METRNL在PCOS患者血清中的表达及意义.pdf

.1342生殖医学杂志2 0 2 3年9 月第32 卷第9 期DO1:10.3969/j.issn.1004-3845.2023.09.007METRNL在PCOS 患者血清中的表达及意义任小胖，郝婉姣，刘梅云，宋洁，李烨，薛会灵*（河北大学附属医院生殖医学科，保定0 7 10 0 0）【摘要】目的探讨多囊卵巢综合征(PCOS)患者血清中METRNL的表达情况，寻找PCOS治疗的新靶点。方法选取2022年7-10 月就诊于河北大学附属医院生殖中心门诊被确诊为PCOS的131例患者为实验组（PCOS组），另收集同期确诊为男方因素不孕的且其他各项实验室检查结果均正常的50 例女性作为正常对照组。应用酶联免疫吸附(ELISA)法检测血清中METRNL水平，化学发光法检测血清中睾酮（T)、卵泡刺激素（FSH)、黄体生成素（LH)水平；对所有研究对象进行胰岛素释放实验（IRT)及葡萄糖耐量实验（OGTT)。同时根据体质量指数（BMI)将PCOS组分为肥胖组及非肥胖组；根据胰岛素抵抗指数（HOMA-IR)将PCOS组分为胰岛抵抗组（PCOS-IR组）与非胰岛抵抗组（PCOS-非IR组），比较PCOS及PCOS亚组与对照组之间的METRNL、激素水平等差异，并用Pearson分析各指标间的相关性。结果PCOS组与对照组比较,BMI、T、LH/FSH、H O M A-I R 及METRNL水平均有统计学差异（P0.05）。PC O S肥胖组患者的HOMA-IR水平显著高于非肥胖者（P0.05），M ET R NL的表达水平显著低于非肥胖PCOS患者（P0.05）；与对照组比较,PCOS非肥胖组的T、LH/FSH及HOMA-IR值均显著升高（P0.05），而METRNL的表达水平显著降低（P0.05）。PC O S-I R 组的BMI水平显著高于PCOS-非IR组（P0.05）,M ET R NL的表达水平显著低于PCOS-非IR组（P0.05）；与对照组比较,PCOS-非IR组的T、LH/FSH均显著升高（P0.05），M ET R NL的表达水平显著降低（P0.05）。Pe a r s o n 相关分析结果显示，所有研究对象中METRNL与HOMA-IR、BM I、T 及LH/FSH均呈负相关（P0.05），而在PCOS组中，METRNL与HOMA-IR、BM I 均呈负相关(P0.05),与T及LH/FSH无相关性。结论血清中METRNL水平的变化及其与HOMA-IR、BM I、T 及LH/FSH的关系，可能与PCOS的发生有关，有望成为PCOS治疗的新靶点。【关键词】多囊卵巢综合征；METRNL；体质量指数；胰岛素抵抗【中图分类号】R711.6Expression and significance of METRNL in serum of PCOS patientsREN Xiao-pang,HAO Wan-jiao,LIUMei-yun,SONG Jie,LI Ye,XUE Hui-lingDepartment of Reproductive Medicine,Af filiated Hospital of Hebei University,Baoding 071000Objective:To investigate the serum expression of METRNL in the patients with polycystic ovariansyndrome(PCOS),and to find a new therapeutic target for PCOS.Methods:A total of 131 patients who were diagnosed as PCOS at the Clinic of Reproductive Center ofAffiliated Hospital of Hebei University from July to October 2022 were selected as the experimental group(PCOS group),and another 50 women who were diagnosed with male infertility during the same period andhad normal laboratory test results were collected as the normal control group.The enzyme-linkedimmunosorbent assay(ELISA)was used to detect serum METRNL levels.The serum levels of FSH,LHand testosterone(T)were detected by chemiluminescence method.Oral glucose tolerance test(OGTT)andinsulin release test(IRT)were performed for all