药学学报

从Wnt/β-catenin串扰轴探究湿热痹颗粒治疗痛风性关节炎湿热证的作用机制

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李欣 , 毛霞 , 马旺明 , 江红 , 付璐 , 向小梅 , 陈文佳 , 李玮婕 , 王萍 , 张彦琼 ^* , 许海玉 ^*

药学学报 | 研究论文 2023,58(8): 2424-2433

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药学学报 |研究论文 2023 , 58 (8) : 2424 -2433

从Wnt/β-catenin串扰轴探究湿热痹颗粒治疗痛风性关节炎湿热证的作用机制

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李欣, 毛霞, 马旺明, 江红, 付璐, 向小梅, 陈文佳, 李玮婕, 王萍, 张彦琼^*, 许海玉^*

作者信息

中国中医科学院中药研究所, 北京 100700

通讯作者:

*张彦琼, E-mail: yqzhang@icmm.ac.cn;

许海玉, Tel: 86-10-64014411, E-mail: hyxu@icmm.ac.cn

Investigation on the pharmacological mechanisms of Shirebi granules in treating gouty arthritis with damp-heat syndrome based on crosstalk of Wnt/β-catenin signaling

Xin LI, Xia MAO, Wang-ming MA, Hong JIANG, Lu FU, Xiao-mei XIANG, Wen-jia CHEN, Wei-jie LI, Ping WANG, Yan-qiong ZHANG^*, Hai-yu XU^*

Affiliations

Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China

出版时间: 2023-08-12 doi: 10.16438/j.0513-4870.2023-0540

文章导航

摘要

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痛风性关节炎是一类由自身免疫异常引起的代谢性风湿病, 目前临床中用于治疗此类疾病的一线西药易引起不良反应, 从中医药中寻求治疗药物成为研究热点。中医临床将痛风性关节炎归为湿热痹证, 对症治疗的湿热痹颗粒疗效确切, 但作用机制尚不清晰。本研究旨在探究湿热痹颗粒治疗痛风性关节炎湿热证的作用机制。首先, 建立“湿热痹方剂候选靶标-痛风性关节炎湿热证相关基因”的相互作用网络, 通过计算网络节点拓扑特征值, 筛选湿热痹颗粒治疗痛风性关节炎湿热证的关键候选靶标, 并对关键候选靶标进行功能挖掘, 发现湿热痹颗粒候选靶标可通过调节Wnt/β-catenin串扰轴, 减轻炎症反应和干预脂质代谢生物过程, 从而发挥其缓解痛风性关节炎湿热证的作用。进一步基于病证结合大鼠模型的实验验证表明, 湿热痹颗粒可显著改善痛风性关节炎湿热证大鼠的关节炎严重程度, 降低关节炎症与步态评分、肿胀指数, 升高机械痛阈值(P < 0.05), 降低痛风性关节炎湿热证大鼠血清炎症因子IL-1β、IL-6、TNF-α的含量(P < 0.01); 还能显著降低痛风性关节炎湿热证大鼠关节局部温度, 提高热痛阈值, 增加饮水量(P < 0.01), 抑制脂肪酸氧化相关酶含量的异常变化(P < 0.01); Western blot机制层面的验证结果显示, 湿热痹颗粒可升高痛风性关节炎湿热证大鼠Wnt、β-catenin蛋白表达水平(P < 0.01), 降低p65、p-p65、PPARγ蛋白表达(P < 0.01), 即湿热痹颗粒可能通过调节Wnt/β-catenin串扰轴矫正“炎症-免疫”失衡和脂质代谢紊乱, 发挥缓解疾病严重度的作用。本研究为中药病-证-方复杂关联机制的解析提供方法学参考, 为中成药优势品种的临床合理用药及痛风性关节炎的新药研发奠定了良好基础。本研究获得中国中医科学院医学实验中心实验动物伦理委员会批准(批准号: ERCCACMS11-2302-08)。

关键词

湿热痹颗粒 / 痛风性关节炎 / 湿热证 / "免疫-炎症"失衡 / 脂质代谢

Abstract

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Gouty arthritis is a type of metabolic rheumatic disease caused by autoimmune abnormalities. Currently, the use of Western medicine in the clinical treatment of gouty arthritis has been associated with a high risk of adverse reactions. Therefore, there is a growing interest in exploring therapeutic drugs from traditional Chinese medicine as a potential alternative. According to the theory of traditional Chinese medicine, gouty arthritis has been classified as damp-heat arthralgia syndrome. Shirebi granules has been found to have good clinical efficacy in treating gouty arthritis. However, its underlying pharmacological mechanisms remain unclear. To address this problem, the study first established the interaction network of candidate targets for Shirebi granules, which is used to treat damp-heat syndrome of gouty arthritis. Then, the key candidate targets of Shirebi granules for treating gouty arthritis with damp-heat syndrome were screened by calculating the topological features of the network nodes. Then, the functional mining of the key candidate targets revealed that the candidate targets of Shirebi granules may intervene in the biological process of inflammatory response and lipid metabolism through the crosstalk of Wnt/β-catenin signaling. To verify the effectiveness of Shirebi granules in treating gouty arthritis with damp-heat syndrome, a rat model was established. The results demonstrated that the granules significantly improved the severity of arthritis in rats with this condition, reduced joint inflammation, gait score, swelling index, increased mechanical pain threshold (P < 0.05), and reduced the content of serum inflammatory factors IL-1β, IL-6, and TNF-α in gouty arthritis rats with damp-heat syndrome (P < 0.01) gouty. It was also found that Shirebi granules effectively alleviated the symptoms of dampness heat syndrome such as local joint fever and dry mouth by reducing the temperature of the joints in acute gouty arthritis with damp-heat syndrome (AD) rats, increasing the threshold of heat pain, increasing water intake (P < 0.01), and inhibiting abnormal changes in the content of fatty acid oxidation related enzymes (P < 0.01). Western blot analysis showed that Shirebi granules increased the protein expression levels of Wnt and β-catenin (P < 0.01) while decreasing the protein expression of p65, p-p65 and PPARγ (P < 0.01) in rats with gouty arthritis and damp-heat syndrome. The results showed that Shirebi granules may reverse the "inflammation-immune" imbalance and lipid metabolism disorder by regulating the crosstalk of Wnt/β-catenin signaling, and play a role in alleviating the severity of the disease. This study provides a methodological reference for elucidating the pharmacological mechanisms of traditional Chinese medicine formulas. It also presents research ideas for the appropriate clinical use of Chinese patent medicines and the development of new clinical drugs for gouty arthritis therapy. The animal welfare and experiment procedures of this study were performed in accordance with the regulations of the Experimental Animal Ethics Committee of Experimental Research Center, China Academy of Chinese Medical Sciences (grant No. ERCCACMS11-2302-08).

