穿山龙抗急性痛风性关节炎的肝脏代谢组学研究(1)
[摘要] 该研究通过肝脏代谢组学研究从肝论治痛风的科学性以及穿山龙提取物对急性痛风性关节炎模型大鼠的防治作用，并找到其相关的潜在生物标志物和相关代谢通路。课题组采用尿酸钠(MSU)诱导的急性痛风性关节炎模型并运用UPLC-TOF-MS结合模式识别方法探讨穿山龙提取物干预急性痛风性关节炎的潜在生物标志物及其相关代谢通路。鉴别出了11个共同的潜在生物标志物，其中穿山龙对正常大鼠的潜在干预靶点中，4个上调，4个下调；穿山龙对急性痛风性关节炎模型大鼠的潜在干预靶点中，5个代谢物在MSU诱导下被上调，5个代谢物被下调，而穿山龙提取物表现出纠正磷酸腺苷、5-甲基四氢叶酸、氧化型谷胱甘肽、次黄嘌呤、二十二碳六烯酸、谷胱甘肽、尿苷二磷酸葡萄糖、肌苷这8个代谢物异常表达的趋势，其相关性最强的代谢通路是谷胱甘肽代谢、淀粉与蔗糖代谢和嘌呤代谢。故推测穿山龙可以通过干预急性痛风性关节炎状态下和生理状态下肝脏中内源性代谢产物变化，实现防治急性痛风性关节炎作用。
[关键词] 急性痛风性关节炎； 穿山龙； 尿酸钠； 肝脏代谢组学
[Abstract] To explore the prevention and protection effect of Diosocorea nipponica (DNM)) on acute gouty arthritis (AGA) rats based on liver metabonomics， and find potential biomarkers and related pathways. AGA model rats were induced by monosodium urate crystal suspension. UPLC-TOF-MS coupled with pattern recognition technique was employed to find out the potential biomarkers and related metabolic pathways. Eleven common potential biomarkers were identified. Among the potential intervention targets in normal rats given by DNM， 4 biomarkers were up-regulated， and the other 4 targets were down regulated. Among the potential intervention targets in AGA rats given by DNM， 5 metabolites were up-regulated by MSU and 5 metabolites were down regulated. The abnormal expression levels of adenosine monophosphate， 5-methyltetrahydrofolic acid， oxidized glutathione， hypoxanthine， docosahexaenoic acid， glutathione， uridine diphosphate glucose and inosine could be corrected by DNM extract. Three pathways were founded with greatest correlation， including purine metabolism， starch and sucrose metabolism and glutathione metabolism. Therefore， it could be inferred that D. nipponica has the effect for anti-acute gouty arthritis by intervening endogenous metabolites from the liver under physiological condition and acute gouty arthritis condition.
[Key words] acute gouty arthritis； Diosocorea nipponica； monosodium urate； liver metabonomics
《醫学传灯·痛风》记载：“痛风者，遍身疼痛，昼减夜甚，痛彻筋骨…皆由肝经血少火盛，热极生风，非是外来风邪…”[1]，而现代医学认为痛风的原因是尿酸含量过高，其根源与肝脏代谢异常有关[2]，因此让痛风从肝脏得到根治是个亟待解决的问题，但从肝论治痛风是否具有科学性仍需要临床和实验进一步的检验。穿山龙有祛风除湿、舒筋通络、活血止痛之功，用于各种关节炎[3]。在东北物产丰富，加大对穿山龙的开发，为解除患者疼痛、扩大社会生产力、北药开发战略的实施有极其深远的意义。前期实验表明穿山龙对高尿酸血症及痛风性关节炎有良好的治疗作用[4-9]，基于此运用肝脏代谢组学，预期从整体观探讨穿山龙对生理及急性痛风性关节炎状态下大鼠的影响，寻找穿山龙作用于肝脏中的潜在靶点，验证从肝论治痛风的科学性，模拟出穿山龙抗痛风性关节炎的作用通路及其作用机制。
1 材料
QI软件(Waters 3.0)；UPLC-TOF-MS超高液相色谱-飞行时间-质谱仪(美国Waters公司)；KDC-160HR高速冷冻离心机(科大创新有限公司中佳分公司)；MassLynx V4.1工作站(美国Waters公司)。
