基于血清蛋白组学探讨黄芩清热除痹胶囊改善急性痛风性关节炎急性炎症的作用研究

A preliminary study on effect of Huangqin Qingre Chubi Capsule in improving acute inflammation of acute gouty arthritis based on serum proteomics

目的基于血清蛋白组学初步探讨黄芩清热除痹胶囊(Huangqin Qingre Chubi Capsule,HQC)改善急性痛风性关节炎(acute gouty arthritis,AGA)急性炎症的作用研究。方法运用Raybiotech细胞因子抗体芯片筛选治疗前10名AGA患者和10名健康志愿者的血清标本AGA特异性蛋白质。选取3例AGA患者及健康志愿者,采用Western blotting法检测HQC干预前后特异性蛋白质的表达量;选取60例AGA患者及健康志愿者,运用ELISA法检测HQC干预后炎症因子白细胞介素-1β(interleukin-1β,IL-1β)、IL-6、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、转化生长因子-β(transforming growth factor-β,TGF-β)、超敏C-反应蛋白(hypersensitiveC-reactiveprotein,hs-CRP)水平。结果基于蛋白质组学确定了4种AGA特异性蛋白质,包括人肿瘤坏死因子受体超家族成员II(tumor necrosis factor receptor II,TNF-RII)、人巨噬细胞炎性蛋白1β(macrophage inflammatory protein 1β,MIP-1β)、人白细胞介素-8(IL-8)、人粒细胞巨噬细胞集落刺激因子(granulocyte macrophage colony stimulating factor,GM-CSF)。Western blotting检测结果显示对照组与AGA患者治疗前比较,TNF-RII、MIP-1β、IL-8、GM-CSF蛋白水平差异显著,AGA患者治疗前TNF-RII、MIP-1β、IL-8蛋白呈现高表达,GM-CSF蛋白呈现低表达。与治疗前比较,经HQC治疗后AGA患者血清TNF-RII、MIP-1β、IL-8水平显著降低,GM-CSF水平显著升高。ELISA检测结果显示,HQC治疗前后AGA患者血清TNF-RII、MIP-1β、IL-8、GM-CSF水平差异明显,与治疗前比较,治疗后AGA患者血清TNF-RII、MIP-1β、IL-8水平显著降低(P<0.01),GM-CSF水平显著升高(P<0.01)。结论HQC可能是通过调控GM-CSF、IL-8、MIP-1β和TNF-RII等特异性蛋白质的表达,从而改善AGA患者炎症状态,提高机体抗炎能力。

Objective To study the effect of Huangqin Qingre Chubi Capsule(黄芩清热除痹胶囊,HQC)in improving acute inflammation of acute gouty arthritis based on serum proteomics.Methods The Raybiotech cytokine antibody chip was used to screen the AGA-specific proteins in the serum samples of the top 10 AGA patients and 10 normal healthy people.In the study of selecting three clinical patients and normal people,Western blot was used to detect specific proteins.In the study of selecting 60 clinical patients and normal people,the ELISA method was used to analyze the inflammatory parameters after HQC intervention,including interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α),transforming growth factor-β(TGF-β),and high-sensitivity C-reactive protein(hs-CRP)for detection.Results Four AGA-specific proteins were identified based on proteomics,including human tumor necrosis factor receptor superfamily member II(TNF-R II),human macrophage inflammatory protein 1β(MIP-1β),human interleukin-8(IL-8),and human granulocyte macrophage colony stimulating factor(GM-CSF).Western blotting results showed that compared with the control group and the AGA patients before treatment,the protein levels of TNF-RII,MIP-1β,IL-8 and GM-CSF were significantly different;The protein levels of TNF-RII,MIP-1β,IL-8 A were high and GM-CSF protein level was low in AGA patients before treatment.Compared with those before treatment,serum TNF-RII,MIP-1β,IL-8 levels in AGA patients were significantly decreased after treatment,and GM-CSF levels were increased significantly.ELISA test showed that serum TNF-RII,MIP-1β,IL-8 and GM-CSF levels of AGA patients before and after HQC treatment were significantly different.Compared with before treatment,the levels of serum TNF-RII,MIP-1βand IL-8 of AGA patients after treatment were significantly reduced(P<0.01),and the level of GM-CSF was significantly increased(P<0.01).Conclusion The mechanism of HQC improving the inflammatory state of AGA patients may be related to the improvement of specific proteins such as GM-CSF,IL-8,MIP-1βand TNF-RII,thereby improving the inflammatory state of AGA patients and enhancing the anti-inflammatory ability of body.