Key words

Shirebi granules / gouty arthritis / damp-heat syndrome / imbalance of immunity-inflammation / lipid metabolism

引用本文

李欣, 毛霞, 马旺明, 江红, 付璐, 向小梅, 陈文佳, 李玮婕, 王萍, 张彦琼, 许海玉. 从Wnt/β-catenin串扰轴探究湿热痹颗粒治疗痛风性关节炎湿热证的作用机制. 药学学报, 2023 , 58 (8) : 2424 -2433 . DOI: 10.16438/j.0513-4870.2023-0540

Xin LI, Xia MAO, Wang-ming MA, Hong JIANG, Lu FU, Xiao-mei XIANG, Wen-jia CHEN, Wei-jie LI, Ping WANG, Yan-qiong ZHANG, Hai-yu XU. Investigation on the pharmacological mechanisms of Shirebi granules in treating gouty arthritis with damp-heat syndrome based on crosstalk of Wnt/β-catenin signaling[J]. Acta Pharmaceutica Sinica, 2023 , 58 (8) : 2424 -2433 . DOI: 10.16438/j.0513-4870.2023-0540

正文

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痛风性关节炎简称为痛风, 是由血液中过饱和的尿酸以晶体形式析出, 沉积于关节囊、软骨等组织, 从而引起自身免疫的代谢性风湿病^[1]。据《2021中国高尿酸及痛风趋势白皮书》统计, 我国痛风总体发病率为1.1%, 呈明显上升和年轻化趋势^[2]。痛风性关节炎临床上分为无症状期、急性期、间歇期与慢性期4个病程阶段, 其中急性期以发病率高、易于诊断、疼痛剧烈、难以根治等显著特点, 成为痛风性关节炎治疗的关键期之一^[3]。目前, 临床上常用非甾体类抗炎药、秋水仙碱、皮质类固醇等作为急性痛风性关节炎的一线用药, 旨在快速缓解疼痛、减轻关节局部的炎症反应, 疗效显著, 但同时也会引起较为严重的不良反应^[4], 如胃肠道溃疡、出血、慢性肾功能不全等^{[5, 6]}, 而从中医药中寻求更加安全、有效的治疗痛风性关节炎的药物也是研究热点之一。

中医将痛风性关节炎归属于“痹病”, 其流行病学研究表明, 痛风性关节炎急性期以湿热蕴结证为主^[7]。而由经典名方四妙丸化裁而来的湿热痹颗粒是治疗湿热证的代表性中成药, 组方中包含苍术、忍冬藤、地龙、连翘、黄柏、薏苡仁、防风、川牛膝、粉萆薢、桑枝、防己、威灵仙12味中药, 具有祛风除湿、清热消肿和通络定痛的功效。前期临床研究表明, 湿热痹颗粒可有效降低痛风性关节炎患者尿酸水平, 降低炎症因子水平, 临床有效率达95.56%^[8]; 且对痛风性关节炎关节疼痛、关节功能障碍、关节肿胀等临床症状有较好的缓解作用^{[8, 9]}; 相较于单纯西药治疗, 湿热痹颗粒在降低痛风性关节炎患者血尿酸和炎症因子水平及提高患者心理评分等方面有独特优势^[9]。但目前关于湿热痹颗粒在痛风性关节炎方面的报道仅限于药效, 缺乏对其机制的研究, 给品种临床精准定位带来了困难, 往往限制了该品种临床中的应用。因此, 本研究利用痛风性关节炎湿热证(acute gouty arthritis with damp-heat syndrome, AD) 大鼠模型, 在病证方网络关联分析的基础上, 考察湿热痹颗粒治疗痛风性关节炎湿热证的药效, 并初步揭示其作用机制, 为促进该品种临床合理、精准应用提供参考依据。

材料与方法

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实验动物 雄性SPF级大鼠, 体重180 ± 20 g, 6~8周龄, 购自北京华阜康生物科技股份有限公司, 动物许可证: SCXK (京) 2020-0004。由中国中医科学院医学实验中心饲养、造模与观察。实验期间动物自由饮食和进水, 昼夜节律正常。本研究获得中国中医科学院医学实验中心实验动物伦理委员会批准(批准号: ERCCACMS11-2302-08)。

药物与试剂 湿热痹颗粒由苍术、忍冬藤、地龙、连翘、黄柏、薏苡仁、防风、川牛膝、粉萆薢、桑枝、防己、威灵仙12味中药组成。湿热痹颗粒冻干粉制备方法: 苍术、连翘、防风提取挥发油, 剩余药渣弃去, 挥发油与药液备用。取地龙、黄柏等九味, 加水煎煮两次, 二次提取液与提取挥发油药液合并, 滤过, 滤液浓缩成流浸膏, 加3倍乙醇, 静置12 h, 取上清液浓缩至相对密度1.33~1.35 (50 ℃测量) 的清膏, 并冻干成粉末。

