基于分子对接技术的结果研究藤茶总黄酮抗痛风性关节炎的作用机制

Mechanism of Total Flavonoids from Ampelopsis grossedentata against Gouty Arthritis Based on Molecular Docking Technology

目的:利用分子对接技术结合动物实验对藤茶总黄酮防治痛风性关节炎(gouty arthritis,GA)的作用机制进行研究。方法:利用分子对接技术,将藤茶总黄酮主要药效成分与GA相关治疗靶点腺苷脱氨酶(adenosine deaminase,ADA)、黄嘌呤氧化酶(xanthine oxidase,XOD)、三磷酸腺苷结合盒转运蛋白G2(adenosine triphosphate binding cassette transporter G2,ABCG2)、白细胞介素-1β(interleukin-1β,IL-1β)、IL-6和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)分别进行分子对接模拟。建立高尿酸血症(hyperuricemia,HUA)合并GA大鼠复合模型,对大鼠的步态行为、踝关节炎症指数进行评分,观察踝关节的肿胀程度;检测大鼠血尿酸(UA)水平、XOD和ADA的活性、IL-1β、IL-6和TNF-α的含量;采用免疫组化法测定ABCG2的蛋白表达。结果:分子对接结果显示,藤茶总黄酮关键药效成分二氢杨梅素、杨梅素、芹菜素、槲皮素和橙皮素与各疾病靶点的自由结合能均较高,提示藤茶总黄酮可能是通过作用于ADA、XOD、ABCG2、IL-1β、IL-6和TNF发挥其预防和治疗GA的作用。动物实验结果表明,藤茶总黄酮1、2 g/kg能有效改善GA大鼠异常步态行为、降低大鼠炎症指数、缓解踝关节炎症反应、减小踝关节肿胀度;能显著降低UA含量,抑制XOD、ADA的活性,促进ABCG2的蛋白表达,并显著降低炎症因子IL-1β、IL-6、TNF-α的含量(均P<0.05或P<0.01)。结论:分子对接模拟结果与验证实验结果相一致,均表明藤茶总黄酮抗GA的作用机制可能与降低UA含量,抑制XOD和ADA的活性,促进尿酸盐转运蛋白ABCG2的表达,促进尿酸代谢,并减少炎症因子IL-1β、IL-6和TNF-α的分泌,抑制炎症反应有关。

Objective:To study the mechanism of total flavonoids from Ampelopsis grossedentata(AGTF)in the prevention and treatment of gouty arthritis(GA)by molecular docking technology combined with animal experiments.Methods:Molecular docking simulation was performed between the main pharmacodynamic components of AGTF and GA related therapeutic targets including adenosine deaminase(ADA),xanthine oxidase(XOD),adenosine triphosphate binding cassette transporter G2(ABCG2),interleukin 1β(IL1β),IL6,and tumor necrosis factor-α(TNF-α).A rat model of hyperuricemia(HUA)combined with GA was established.The gait behavior and ankle inflammation index of rats were scored,and the degree of ankle swelling was observed.Uric acid(UA)level,the activities of XOD and ADA,the content of IL1β,IL6,and TNF-αwas detected.The protein expression of ABCG2 was determined by immunohistochemical method.Result:The molecular docking results showed that the key pharmacodynamic components of AGTF including dihydromyricetin,myricetin,apigenin,quercetin,and hesperidin showed high free binding energies with disease-related targets,which suggested that AGTF played its role in the prevention and treatment of GA by acting on ADA,XOD,ABCG2,IL1β,IL6,and TNF.The animal experimental results showed that 1,2 g/kg AGTF effectively improved the abnormal gait behavior of GA rats,reduced the inflammation index,alleviated the ankle inflammation,and reduced the ankle swelling degree of rats.AGTF significantly down-regulated UA level,and inhibited the activities of XOD and ADA.AGTF also promoted the protein expression of ABCG2,and significantly reduced the content of inflammatory factors including IL1β,IL6,and TNF-α(P<0.05 or P<0.01).Conclusion:The molecular docking results are consistent with the experimental results,suggesting that the mechanism of AGTF against GA may be related to the reduction of UA level,inhibition of XOD and ADA activities,promotion of ABCG2 expression and UA metabolism,reduction of the secretion of IL1β,IL6,and TNF-α,and inhibition of inflammatory response.