三脚风炉水提物对痛风性关节炎大鼠NLRP3炎性体及炎症因子的影响
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摘要
[目的]研究畲药三脚风炉水提物(Primpinellae diversifoliae water extracts,PDW)对痛风性关节炎大鼠Nod样受体蛋白3(Nod-like receptor pyrindomain containing 3,NLRP3)炎性体、炎症因子白细胞介素-1β(interleukin-1β,IL~(-1)β)及肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的影响,探讨PDW抗痛风性关节炎的作用机制。[方法]24只雄性SD大鼠随机分成正常对照组、模型组、美洛昔康组和PDW组。大鼠右踝关节皮下注射尿酸钠晶体制作急性痛风性关节炎模型,观察致炎前后踝关节肿胀度变化;Real-time PCR检测大鼠右踝关节组织中NLRP3、凋亡相关斑点样蛋白(apoptosis-associated speck-like protein,ASC)和半胱氨酸天冬氨酸蛋白酶-l(cysteinyl aspartate specific proteinase-1,caspase-1)的m RNA表达;ELISA法测定大鼠血清中炎症因子IL~(-1)β和TNF-α的含量。[结果]与模型组比较,PDW给药后5、24、48h能明显抑制痛风鼠踝关节肿胀,差异有统计学意义(P<0.001);PDW给药后,大鼠踝关节局部组织的NLRP3、ASC、caspase-1的mRNA表达水平显著下降,差异有统计学意义(P<0.001,P<0.01,P<0.01);大鼠血清中IL~(-1)β和TNF-α的含量显著下调,差异有统计学意义(P<0.001,P<0.01)。[结论]PDW具有一定的抗痛风性关节炎的作用,可能通过调节NLRP3炎性体表达,抑制炎症因子IL~(-1)β和TNF-α的分泌而发挥作用。
[Objective] To study anti-gout mechanism of of Primpinellae diversifoliae water extracts(PDW) by investigating its effects on the expression of Nod-like receptor pyrin domain containing 3(NLRP3) inflammasome and inflammatory cytokines levels of interleukin-1 β(IL~(-1)β) and tumor necrosis factor-α(TNF-α) in monosodium urate(MSU)-induced gouty arthritis rats. [Methods] Twenty four male SD rats were randomly divided into normal control group, model group, meloxicam group and PDW group. Gouty arthritis rats were induced by intradermal injection of MSU crystal suspension into the right ankle joint of rats. The anti-inflammatory activities were evaluated by paw edema. The mRNA expressions of NLRP3, apoptosis-associated speck-like protein(ASC) and cysteinyl aspartate specific proteinase-1(caspase-1) from the right ankle of rats were determined by Real-time PCR method; the levels of IL~(-1)β and TNF-α in serum of rats were detected by ELISA methods. [Results] Compared with model control, PDW inhibited the ankle swelling at 5 h,24 h and 48 h(P <0.001), downregulated the gene expression of NLRP3, ASC and caspase-1 in tissue of ankle joint( P <0.001, P <0.01, P <0.01), and significantly downregulated the serum level of IL~(-1)β and TNF-α(P<0.001, P<0.01). [Conclusion] PDW has the effects on gouty arthritis by inhibiting the paw edema which regulate NLRP3 inflammasome and reduce the levels of IL~(-1)β and TNF-α.
[Objective] To study anti-gout mechanism of of Primpinellae diversifoliae water extracts(PDW) by investigating its effects on the expression of Nod-like receptor pyrin domain containing 3(NLRP3) inflammasome and inflammatory cytokines levels of interleukin-1 β(IL~(-1)β) and tumor necrosis factor-α(TNF-α) in monosodium urate(MSU)-induced gouty arthritis rats. [Methods] Twenty four male SD rats were randomly divided into normal control group, model group, meloxicam group and PDW group. Gouty arthritis rats were induced by intradermal injection of MSU crystal suspension into the right ankle joint of rats. The anti-inflammatory activities were evaluated by paw edema. The mRNA expressions of NLRP3, apoptosis-associated speck-like protein(ASC) and cysteinyl aspartate specific proteinase-1(caspase-1) from the right ankle of rats were determined by Real-time PCR method; the levels of IL~(-1)β and TNF-α in serum of rats were detected by ELISA methods. [Results] Compared with model control, PDW inhibited the ankle swelling at 5 h,24 h and 48 h(P <0.001), downregulated the gene expression of NLRP3, ASC and caspase-1 in tissue of ankle joint( P <0.001, P <0.01, P <0.01), and significantly downregulated the serum level of IL~(-1)β and TNF-α(P<0.001, P<0.01). [Conclusion] PDW has the effects on gouty arthritis by inhibiting the paw edema which regulate NLRP3 inflammasome and reduce the levels of IL~(-1)β and TNF-α.