enrolled subjects.The patients in PCOS group weredivided into an obese group and a non-obese group according to body mass index(BMI),and the patients【文献标识码】A【A b s t r a c t【收稿日期】2023-02-08；【修回日期】2 0 2 3-0 4-0 9【基金项目】河北大学附属医院青年基金项目（2 0 2 1Q044)【作者简介】住任小胖，女，河北保定人，硕士，生殖医学专业，（*通讯作者）生殖医学杂志2 0 2 3年9 月第32 卷第9 期were divided into the insulin resistance(IR)group and the non-IR group according to HOMA-IR index.The differences in METRNL and hormone levels between PCOS,PCOS subgroups and the control groupwere compared,and Pearson was to analyze the correlation between each indicator.Results:There were significant differences in BMI,T,LH/FSH,HOMA-IR,and METRNL levelsbetween the PCOS group and the control group(P0.05).The HOMA-IR levels in PCOS obese patientswere significantly higher than those in non-obese patients(P0.05).Compared with the control group,the levels of T&LH/FSH,and HOMA-IR values of the non-obese PCOS group were significantlyincreased(P0.05),while the METRNL expression level was significantly reduced(P0.05).The BMIlevel of the PCOS-IR group was significantly higher than that of the PCOS-non-IR group(P.O5),whilethe METRNL expression level was significantly lower than that of the PCOS-non-IR group(P0.05).Compared with the control group,the levels of T,LH/FSH in the PCOS non-IR group were significantlyincreased(P0.05),while the METRNL expression level was significantly reduced(P0.05).Pearsoncorrelation analysis showed that METRNL was negatively correlated with HOMA-IR,BMI,T and LH/FSH in all patients(P0.05),while METRNL was negatively correlated with HOMA-IR and BMI(P 10 ml)；(2)临床或生化检测的睾酮（T)升高（本中心实验指标：T0.48 nmol/L)；（3)无排卵性月经失调；符合上.1344：述3项中2 项,并排除其它病因者可以诊断为PCOS。正常对照组：（1)月经规则，无肥胖、多毛和/或痤疮等表现；（2)生殖激素及内分泌实验室测定正常；(3)B超检测提示卵巢无多囊改变、无子宫肌瘤及子宫内膜异位症声像改变；（4)无发热及炎症性疾病。所有入组患者均排除高泌乳素血症、全身炎症性疾病、甲状腺、肾上腺疾病、高血压、糖尿病等合并症及并发症，并至少3个月内未用过任何激素类药物。本研究获得河北大学附属医院伦理委员会的批准，所有研究对象均签署知情同意书。二、研究方法1.分组:PCOS组患者按照BMI值分为非肥胖组与肥胖组；根据HOMA-IR将PCOS组进行分为PCOS-IR组和 PCOS-非 IR组。参数计算：（1)胰岛素抵抗指数（HOMA-IR）：根据空腹胰岛素（FINS)和空腹血糖(FPG)值,计算自然人群中的HOMA-IR，即HOMA-IR=（FPG X FI NS)/2 2.5，以人群分布的上1/4位点作出估测IR的切点，其数值为2.6 910-11，HOMA-IR2.69为IR。（2)体质量指数(BMI)：BMI28 kg/m为肥胖。2.