2α on IL-6 and IL-10 transcript level in longstanding cases of endometritis in crossbreed Jersey cows[J].Iranian Journal of Veterinary Research,2018,19(3):217-224.
[23] ELSAYED D H,EL-AZZAZI F E,MAHMOUD Y K,et al.Subclinical endometritis and postpartum ovarian resumption in respect to TNF-α,IL-8 and CRP in Egyptian buffaloes[J].Animal Reproduction,2020,17(1):e20190027.