引文
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[4]浙江省食品药品监督管理局.浙江省中药炮制规范[M].杭州:浙江科学技术出版社,2005:557.Food and Drug Administration of Zhejiang Province. Tra-ditional Chinese medicine processing standards of Zhe-jiang Province[M].Hangzhou:Zhejiang Science and Tech-nology Press, 2005:557.
[5] Lv H, Zhang X, Chen X, et al. Phytochemical composi-tions and antioxidant and anti-inflammatory activities ofcrude extracts from Ficus pandurata H.(Moraceae)[J]. EvidBased Complement Alternat Med, 2013, 2013:215036.
[6]陈学智,温成平,李海昌,等.三角风炉抗炎镇痛及对Raw264.7细胞的体外抗炎活性[J].中华中医药学刊,2015,33(11):2595-2597.CHEN Xuezhi, WEN Chengping, LI Haichang, et al. An-ti-inflammatory and analgesic effects of Primpinellae di-versifoliae DC. and anti-inflammatory activities in LPS-stimulated RAW 264.7 cells in vitro[J]. Chinese Archivesof Traditional Chinese Medicine, 2015, 33(11):2595-2597.
[7]张昱.痛风[M].北京:科学技术文献出版社,2006:5-6.ZHANG Yu. Gout[M].Beijing:Science and Technology Lit-erature Press, 2006:5-6.
[8] Castelblanco M, Lugrin J, Ehirchiou D. Hydrogen sulfideinhibits NLRP3 inflammasome activation and reduces cy-tokine production both in vitro and in a mouse model ofinflammation[J]. J Biol Chem,2018,293(7):2546-2557.
[9] Yang G, Yeon S H, Lee H E, et al. Suppression of NL-RP3 inflammasome by oral treatment with sulforaphanealleviates acute gouty inflammation[J].Rheumatology(Ox-ford),2018, 57(4):727-736.
[2]雷后兴,李水福.中国畲族医药学[M].北京:中国中医药出版社,2007:10.LEI Houxing, LI Shuifu. Chinese Medicine of SHE Eth-nic Minority[M]. Beijing:China Press of Traditional Chi-nese Medicine, 2007:10.
[3]李水福.浅谈畲药三脚风炉[J].中国民族医药杂志,2009,15(9):47-48.LI Shuifu. Simple discussion of Primpinellae diversifoliaeDC. in SHE ethnic minority medicine[J]. Journal ofMedicine&Pharmacy of Chinese Minoritie, 2009, 15(9):47-48.
[4]浙江省食品药品监督管理局.浙江省中药炮制规范[M].杭州:浙江科学技术出版社,2005:557.Food and Drug Administration of Zhejiang Province. Tra-ditional Chinese medicine processing standards of Zhe-jiang Province[M].Hangzhou:Zhejiang Science and Tech-nology Press, 2005:557.
[5] Lv H, Zhang X, Chen X, et al. Phytochemical composi-tions and antioxidant and anti-inflammatory activities ofcrude extracts from Ficus pandurata H.(Moraceae)[J]. EvidBased Complement Alternat Med, 2013, 2013:215036.
[6]陈学智,温成平,李海昌,等.三角风炉抗炎镇痛及对Raw264.7细胞的体外抗炎活性[J].中华中医药学刊,2015,33(11):2595-2597.CHEN Xuezhi, WEN Chengping, LI Haichang, et al. An-ti-inflammatory and analgesic effects of Primpinellae di-versifoliae DC. and anti-inflammatory activities in LPS-stimulated RAW 264.7 cells in vitro[J]. Chinese Archivesof Traditional Chinese Medicine, 2015, 33(11):2595-2597.
[7]张昱.痛风[M].北京:科学技术文献出版社,2006:5-6.ZHANG Yu. Gout[M].Beijing:Science and Technology Lit-erature Press, 2006:5-6.
[8] Castelblanco M, Lugrin J, Ehirchiou D. Hydrogen sulfideinhibits NLRP3 inflammasome activation and reduces cy-tokine production both in vitro and in a mouse model ofinflammation[J]. J Biol Chem,2018,293(7):2546-2557.
[9] Yang G, Yeon S H, Lee H E, et al. Suppression of NL-RP3 inflammasome by oral treatment with sulforaphanealleviates acute gouty inflammation[J].Rheumatology(Ox-ford),2018, 57(4):727-736.