湿热内蕴型高尿酸血症复合模型的建立及清利湿热方清利湿热降血尿酸的功效评价

Establishment of a compound model of damp-heat intrinsic hyperuricemia and the evaluation of the efficacy of Qingli Damp-heat Formula for clearing damp-heat and lowering blood uric acid

目的:建立湿热内蕴型高尿酸血症复合模型,探讨清利湿热方降血尿酸作用及清利湿热功效。方法:采用灌胃氧嗪酸钾,并复合饮用蜂蜜水,隔天灌胃猪油脂或酒,建立湿热内蕴型高尿酸血症复合模型。采用全自动生化仪检测大鼠血清尿酸(UA)、尿素氮(BUN)、血肌肝(Scr)水平,酶联免疫法检测血清中黄嘌呤氧化酶(XOD)、脱苷脱氨酶(ADA)、尿酸盐转运体(UAT)、有机阴离子转运体(OAT)含量及热休克蛋白70(HSP70)、肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)水平,以及血清胃泌素(GAS)、血浆胃动素(MTL)含量,免疫组化法检测结肠组织水通道蛋白3(AQP3)、水通道蛋白4(AQP4)表达量,Western Blot测定结肠组织中TLR4、NF-κBp65蛋白表达量。结果:与空白组比较,模型组大鼠血清UA、XOD、ADA、HSP70、TNF-α、IL-1β升高(P<0.01),OAT、GAS降低(P<0.05,P<0.01),结肠组织AQP4含量及TLR4、NF-κBp65蛋白表达量升高(P<0.01,P<0.05)。与模型组比较,清利湿热方低、中、高剂量组能降低UA水平(P<0.01,P<0.05)以及AQP4、HSP70含量(P<0.01,P<0.05),升高GAS水平(P<0.01);清利湿热方低剂量组降低ADA(P<0.05);清利湿热方中剂量组降低XOD、OAT、IL-1β(P<0.01,P<0.05);清利湿热方高剂量组降低Scr(第15天)、ADA、TNF-α、IL-1β(P<0.05,P<0.01)。结论:清利湿热方具有降血尿酸作用及清利湿热功效,为降血尿酸中药产品的中医功效评价研究提供了研究方法及策略,为中药产品的辨证应用、精准应用提供了实验依据。

Objective:To establish a compound model of damp-heat intrinsic hyperuricemia,and to explore the blood uric acid-lowering effect of Qingli Damp-heat Formula and the effect of clearing damp-heat based on the correlation between syndrome and effect.Methods:A hyperuricemia model was established by gavage of potassium oxoxazine,combined with freely drinking honey water,and gavage with oil and liquor every other day to establish a composite model of damp-heat intrinsic hyperuricemia.Using an automatic biochemical analyzer to detect the serum UA,BUN,Scr levels,and ELISA to detect the serum XOD,ADA,UAT,OAT content,HSP70,TNF-α,IL-1βlevel,serum GAS,plasma MTL content,immunohistochemical method was used to detect the expression of AQP3 and AQP4 in colon tissue,and Western Blot was used to detect the expression of TLR4,NF-κBp65 in colon tissue.Results:Compared with the blank group,serum UA,XOD,ADA,HSP70,TNF-αand IL-1βin the model group increased(P<0.01),OAT and GAS decreased(P<0.05,P<0.01),AQP4 content in colon tissue and the expression of protein TLR4 and NF-κBp65 increased(P<0.01,P<0.05).Compared with the model group,the low,medium and high dose groups of Qingli Dampheat Formula groups could reduce the level of UA(P<0.01,P<0.05),and AQP4,HSP70 content(P<0.01,P<0.05),and increased the level of GAS(P<0.01),Qingli Damp-heat Formula low-dose group decreased ADA(P<0.05),Qingli Damp-heat Formula middle-dose group decreased XOD,OAT,IL-1β(P<0.01,P<0.05),Qingli Damp-heat Formula high-dose group decreased Scr(at 15th day),ADA,TNF-α,IL-1β(P<0.05,P<0.01).Conclusion:Qingli Damp-heat Formula have the effect of lowering blood uric acid and clearing damp-heat.The research provides research methods and strategies for the evaluation of TCM efficacy of TCM products for lowering blood uric acid,and provides experimental basis for the application of syndrome differentiation and precise application of TCM products.