秋水仙碱(每片0.5 mg, 云南植物药业有限公司, 国药准字H53020166); 尿酸钠盐(U2875, 美国Sigma-Aldrich公司); 白介素(interleukin, IL) 1β酶联免疫吸附法(enzyme linked immunosorbent assay, ELISA) 试剂盒(ml028514)、IL-6 ELISA试剂盒(ml102828)、肿瘤坏死因子α (tumor necrosis factor alpha, TNF-α) ELISA试剂盒(ml059055)、乙酰辅酶A羧化酶(acetyl-CoA carboxylase, ACC) ELISA试剂盒(ml059228)、肉碱棕榈酰转移酶1 (carnitine palmitoyltransferase 1, CPT-1) ELISA试剂盒(ml059355) 购自上海酶联生物科技有限公司; 脂肪酸β氧化酶(fatty acid β oxidase, FaβO) ELISA试剂盒(CB12027-Ra, 上海科艾博生物技术有限公司); NF-κB p65抗体(A19653)、NF-κB p-p65抗体(AP0475) 购自武汉爱博泰克生物科技有限公司; PPARγ抗体(sc-271392)、Wnt-4抗体(sc-376279) 购自美国Santa Cruz Biotechnology公司。

仪器 Von frey hair疼痛测试仪(美国Stoeltingco公司); RB-200智能热板仪(成都泰盟有限公司); Tanon 5200化学发光成像系统(上海天能科技有限公司); TESTO-875红外热像仪(德图仪器国际贸易有限公司); 游标卡尺(宁波得力工具有限公司); 旋涡混合器(KA-1000北京金北德工贸有限公司)。

湿热痹颗粒化学成分及候选靶标谱的收集和整理 利用ETCM v2.0数据库(http://www.tcmip.cn/ETCM2/front/#/browse/herb)^[10], 结合文献调研, 收集湿热痹颗粒的活性成分(QED ≥ 0.49); 同时通过ETCM v2.0和TCMIP v2.0 (http://www.tcmip.cn/TCMIP/index.php/Home)^[11]平台, 预测成分所对应的靶点(可信度得分 > 0.8)。

痛风性关节炎相关基因集的收集和整理 在MalaCards (https://www.malacards.org/)^[12]、ETCM (https://www.malacards.org/)^[11]、GeneCards (https://www.genecards.org/)^[13]、NCBI (https://www.ncbi.nlm.nih.gov/)^[14]数据库中检索“gouty arthritis”, 并选择“Homo sapiens”, 收集数据库中痛风性关节炎的疾病基因; 同时, 在PubMed中检索主题词“gouty arthritis”, 收集文献中痛风性关节炎的疾病基因; 除此之外, 通过基因表达综合数据库(Gene Expression Omnibus, GEO)^[15]检索“gouty arthritis”, 选择与痛风性关节炎相关的小鼠踝关节测序数据集GSE190138, 并以P < 0.05及log₂|FC| > 1.5为标准筛选差异基因, 同样作为痛风性关节炎的疾病基因集的来源之一。

湿热证相关基因集的收集和整理 CNKI检索“湿热”、“痛风性关节炎”和“临床症状”, 收集痛风性关节炎湿热证特征主症与次症, 并在SoFDA (http://www.tcmip.cn/Syndrome/front/#/)^[16]和Symmap (http://www.symmap.org/)^{[17, 18]}数据库中收集痛风性关节炎湿热证主症与次症基因, 作为痛风性关节炎湿热证的证候基因集。

病-证-方关联网络构建与分析 基于String数据库(version 11.5, https://cn.string-db.org/)^[19]构建“湿热痹颗粒候选靶标-痛风性关节炎湿热证相关基因”生物分子网络, 并计算互作网络的拓扑特征值(连接度、介度、紧密度), 筛选三者均大于中位数的节点作为关键网络靶标。

GO分析和KEGG通路富集分析 利用DAVID数据库(https://david.ncifcrf.gov/)^[20]对关键网络靶标进行GO生物学过程分析和KEGG信号通路富集分析。

痛风性关节炎湿热证动物模型的建立 参照Coderre的经典方法^[21]结合文献调研^[22], 制备痛风性关节炎湿热证模型。除正常组外, 其余6组大鼠均于第13天在右后踝关节处注射50 mg·mL^-1尿酸钠结晶(monosodium urate, MSU) 混悬液100 μL, 对照组大鼠右踝关节则注入等量无菌氯化钠溶液, 以关节囊对侧鼓起为注入标准。同时, 模拟痛风性关节炎的湿热病机, 给予内湿形成的条件刺激(以油脂和52%红星二锅头酒交替灌胃10天), 另施加外界湿热环境刺激(相对湿度约95%, 温度32 ± 2 ℃, 连续15天), 造成痛风性关节炎湿热证模型。

分组及给药 36只大鼠, 随机分为6组, 每组6只: 对照组(control, Con)、痛风性关节炎湿热证模型组(AD)、湿热痹颗粒低剂量组(ADL)、湿热痹颗粒中剂量组(ADM)、湿热痹颗粒高剂量组(ADH) 和秋水仙碱组(Col)。湿热痹颗粒低、中、高剂量为1.25、2.5、5 g·kg^-1, 分别相当于0.5、1、2倍临床等效剂量, 秋水仙碱按临床等效剂量给药, 为0.3 mg·kg^-1。从第1天开始给药, 给药组按照10 mL·kg^-1灌胃, 正常组和模型组灌服等体积蒸馏水, 连续给药15天。

关节炎评价方法 踝关节注射MSU后24 h, 分别从踝关节功能障碍指数^[23]、踝关节炎症指数^{[24, 25]}和踝关节肿胀指数^[26]3个方面对痛风性关节炎进行评价。踝关节功能障碍指数分级标准: 0级, 行走正常; 1级, 下肢微弯、跛行; 2级, 下肢仅接触地面、中度跛行; 3级, 三足行走, 跛行严重。踝关节炎症指数分级标准: 0级, 关节正常; 1级, 骨标志明显、关节轻度红肿; 2级, 骨标志不明显、关节红肿明显; 3级, 出现关节外红肿现象。踝关节肿胀指数测定: 使用数字卡尺测量大鼠右踝关节周长和右踝关节周长, 最小精度为0.01 mm。关节肿胀指数= (右踝关节周长-左踝关节周长)/左踝关节周长。每组重复3次, 取平均值。

疼痛测定方法

机械痛检测方法^[27] 通过Von frey hair机械疼痛测试仪检测大鼠机械痛。分别于第12和14天测量, 每次测量维持3个周期, 中间间隔5 min, 最后计算取平均值。