乙腈(色谱级，美国Thermo Fisher 科技公司)；甲酸(色谱级，美国Dikma 科技公司)；亮氨酸脑啡肽(美国Sigma-Aldrich公司)；尿酸盐(美国Sigma-Aldrich公司)；蒸馏水(屈臣氏)。黑龙江中医药大学中药毒理学实验室按本课题组的前期研究方法制得穿山龙提取物，其中伪原薯蓣皂苷、原薯蓣皂苷、甲基原薯蓣皂苷的总含量达到 60%[10]。 (刘树民 张宁 于栋华 王宇 周琦 卢芳)
[关键词] 急性痛风性关节炎； 穿山龙； 尿酸钠； 肝脏代谢组学
[Abstract] To explore the prevention and protection effect of Diosocorea nipponica (DNM)) on acute gouty arthritis (AGA) rats based on liver metabonomics， and find potential biomarkers and related pathways. AGA model rats were induced by monosodium urate crystal suspension. UPLC-TOF-MS coupled with pattern recognition technique was employed to find out the potential biomarkers and related metabolic pathways. Eleven common potential biomarkers were identified. Among the potential intervention targets in normal rats given by DNM， 4 biomarkers were up-regulated， and the other 4 targets were down regulated. Among the potential intervention targets in AGA rats given by DNM， 5 metabolites were up-regulated by MSU and 5 metabolites were down regulated. The abnormal expression levels of adenosine monophosphate， 5-methyltetrahydrofolic acid， oxidized glutathione， hypoxanthine， docosahexaenoic acid， glutathione， uridine diphosphate glucose and inosine could be corrected by DNM extract. Three pathways were founded with greatest correlation， including purine metabolism， starch and sucrose metabolism and glutathione metabolism. Therefore， it could be inferred that D. nipponica has the effect for anti-acute gouty arthritis by intervening endogenous metabolites from the liver under physiological condition and acute gouty arthritis condition.
[Key words] acute gouty arthritis； Diosocorea nipponica； monosodium urate； liver metabonomics
《醫学传灯·痛风》记载：“痛风者，遍身疼痛，昼减夜甚，痛彻筋骨…皆由肝经血少火盛，热极生风，非是外来风邪…”[1]，而现代医学认为痛风的原因是尿酸含量过高，其根源与肝脏代谢异常有关[2]，因此让痛风从肝脏得到根治是个亟待解决的问题，但从肝论治痛风是否具有科学性仍需要临床和实验进一步的检验。穿山龙有祛风除湿、舒筋通络、活血止痛之功，用于各种关节炎[3]。在东北物产丰富，加大对穿山龙的开发，为解除患者疼痛、扩大社会生产力、北药开发战略的实施有极其深远的意义。前期实验表明穿山龙对高尿酸血症及痛风性关节炎有良好的治疗作用[4-9]，基于此运用肝脏代谢组学，预期从整体观探讨穿山龙对生理及急性痛风性关节炎状态下大鼠的影响，寻找穿山龙作用于肝脏中的潜在靶点，验证从肝论治痛风的科学性，模拟出穿山龙抗痛风性关节炎的作用通路及其作用机制。
1 材料
QI软件(Waters 3.0)；UPLC-TOF-MS超高液相色谱-飞行时间-质谱仪(美国Waters公司)；KDC-160HR高速冷冻离心机(科大创新有限公司中佳分公司)；MassLynx V4.1工作站(美国Waters公司)。
乙腈(色谱级，美国Thermo Fisher 科技公司)；甲酸(色谱级，美国Dikma 科技公司)；亮氨酸脑啡肽(美国Sigma-Aldrich公司)；尿酸盐(美国Sigma-Aldrich公司)；蒸馏水(屈臣氏)。黑龙江中医药大学中药毒理学实验室按本课题组的前期研究方法制得穿山龙提取物，其中伪原薯蓣皂苷、原薯蓣皂苷、甲基原薯蓣皂苷的总含量达到 60%[10]。 (刘树民 张宁 于栋华 王宇 周琦 卢芳)