2型糖尿病肾病CX3CR1基因多态性相关因素及中医证型分析

Relationship Between the Correlated Factors of CX3CR1 Mutations and the Different TCM Syndromes in Type-2 Diabetic Nephropathy

目的探讨糖尿病肾病(DN)患者CX3CR1-V249I基因突变与中医证型、尿微量白蛋白与肌酐比值(ACR)及炎症因子之间的关系。方法应用聚合酶链反应限制性片段长度多态性的方法及反向测序法检测108例DN患者CX3CR1-V249I多态性情况,同时测定DN患者血浆同型半胱氨酸(Hcy)、糖化血红蛋白(Hb A1c)、ACR、低密度脂蛋白(LDL)、核因子-κB(NF-κB)、白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、趋化因子Fractalkine(FKN)含量,并记录患者的中医证型。结果108例DN患者中CX3CR1基因VV型87例,Ⅵ型17例,Ⅱ型4例。中医证型与CX3CR1-V249I基因型之间具有相关性(χ2=10.136,P<0.05),VV型基因多辨证为气阴两虚型,Ⅵ和Ⅱ型以阴虚燥热为主。与阴虚燥热型和阴阳两虚型比较,气阴两虚型患者具有更高的ACR水平[(948.3±36.4)mg/g比(919.3±31.5)mg/g、(921.8±37.9)mg/g,F=4.946,P<0.05];与痰湿证、湿热证比较,兼血瘀证患者具有更高的ACR水平[(954.3±34.3)mg/g比(921.5±36.9)mg/g、(917.8±37.1)mg/g,F=7.283,P<0.05]。CX3CR1-V249I基因突变与Hcy、ACR水平相关(t=3.601、2.692,P均<0.05)。且V249I野生型(VV型)患者较突变型(Ⅵ型+Ⅱ型)患者NF-Κb[(0.94±0.16)ng/m L比(0.75±0.17)ng/m L,t=7.926,P<0.05]、IL-6[(170.75±34.01)pg/m L比(147.43±27.23)pg/m L,t=5.472,P<0.05]、TNF-α[(95.91±19.21)ng/m L比(81.12±13.50)ng/m L,t=2.887,P<0.05]含量要高,FKN则在突变型患者表达更多[(1.10±0.19)ng/m L比(0.83±0.12)ng/m L,t=9.246,P<0.05)。结论 DN患者CX3CR1-V249I基因多态性与Hcy、ACR、NF-κB、IL-6、TNF-α、FKN及中医证候相关。

Objective To explore the relationship between CX3CR1-V249I mutation and the different syndromes of TCM, albumin to creatinine ratio（ACR） and inflammatory cytokines in patients with type-2 diabetic nephropathy（DN）. Methods By polymerase chain reaction and restriction fragment length polymorphism（PCR-RFLP） and re verse sequencing method, CX3CR1-V249I gene polymorphism was analyzed in 108 patients with DN. The levels of homocysteine（Hcy）, glycosy lated hemoglobin（Hb A1c）, ACR, low-density lipoprotein and the serum contents of NF-κb, IL-6, TNF-α, and FKN were detected. The TCM syndromes of DN in patients were recorded. Results Out of 108 patients, 87 had VV wild genotype and 21 had non-wild genotype including 17 of VI genotype and 4 of homozygote（II genotype）. The CX3CR1-V249I gene polymorphisms was related to the TCM syndromes of DN（χ-2=10.136, P〈0.05）. Patients with genotype VV had mostly the syndrome of Qi and Yin deficiency, and patients with genotype VI＋II had mostly the syndrome of Yin deficiency with heat. Patients with the syndrome of Qi and Yin deficiency had higher ACR level compared with patients with the syndrome of Yin deficiency with heat and the syndrome of Yin and Yang deficiency（948.3±36.4mg/g vs 919.3±31.5mg/g and 921.8±37.9mg/g; F=4.946, P〈0.05）; patients with the syndrome of Qi and Yin deficiency accompanying to blood stasis had higher ACR level compared with patients with the syndrome of Qi and Yin deficiency accompanying to phlegmatic hygrosis or dampheat constitution（954.3±34.3mg/g vs 921.5±36.9mg/g and 917.8±37.1mg/g; F=7.283, P〈0.05）. CX3CR1-V249 I mutation was related with serum Hcy level and ACR level（t=3.601, 2.692, P〈0.05）. The serum contents of NF- κB,IL-6, and TNF-α were increased significantly in the patients with VV genotype than those with VI ＋ II genotypes（NF-κB： 0.94±0.16ng/m L vs 0.75±0.17ng/m L, t=7.926, P〈0.05; IL-6： 170.75±34.01pg/ml vs 147.43±27.23pg/ml, t=5.472, P〈0.05; TNF-α： 95.91±19.21ng/m L vs 81.12±13.50ng/m L, t=2.887, P〈0.05）, but FKN was lower（0.83±0.12ng/m L vs 1.10±0.19ng/m L, t=9.246, P〈0.05）. Conclusion The CX3CR1 polymorphism in patient of type-2 DN was related with serum Hcy, ACR, NF- κB, IL-6, TNF-α, and FKN and TCM syndromes.