热痛检测方法^[28] 通过YLS-6B智能热板仪检测大鼠热痛。第12天将大鼠置于(55 ± 0.5) ℃的热板仪, 以大鼠接触热板开始计时, 每组筛选3只在5~30 s内出现舔后足反应的大鼠进行后续实验。于第14天, 将筛选出的大鼠置于热板, 记录大鼠舔后足反应的时间, 每只大鼠测定2次, 每次间隔10 min, 取其平均值即为大鼠的热痛阈值。

饮水量测定 每天开始前测量水瓶中水的体积, 同时记录下开始时间, 每天结束时再次测量水瓶中水的体积, 并记录下结束时间, 饮水量= (开始体积-结束体积)/间隔时间×24 h。

酶联免疫吸附测定法检测IL-1β、TNF-α、IL-6、CPT-1、ACC、FaβO蛋白含量 采用ELISA测定炎症因子及脂肪酸氧化关键酶。按照试剂盒说明书的步骤, 测定大鼠血清中的IL-1β、TNF-α、IL-6、FaβO的含量及肝脏组织中CPT-1、ACC的含量。

免疫印迹法检测踝关节中Wnt、β-catenin、PPARγ、NF-κB蛋白表达 取0.1 g踝关节组织, 加入1 mL含有蛋白酶抑制剂、磷酸酶抑制剂和苯甲基磺酰氟的裂解液, 充分匀浆, 4 ℃、12 000 r·min^-1离心15 min, 取上清。BCA蛋白定量。用7.5% SDS-PAGE凝胶电泳分离蛋白, 并将蛋白转移至PVDF膜上。5%脱脂奶粉室温封闭1.5 h后加入适当浓度的一抗Wnt (1∶500)、β-catenin (1∶1 000)、PPARγ (1∶1 000)、NF-κB p65 (1∶1 000)、NF-κB p-p65 (1∶1 000), 4 ℃孵育过夜。洗膜3次后加入对应二抗, 室温孵育1 h, 再次洗膜3次后, ECL显色并曝光拍照。

统计学分析 利用SPSS 26.0进行统计学分析, 数据均以x ± s表示。采用单因素方差分析进行组间比较, P < 0.05表示差异具有统计学意义。

结果

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1 湿热痹颗粒可能通过调控Wnt/β-catenin串扰轴, 而减轻痛风性关节炎湿热证的炎症反应和干预其脂质代谢环节

数据库检索结合文献调研的方式, 收集到痛风性关节炎湿热证1 215个疾病基因集和744个证候基因集, 并通过ETCM v2.0和TCMIP v2.0获得1 045个湿热痹颗粒方剂靶标谱。通过String数据库提取湿热痹方剂候选靶标和痛风性关节炎湿热证病证相关基因间的相互作用关系, 构建“湿热痹方剂候选靶标-痛风性关节炎湿热证相关基因”相互作用网络, 并计算上述网络节点的拓扑特征值, 通过计算网络中包括连接度、介度和紧密度, 选择三者均大于相应中位数的关键网络靶标, 共筛选出290个关键网络节点, 其中有165个为湿热痹方剂候选靶标。对关键靶标进行网络富集分析, 结果表明湿热痹颗粒方剂靶标显著富集于炎症反应相关通路, 如NF-κB信号通路、Toll样受体信号通路、TNF信号通路、Wnt信号通路等; 此外, 还与脂质代谢过程相关, 如脂肪细胞因子(adipocytokine)、脂肪细胞脂解(lipolysis in adipocytes) 和脂肪发育与代谢(Wnt) 等信号通路(图 1A)。而Wnt/β-catenin信号通路既可调控机体炎症反应, 又是脂质代谢的关键通路, 且通路中包含CREBBP、MAP3K7、GSK3B、PPARD等在内的11个湿热痹颗粒方剂核心靶标(图 1B)。推测湿热痹颗粒可能通过Wnt/β-catenin与NF-κB串扰轴, 从而调节痛风性关节炎湿热证的炎症反应; 通过Wnt/β-catenin与PPAR串扰轴, 从而改善痛风性关节炎湿热证的脂肪代谢。

2 湿热痹颗粒可显著缓解痛风性关节炎湿热证大鼠疾病严重程度

2.1 湿热痹颗粒可显著改善痛风性关节炎湿热证大鼠关节炎宏观评价指标

本研究成功构建了AD大鼠模型, AD模型组大鼠发病率100%, 造模1周后, 大鼠表现出明显的精神倦怠、便溏、活动量减少等现象。与对照组相比, AD模型组大鼠体重增长缓慢, 关节炎症指数、关节功能障碍指数、关节肿胀指数均显著升高(P < 0.001), 机械痛阈值降低(P < 0.001); 与AD模型组相比, 湿热痹颗粒低、中、高剂量可不同程度地升高体重(图 2A)、降低关节炎症指数(P < 0.05, 图 2B)、功能障碍指数(P < 0.001, 图 2C) 与肿胀指数(P < 0.01, 图 2D), 升高机械痛(P < 0.05, 图 2E)。其中, 高剂量组的关节功能障碍指数、机械痛阈值与秋水仙碱组无统计学差异, 但高剂量组对关节肿胀指数的改善效果优于秋水仙碱组(P < 0.001)。

2.2 湿热痹颗粒可显著降低痛风性关节炎湿热证大鼠血清炎性因子水平

与对照组相比, AD模型组大鼠血清的炎症因子IL-6、IL-1β、TNF-α水平显著升高(P < 0.001); 与AD模型组相比, 湿热痹颗粒低、中、高剂量可不同程度地降低血清中IL-6 (P < 0.01, 图 3A)、IL-1β (P < 0.001, 图 3B) 和TNF-α (P < 0.001, 图 3C) 表达水平, 中剂量组药效与秋水仙碱相比无显著差异。