血清生殖激素测定：所有人组对象均于月经周期第2 4日或闭经2 个月以上、且经阴超检查无优势卵泡时，空腹采集静脉血5ml,应用化学发光法检测睾酮（T）、卵泡刺激素（FSH)、黄体生成素（LH）、雌二醇（E,）、孕酮（P）、泌乳素（PRL)水平。3.血清METRNL的测定：采集空腹静脉血5ml，2500rpm离心2 0 min后，将标本置于一2 0 冻存备用。应用ELISA法测定血清METRNL水平，试组别例数对照组50PCOS 组131注：与对照组比较，*P0.05）;PC O S肥胖组患者的HOMA-IR水平显著高于非肥胖者（P0.05），生殖医学杂志2 0 2 3年9 月第32 卷第9 期剂盒购自上海瑞番生物有限公司，具体操作方法严格按照产品说明书进行。4.葡萄糖耐量实验（OGTT)及胰岛素释放实验（IRT）：所有对象均于早8 时空腹行OGTT及IRT测定,采集空腹静脉血后口服葡萄糖7 5g，分别于服糖后30 min、6 0 m in、12 0 m in 采集静脉血，应用氧化酶法测定血清葡萄糖(SG)水平,应用化学发光法测定血清胰岛素（INS)水平，以获得FPG及FINS数据，计算 HOMA-IR进行分析,并排除糖尿病患者。5.研究指标:所有人组患者血清T、FSH、LH、E、P、PR L、M ET R NL、FI NS 和 FPG 水平;LH/FSH以及 HOMA-IR。三、统计学分析采用SPSS16.0统计软件进行数据分析。正态分布的计量数据采用均数土标准差（士s)表示，组间比较采用t检验；非正态分布数据进行对数转换以符合正态分布，再进行分析；采用Pearson进行相关性分析。P 0.0 5），其余指标如 BMI、T、LH/FSH、HOMA-IR、M ET R NL相比均有统计学差异（P0.05)(表 1)。表1PCOS组与对照组各项指标比较(士s)年龄BMI/(kg/m)27.484.1522.103.9027.354.8626.565.04*结果T/(nmol/L)LH/FSH0.260.120.710.410.460.17*1.770.32*METRNL的表达水平显著低于非肥胖PCOS患者(P0.05)；与对照组比较,PCOS非肥胖组的T、LH/FSH及HOMA-IR值均显著升高（P0.05)，而METRNL的表达水平显著降低（P0.05）(表2)。HOMA-IRMETRNL/(pg/ml)2.130.9910.971.734.312.62*4.621.74*(pg/ml)生殖医学杂志2 0 2 3年9 月第32 卷第9 期组别年龄对照组5027.484.15PCOS 组非肥胖组82肥胖组4927.324.68注：与非肥胖组比较，*P0.05）；PCO S-IR 组的BMI水平显著高于PCOS-非表3PCOS患者根据HOMA-IR分组后各项指标与对照组的比较(元士s)组别例数对照组5027.484.15PCOS 组 PCOS-非 IR30PCOS-IR101注：与PCOS-非IR组比较，*P0.05。四、METRNL与各项指标的相关性Pearson相关性分析结果显示，所有研究对象中METRNL与BMI(r=-0.294,P=0.000)、LH/FSH值（r=0.066,P=0.000）、I R（r=0.284,P=0.000)、T(r=0.318,P=0.0 0 0)均呈负相关，与年龄无明显相关（r=一0.0 7 2,P=BMIPMETRNL一0.2 9 40.000*注：*P0.05。表5PCOS组中METRNL与各项指标的相关性BMIHOMA-IRPPMETRNL0.1750.046*注：P0.05。1345表2 PCOS患者根据BMI分组各项指标与对照组的比较(士s)METRNL/例数BMI/(kg/m)22.103.9027.364.9223.312.7131.922.79*年龄BMI/(kg/m)22.13.9027.135.4522.773.1127.414.6427.654.91*表4METRNL与各项指标的相关性分析HOMA-IRP-0.2840.0000.1910.029*T/(nmol/L)0.260.12*0.470.180.440.16IR组（P0.05）,M ET R NL的表达水平显著低于PCOS-非IR组（P0.05）；与对照组比较，PCOS-非IR组的T、LH/FSH 均显著升高（P0.05）M ET R NL的表达水平显著降低（P0.05)(表 3)。METRNL/T/(nmol/L)LH/FSH0.260.12*0.710.41*0.470.181.960.870.450.171.87 1.130.365)(表 4)。在 PCOS 组中,METRNL 与 BMI(r=一0.17 5,P=0.046)、I R（r=0.19 1,P=0.0 2 9)均呈负相关，与年龄（r=一0.136，P=0.123）、LH/FSH 值(r=-0.132,P=0.131)、T(r=0.0 6 4,P=0.470）无明显相关（表5）。LH/FSHTPP-0.0660.000*LH/FSHP-0.1320.131LH/FSH0.710.41*1.730.271.810.34-0.3180.000*TP一0.0 6 40.470HOMA-IR2.130.99*10.971.73*3.512.224.861.645.582.73*4.271.30*HOMA-IR(pg/ml)2.130.9910.971.73*1.880.565.621.694.992.56*4.341.38*年龄P-0.0720.365年龄P-0.1360.123.1346PCOS是生育期妇女常见的一种复杂的内分泌及代谢异常所致疾病,其发病率日趋增加,但其发病机制目前尚不明确。研究表明，多种炎症因子在PCOS中表达异常，表明 PCOS可能是一种低度慢性炎症,此外PCOS患者机体组织对胰岛素敏感性下降，通常表现为对糖代谢的效应下降，葡萄糖不能得到充分利用，导致血糖升高，胰岛素分泌代偿性增加,进而导致高胰岛素血症。由此推断慢性炎症、IR和伴随的高胰岛素血症在PCOS的发病机制中起到关键的作用。METRNL由活化的巨噬细胞或由巨噬细胞集落刺激因子刺激的骨髓来源巨噬细胞产生 12，其表达的主要细胞是基质细胞和脂肪细胞，尤其在人类和啮齿动物皮下的白色脂肪组织中呈高表达 13。人类BIGE数据库中的METRNL表达数据显示，其最高水平的表达在激活的单核细胞中，其次是在消化道、呼吸道黏膜组织和皮肤中，例如在结节性瘙痒、日光性角化病、银屑病和特应性皮炎的患者皮肤中METRNL呈高表达，在人类类风湿关节炎的滑膜液中其表达也明显上调，说明其在先天性和获得性免疫应答中起重要作用 17。Rao等 14发现，METRNL能明显促进白色脂肪中的与褐色脂肪产热及抗炎细胞因子有关的基因表达，促进白色脂肪棕色化,参与炎症反应的调控 15。Jung 等 16 研究发现，METRNL处理可抑制炎症标志物，如抑制核因子kB核易位、抑制NF-kB抑制蛋白磷酸化、抑制炎症性细胞因子IL-6、T NF-和单核细胞趋化蛋白1的表达，减轻炎症反应。METRNL可能以自分泌或旁分泌的方式作用于局部脂肪组织，通过过氧化物酶体增殖物激活受体起到胰岛素增敏作用 17,改善机体的胰岛素抵抗 18。研究表明，肥胖患者血清中METRNL水平较低 19-2 1,本研究结果与该结论一致。Lee等 2 2 发现，METRNL通过钙依赖性AMPK2途径增加骨骼肌细胞的葡萄糖摄取，并以AMPK2依赖的方式增加葡萄糖转运蛋白4转录抑制因子组蛋白脱乙酰酶的磷酸化，表明METRNL蛋白表达上调，可改善全身组织代谢，有效减少脂肪堆积、减轻肥胖 2 3。综上所述,PCOS是一种复杂的内分泌及代谢异常所致疾病，表现为月经不调、不孕、多毛和/或痤疮、肥胖等症状，慢性炎症、IR在其发病机制中起到生殖医学杂志2 0 2 3年9 月第32 卷第9 期关键的作用。METRNL是一种具有多种新功能的讨论新型分泌蛋白，有神经营养活性和抗炎作用，在白色脂肪棕色化、改善脂肪功能、拮抗肥胖诱导的胰岛素抵抗等方面发挥重要作用。本研究结果显示,PCOS患者血清中METRNL表达显著降低，伴有肥胖的PCOS患者血清METRNL表达水平也显著低于正常体重的PCOS患者；伴有 IR的PCOS患者血清METRNL表达水平也显著低于不伴有IR的PCOS患者；所有研究对象中 METRNL与BMI、LH/FSH值、IR、T 均呈负相关，在PCOS患者中METRNL与BMI、I R 均呈负相关。由此我们分析METRNL可能参与PCOS的发生，METRNL可能是一个新的有意义的PCOS的治疗靶点。因此我们是否可以大胆假设,PCOS患者使用METRNL治疗后，将有效控制患者体重，改善IR，缓解炎症状态，进而对PCOS的治疗起到正面效应，当然这还需要我们开展大量的动作实验以及临床实验为此提供理论基础。【参考文献】1 Patel S.Polycystic ovary syndrome(PCOS),an inflammatory,systemic,lifestyle endocrinopathyJJ.J Steroid Biochem MolBiol,2018,182:27-36.2Miao ZW,Hu WJ,Li ZY,et al.Involvement of the secretedprotein METRNL in human diseases JJ.Acta PharmacolSin,2020,41:1525-1530.3 Wang C,Pan Y,Song J,et al.Serum METRNL level iscorrelated with insulin resistance,but not with b-cell functionin type2diabeticsJJ.Med Sci Monit,2019,25:8968-8974.4 Jung TW,Pyun DH,Kim TJ,et al.Meteorin-like protein(METRNL)/IL-41 improves LPS-induced inflammatoryresponses via AMPK or PPARo-mediated signaling 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