3 湿热痹颗粒具有缓解痛风性关节炎湿热证大鼠湿热证的作用

3.1 湿热痹颗粒可有效改善痛风性关节炎湿热证大鼠口干、发热表征

与对照组相比, AD模型组大鼠饮水量升高, 关节表面温度显著升高(P < 0.001), 热痛阈值明显降低(P < 0.001); 与AD模型组相比, 湿热痹颗粒中、高剂量显著改善其口干、发热等症状, 具体表现为降低AD模型组大鼠饮水量(图 4A)、关节表面温度(P < 0.001, 图 4B), 提高热痛阈值(P < 0.01, 图 4C), 对热痛阈的改善效果与阳性对照药秋水仙碱相比无显著差异。

3.2 湿热痹颗粒可调节痛风性关节炎湿热证大鼠脂肪酸氧化关键酶含量, 抑制脂肪酸氧化

与对照组相比, AD模型组大鼠中脂肪酸氧化关键酶ACC含量显著降低(P < 0.001), CTP-1、FaβO含量显著升高(P < 0.001); 与AD模型组相比, 湿热痹颗粒中、高剂量可不同程度地升高ACC (P < 0.01, 图 5A) 的表达水平及降低CTP-1 (P < 0.001, 图 5B)、FaβO (P < 0.001, 图 5C) 的含量, 中、高剂量组对CTP-1和FaβO的抑制效果与秋水仙碱相比无显著差异。

4 湿热痹颗粒可通过调控Wnt/β-catenin串扰轴相关蛋白表达, 改善痛风性关节炎大鼠“炎症-免疫”失衡与脂质代谢紊乱

与对照组相比, AD模型组大鼠中Wnt、β-catenin蛋白表达水平显著降低(P < 0.01, 图 6A), PPARγ、p65和p-p65蛋白表达水平显著升高(P < 0.001, 图 6B、C); 与AD模型组相比, 湿热痹颗粒中、高剂量可不同程度地升高Wnt (P < 0.001, 图 6A、D)、β-catenin (P < 0.01, 图 6A、E) 表达水平, 下调PPARγ (P < 0.01, 图 6B、F)、p65 (P < 0.001, 图 6C、G)、p-p65 (P < 0.01, 图 6C、H) 蛋白的异常表达, 且高剂量组药效与阳性对照药秋水仙碱相比无显著差异。

讨论

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痛风性关节炎因“其疾昼静而夜发, 即彻骨髓酸疼, 其痛如虎之啮”, 故而又被称为“白虎历节”^[29]。传统医家认为痛风发生“多因体内正气不足, ……湿热痰瘀等病理产物聚于体内, 留滞经络; 又因外邪侵袭内外合邪, 气血凝结不通所致”^[30]。其中, 急性期以湿热蕴结证痛风性关节炎最多见, 约占痛风性关节炎的60%, 临床上常以清热利湿为治则^[31]。湿热痹颗粒中苍术、薏苡仁有健脾祛湿的功效; 忍冬藤、连翘、黄柏可清热泻火, 疏风通络; 防己、粉萆薢可利水祛湿; 地龙、防风、威灵仙、桑枝、川牛膝是祛风活络、通利关节、除湿止痛的常用中药, 全方配伍精妙, 共奏清热利湿、祛风通络、消肿止痛之功效^[8]。

本研究采用油脂、白酒灌胃, 模拟湿热证“脾失健运, 湿浊内生”的病因病机, 联合高温高湿的外界环境, 内外相引、共生湿热^[32]。此方法造模1周后, 模型组大鼠出现便溏、活动次数减少、体重增长缓慢等表现, 符合湿热证“倦怠乏力、大便黏滞不畅、身重困倦、食少纳呆”的临床症状^[33]。“人工气候箱+饮食+单钠尿酸盐”建立的痛风性关节炎湿热证既能体现出痛风性关节炎关节疼痛、肿胀、关节功能障碍的关节炎特点, 又有关节发热、倦怠乏力、口干、便溏等湿热证表型, 与湿热蕴结型痛风性关节炎患者的临床症状相契合^[34]。本实验结果也显示, 相较于痛风性关节炎湿热证模型组, 湿热痹颗粒治疗后, 各剂量组大鼠炎症指数、功能障碍指数、肿胀指数、关节表面温度更低, 机械痛阈值、热痛阈值更高, 且炎症因子水平下降, 脂肪酸氧化相关酶含量异常的情况也被逆转, 表明湿热痹颗粒能有效改善痛风性关节炎湿热证大鼠关节红、肿、热、痛的异常, 缓解关节不利的症状, 调整疼痛敏化的反应, 进一步证明湿热痹颗粒具有治疗痛风性关节炎湿热证的潜力。

本研究依据网络拓扑特征值筛选出湿热痹颗粒治疗痛风性关节炎的关键候选靶标, 并对其功能富集分析发现, 湿热痹颗粒治疗痛风性关节炎湿热证的主要作用环节是调节免疫-炎症失衡与改善脂质代谢紊乱, 与现有的研究报道一致^{[32, 35, 36]}。痛风性关节炎发病机制之一就是尿酸盐沉积引起机体内的炎症反应^{[1, 2]}, 痛风性关节炎患者存在严重的“免疫-炎症”失衡现象; 而组学研究证明湿热证中“水湿内停、痰浊内生”的生物学基础之一可能为甘油代谢障碍^[37]; 临床横断面研究也报道脂代谢异常是痛风性关节炎诱发因素之一^[38]; 湿热证的特征之一即为脂代谢紊乱^[39]。可见“免疫-炎症”失衡与脂质代谢紊乱是痛风性关节炎湿热证的两个重要病理环节, 而上述两个环节可被Wnt/β-catenin串扰。Wnt通路是真核生物中广泛存在的一条经典的、发育中保守的信号传导途径, 在胚胎发育、细胞分化、脂肪代谢、骨发育重建和免疫调控等生物过程中发挥着极其重要的作用^[40-42]。大量研究表明, NF-κB信号传导与典型Wnt通路之间存在物理与生物功能的串扰, 且这种串扰作用进一步调节了炎症相关疾病的发展^{[43, 44]}。一方面, Wnt通路下游的关键功能效应分子β-catenin可与RelA和p50复合^[45], 降低NF-κB信号传导的活性, 还可通过与PI3K相互作用, 抑制NF-κB的功能活性^[46], 从而抑制炎症; 反之, 若β-catenin蛋白被抑制, 会导致NF-κB信号通路的激活^{[47, 48]}。本研究同样发现, 痛风性关节炎湿热证大鼠中β-catenin蛋白表达水平降低, 而NF-κB蛋白表达水平升高, 炎症因子含量也随之上升, 湿热痹颗粒给药后两者蛋白表达水平均得到了改善, 炎症因子含量也显著降低, 结果与文献^[9]相符。另一方面, 有研究报道, NF-κB也可调节Wnt/β-catenin信号通路的活性, 如在小鼠软骨细胞中, NF-κB的激活通过诱导Wnt/β-catenin信号通路中转录因子Lef1的表达, 间接调节β-catenin的转录活性^[49]。总之, Wnt/β-catenin信号通路可通过抑制或激活NF-κB通路从而发挥抗炎或促炎作用, 而NF-κB通路也可正向或负向调节Wnt/β-catenin信号通路, 两种途径的不同组成部分参与这种串扰, 形成了一个复杂的调控网络, 从而在疾病中发挥相应功能^[43]。此外, Wnt信号通路还可通过调控脂肪细胞的分化与发育, 进而调节脂质代谢^{[50, 51]}。有研究报道, Wnt信号通路可与PPARγ相互作用, 调控脂肪细胞分化^[52]。抑制Wnt/β-catenin信号通路, 可促进PPARγ蛋白表达^[53], PPARγ蛋白可诱导脂肪酸氧化酶系与脂蛋白脂酶等, 从而促进脂质的氧化代谢^[54]。本实验结果表明, 痛风性关节炎湿热证大鼠中Wnt、β-catenin蛋白表达水平降低, 而PPARγ蛋白表达升高, 脂肪酸氧化酶系也随之改变, 脂肪酸氧化增强, 湿热痹颗粒给药可有效抑制痛风性关节炎湿热证大鼠中的脂肪酸氧化程度。因此, 本研究揭示了湿热痹颗粒对痛风性关节炎湿热证大鼠关节炎严重程度及湿热证均有明显的改善作用, 并通过调节Wnt/β-catenin串扰轴, 降低NF-κB与PPARγ蛋白表达, 进而降低炎症因子含量与抑制脂肪酸氧化进程, 从而矫正其痛风性关节炎湿热证大鼠体内“免疫-炎症”失衡与脂质代谢紊乱的病理环节, 有效缓解了痛风性关节炎湿热证大鼠关节肿胀、关节不利、发热、口干等症状。

综上, 本研究采用病证方关联网络分析和实验验证手段相结合的策略, 揭示了湿热痹颗粒可有效改善痛风性关节炎湿热证大鼠的疾病严重程度与缓解湿热证症状, 其机制与Wnt/β-catenin串扰轴相关。本研究为中药病-证-方复杂关联机制的解析提供方法学参考, 为中成药优势品种的临床合理用药及痛风性关节炎的新药研发奠定了良好基础。

致谢: 感谢辽宁上药好护士药业(集团) 有限公司提供经费和实验样品的支持。

作者贡献: 李欣参与实验研究、论文撰写及论文修改; 毛霞参与实验指导及论文修改; 马旺明、江红、付璐和向小梅参与动物实验; 陈文佳和李玮婕参与网络药理学分析; 王萍参与实验设计; 张彦琼和许海玉是本文的通讯作者, 负责实验设计及把关、提供本文思路和参与稿件修改等工作。

利益冲突: 本文不存在任何利益冲突。

基金

收起

国家自然科学基金青年科学基金项目(82204711)
中央本级重大增减支项目(2060302)
青年岐黄学者培养项目(国中医药人教函[2022]256号)

参考文献

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文献

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参考文献引证文献

排序方式：

[1]

Dalbeth N, Gosling AL, Gaffo A, et al Gout[J]. Lancet, 2021, 397: 1843-1855.

https://doi.org/10.1016/S0140-6736(21)00569-9

[2]

Wang WF, Zhu F, Jiang YQ Urine pH value combined with serum creactive protein in prediction of acute gout attack in patients with cerebr-al infarction[J]. Zhejiang Med J (浙江医学), 2022, 44: 2294-2297.

[3]

Zeng XJ Interpretation of the Chinese guideline for the management of gout in 2010[J]. Chin J Pract Int Med (中国实用内科杂志), 2012, 32: 438-441.

[4]

Dalbeth N, Choi HK, Joosten LAB, et al Gout[J]. Nat Rev Dis Primers, 2019, 5: 69.

https://doi.org/10.1038/s41572-019-0115-y

[5]

Fu HT, Zhang M, Ci XY, et al Research progress of therapeutic agents for gout and hyperuricemia[J]. Drug Eval Res (药物评价研究), 2021, 44: 1811-1816.

[6]

Yu YY, Xu Y, Zhu XX, et al Therapy and choice of the drugs for acute gouty arthritis[J]. Shanghai Med Pharm J (上海医药), 2015, 36: 14-18.

[7]

Feng DH, Zhao YF, Zhao MY, et al Study on clinical characteristics of patients with acute gouty arthritis based on data mining in the real world[J]. Mod Tradit Chin Med Mater Med World Sci Technol (世界科学技术-中医药现代化), 2020, 22: 3996-4000.

[8]

Han XF, Du TX, Li WY Clinical observation on the treatment of damp heat Bi obstructive rheumatology with Shirebi granules[J]. J Tradit Chin Orthop Traumatol (中医正骨), 2022, 14: 9-11.

[9]

Liu RP, Liu DM, Zeng FX Effect of Shirebi granules assisted with benzbromarone tablets on serum immune and inflammatory indicators in patients with gouty arthritis[J]. World J Integr Tradit West Med (世界中西医结合杂志), 2021, 16: 748-752.

[10]

Zhang YQ, Li X, Shi Y, et al ETCM v2.0: an update with comprehensive resource and rich annotations for traditional Chinese medicine[J]. Acta Pharm Sin B, 2023, 13: 2559-2571.

https://doi.org/10.1016/j.apsb.2023.03.012

[11]

Xu H, Zhang Y, Liu Z, et al ETCM: an encyclopaedia of traditional Chinese medicine[J]. Nucleic Acids Res, 2019, 47: 976-982.

https://doi.org/10.1093/nar/gky987

[12]

Rappaport N, Twik M, Plaschkes I, et al MalaCards: an amalgamated human disease compendium with diverse clinical and genetic annotation and structured search[J]. Nucleic Acids Res, 2017, 45: 877-887.

https://doi.org/10.1093/nar/gkw1012

[13]

Safran M, Dalah I, Alexander J, et al GeneCards version 3: the human gene integrator[J]. Database, 2010, 2010: baq020.

[14]

Sayers EW, Beck J, Bolton EE, et al Database resources of the National Center for Biotechnology Information[J]. Nucleic Acids Res, 2021, 49: 10-17.

[15]

Barrett T, Wilhite SE, Ledoux P, et al NCBI GEO: archive for functional genomics data sets—update[J]. Nucleic Acids Res, 2013, 41: 991-995.

[16]

Zhang Y, Wang N, Du X, et al SoFDA: an integrated web platform from syndrome ontology to network-based evaluation of disease-syndrome-formula associations for precision medicine[J]. Sci Bull, 2022, 67: 1097-1101.

https://doi.org/10.1016/j.scib.2022.03.013

[17]

Wu Y, Zhang F, Yang K, et al SymMap: an integrative database of traditional Chinese medicine enhanced by symptom mapping[J]. Nucleic Acids Res, 2019, 47: 1110-1117.

https://doi.org/10.1093/nar/gky1260

[18]

Liu Y, Xu J, Yu Z, et al Ontology characterization, enrichment analysis, and similarity calculation-based evaluation of disease-syndrome-formula associations by applying SoFDA[J]. iMeta, 2023, 2: e80.

https://doi.org/10.1002/imt2.80

[19]

Szklarczyk D, Franceschini A, Wyder S, et al STRING v10: protein-protein interaction networks, integrated over the tree of life[J]. Nucleic Acids Res, 2015, 43: 447-452.

https://doi.org/10.1093/nar/gku1003

[20]

Dennis G Jr, Sherman BT, Hosack DA, et al DAVID: database for annotation, visualization, and integrated discovery[J]. Genome Biol, 2003, 4: P3.

https://doi.org/10.1186/gb-2003-4-5-p3

[21]

Coderre TJ, Wall PD Ankle joint urate arthritis (AJUA) in rats: an alternative animal model of arthritis to that produced by Freund's adjuvant[J]. Pain, 1987, 28: 379-393.

https://doi.org/10.1016/0304-3959(87)90072-8

[22]

Xiong H, Qu LY, Xiang LL, et al A rat model of gouty arthritis combined with dampness-heat syndrome[J]. J Tradit Chin Orthop Traumatol (中医正骨), 2014, 26: 14-20.

[23]

Coderre TJ, Wall PD Ankle joint urate arthritis in rats provides a useful tool for the evaluation of analgesic and anti-arthritic agents[J]. Pharmacol Biochem Behav, 1988, 29: 461-466.

https://doi.org/10.1016/0091-3057(88)90004-4

[24]

Guo ST. Observe Protective Effects and Mechanism of Qufengzhitongsan on Gouty Nephropathy and Acute Gouty Arthritis Rat (驱风止痛散对痛风性肾病及急性痛风性关节炎的保护作用及机制研究) [D]. Nanning: Guangxi Medical University, 2019.

[25]

Ma TH. Effect of Ethanol Extract of Polygonum cuspidatum on Acute Gouty Arthritis in Mice through NLRP3/ASC/Caspase-1 Signaling Pathway (虎杖醇提物通过NLRP3/ASC/Caspase-1轴干预小鼠急性痛风性关节炎的作用研究) [D]. Hangzhou: Zhejiang Chinese Medical University, 2018.

[26]

Guo Q, Zhao L, Zhu Y, et al Optimization of culture medium for Sanghuangporus vaninii and a study on its therapeutic effects on gout[J]. Biomed Pharmacother, 2021, 135: 111194.

https://doi.org/10.1016/j.biopha.2020.111194

[27]

Lv Y, Wang ZQ, Hou SX Establishment and observation of a rat model of lumbar facet joint-derived low back pain induced by intraspinal persistent compressive injury[J]. Chin J Pain Med (中国疼痛医学杂志), 2022, 28: 817-825.

[28]

Wang SS, Jiang ZL, Kuang WJ, et al Analgesic effect of AW2 in mice[J]. Sichuan J Physiol Sci (四川生理科学杂志), 2011, 33: 60-62.

[29]

Yuan X, Chen XL Traditional Chinese medicine's understanding of acute gouty arthritis[J]. Hubei J Tradit Chin Med (湖北中医杂志), 2019, 41: 62-66.

[30]

Liu ZH, Yang YF, Shi Y Contradiction and relationship between the cure and the treatment of gout Chinese medicine and Western medicine[J]. J Liaoning Univ Tradit Chin Med (辽宁中医药大学学报), 2020, 22: 64-67.

[31]

Liu XN, Gao ML Etilogy, pathogenesis and treatment of gouty arthritis with damp-heat accumulation[J]. Rheum Arthritis (风湿病与关节炎), 2022, 11: 43-45, 57.

[32]

Gan B, Li HN, Li S, et al Establishment of two rat models of gouty arthritis with damp-heat syndrome based on lipid metabolism and proinflammatory factors[J]. Chin J Comp Med (中国比较医学杂志), 2023, 33: 26-33.

[33]

Zhang P, Dou DB, Yu XP, et al Visual evaluation on curative effects of Chinese medicine on Yin-deficiency and dampness-heat syndrome in dampness-retention disease based on symptom value matrix[J]. J Tradit Chin Med (中医杂志), 2015, 56: 658-661.

[34]

Zou X. Clinical Study on the Treatment of Gouty Arthritis with Dampness and Heat Accumulation by Reducing Acid and Reducing Lipids (降酸降脂膏治疗湿热蕴结型痛风性关节炎的临床研究)[D]. Taiyuan: Shanxi University of Chinese Medicine, 2021.

[35]

Zhong YL, Xie GQ Exploration of the correlation between dampness heat syndrome and inflammatory factors in primary nephrotic syndrom[J]. J Liaoning Univ Tradit Chin Med (辽宁中医药大学学报), 2008, 10: 25-26.

[36]

Yin TT, Zhao CJ, Zhang YZ, et al Study on substance of damp-heat syndrome of rheumatic disease based on lipidomics[J]. China J Tradit Chin Med Pharm (中华中医药杂志), 2013, 28: 210-213.

[37]

Liu XY. Study on The Pathogenesis of Syndrome of Damp-heat of Non alcoholic Fatty Liver Disease Based on the Gut-liver Axi (基于"肠-肝"轴探讨非酒精性脂肪性肝病湿热证的发病机制)[D]. Chengdu: Chengdu University of Traditional Chinese Medicine, 2020.

[38]

Cao S, Liu N, Zhang M The relationship between gout and dyslipidemia in male patients[J]. Labeled Immunoassays Clin Med (标记免疫分析与临床), 2021, 28: 1646-1648, 1680.

[39]

Li HF, Lin PZ To approach mouse models of damp-heat syndrome of seasonal febrile disease infected by MHV-A59[J]. Liaoning J Tradit Chin Med (辽宁中医杂志), 2011, 38: 1318-1321.

[40]

Liu J, Xiao Q, Xiao J, et al Wnt/β-catenin signalling: function, biological mechanisms, and therapeutic opportunities[J]. Signal Transduct Target Ther, 2022, 7: 3.

https://doi.org/10.1038/s41392-021-00762-6

[41]

Schunk SJ, Floege J, Fliser D, et al Wnt-β-catenin signalling-a versatile player in kidney injury and repair[J]. Nat Rev Nephrol, 2021, 17: 172-184.

https://doi.org/10.1038/s41581-020-00343-w

[42]

Huang P, Yan R, Zhang X, et al Activating Wnt/β-catenin signaling pathway for disease therapy: challenges and opportunities[J]. Pharmacol Ther, 2019, 196: 79-90.

https://doi.org/10.1016/j.pharmthera.2018.11.008

[43]

Ma B, Hottiger MO Crosstalk between Wnt/β-catenin and NF-κB signaling pathway during inflammation[J]. Front Immunol, 2016, 7: 378.

[44]

Vallée A, Vallée JN, Lecarpentier Y Parkinson's disease: potential actions of lithium by targeting the Wnt/β-catenin pathway, oxidative stress, inflammation and glutamatergic pathway[J]. Cells, 2021, 10: 230.

https://doi.org/10.3390/cells10020230

[45]

Deng HH, Lin XL, Liu YH, et al Chitosan-stabilized platinum nanoparticles as effective oxidase mimics for colorimetric detection of acid phosphatase[J]. Nanoscale, 2017, 9: 10292-10300.

https://doi.org/10.1039/C7NR03399K

[46]

Liu J, Liao Y, Ma K, et al PI3KIs required for the physical interaction and functional inhibition of NF-κB by β-catenin in colorectal cancer cells[J]. Biochem Biophys Res Commun, 2013, 434: 760-766.

https://doi.org/10.1016/j.bbrc.2013.03.135

[47]

Martin M, Rehani K, Jope RS, et al Toll-like receptor-mediated cytokine production is differentially regulated by glycogen synthase kinase 3[J]. Nat Immunol, 2005, 6: 777-784.

[48]

Fliniaux I, Mikkola ML, Lefebvre S, et al Identification of Dkk4 as a target of Eda-A1/Edar pathway reveals an unexpected role of ectodysplasin as inhibitor of Wnt signalling in ectodermal placodes[J]. Dev Biol, 2008, 320: 60-71.

https://doi.org/10.1016/j.ydbio.2008.04.023

[49]

Yun K, Choi YD, Nam JH, et al NF-kappaB regulates Lef1 gene expression in chondrocytes[J]. Biochem Biophys Res Commun, 2007, 357: 589-595.

https://doi.org/10.1016/j.bbrc.2007.03.170

[50]

Prestwich TC, Macdougald OA Wnt/beta-catenin signaling in adipogenesis and metabolism[J]. Curr Opin Cell Biol, 2007, 19: 612-617.

https://doi.org/10.1016/j.ceb.2007.09.014

[51]

Kajimura S, Seale P, Spiegelman BM Transcriptional control of brown fat development[J]. Cell Metab, 2010, 11: 257-262.

https://doi.org/10.1016/j.cmet.2010.03.005

[52]

Morigny P, Boucher J, Arner P, et al Lipid and glucose metabolism in white adipocytes: pathways, dysfunction and therapeutics[J]. Nat Rev Endocrinol, 2021, 17: 276-295.

https://doi.org/10.1038/s41574-021-00471-8

[53]

Gu YQ. The Study on the Mechanism that Betaine Regulates ROS through Wnt/β-catenin Pathway in the ZF4 Cells of Zebrafish (甜菜碱通过Wnt/β-catenin通路调控斑马鱼ZF4细胞ROS的机制研究)[D]. Zibo: Shandong University of Technology, 2021.

[54]

Yang Z, Liu ZQ Role of PPARγ in adipocyte differentiation and glucose and lipid metabolism[J]. J Clin Pathol Res (临床与病理杂志), 2008, 28: 14-18.

2023年第58卷第8期

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doi: 10.16438/j.0513-4870.2023-0540

接收时间：2023-04-28
首发时间：2025-11-21
出版时间：2023-08-12

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收稿日期：2023-04-28
修回日期：2023-06-05

基金

国家自然科学基金青年科学基金项目(82204711)

中央本级重大增减支项目(2060302)

青年岐黄学者培养项目(国中医药人教函[2022]256号)

作者信息

中国中医科学院中药研究所, 北京 100700

通讯作者:

*张彦琼, E-mail: yqzhang@icmm.ac.cn;

许海玉, Tel: 86-10-64014411, E-mail: hyxu@icmm.ac.cn

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2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

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