Establishment of Mice Nephritis Models and Observation of the Effects of Dihydroartemisinin on the Release of Inflammatory Cytokines_West China Medical Journal

Authors：

WU Ping ¹ , LI Qijie ² , XIA Zengliang ¹ , ZHANG Faqiang ¹ ,  XIA Qingjie ¹

1. State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P. R. China; 2. Basic Medical College, Chengdu University of Traditional Chinese Medicine, Chengdu, Sichuan 610075, P. R. China;

Corresponding author：

XIA Qingjie, Email: wuping.2008@163.com

Keywords：

肾炎模型; 双氢青蒿素; 脂多糖; 肿瘤坏死因子-α; 白细胞介素-6; 小鼠

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

【摘要】　目的　制备小鼠肾炎模型并观察双氢青蒿素（dihydroartemisinine，DHA）对模型小鼠细胞因子肿瘤坏死因子-α（tunor necrosis factor，TNF-α）和白细胞介素-6（inter leukin-6，IL-6）的影响以及小鼠肾脏的病理变化。　方法　取雄性昆明种小鼠120只，随机分为正常对照组、脂多糖（lipopolysaccharides，LPS）组、LPS+肾匀浆组及DHA治疗组；分别于12、24、48 h取血，酶联免疫吸附试验检测血清中TNF-α和IL-6的含量，苏木精-伊红染色法观察小鼠肾脏的病理变化。　结果　造模48 h LPS+肾匀浆组小鼠肾小球出现炎性细胞浸润，而正常对照组未见异常；LPS组及 DHA治疗组仅有轻微的病理改变。LPS刺激使小鼠血清TNF-α和IL-6含量高于正常水平（P lt;0.01），但有随时间不断下降的趋势；LPS+肾匀浆组较正常对照组TNF-α和IL-6含量升高（P lt;0.01）；DHA可显著下调模型小鼠血清TNF-α的水平（P lt;0.01），但对IL-6的影响相对较小（P gt;0.05）。　结论　运用改良的造模方法LPS+肾匀浆建立肾炎模型效果良好；DHA可以调节模型小鼠炎症因子TNF-α和IL-6的释放，具有一定的改善模型小鼠肾炎症状的作用。
【Abstract】　Objective　 To establish mice nephritis models, detect the serum level changes of cytokines tumor necrosis factor α (TNF-α) and interleukin-6 (IL-6) in the model mice treated by Dihydroartemisinin (DHA), and observe the physiological changes of the mice kidneys.　Methods　 One hundred and twenty male Kunming mice were randomly divided into 4 groups: control group, lipopolysaccharides (LPS) group, LPS plus kidney homogenate group, and DHA treated group. The level of cytokines TNF-α and IL-6 in the serum were detected by enzyme-linked immunoabsordent assey at hour 12, 24, and 48, respectively. Pathological changes were observed by hematoxylin: eosin staining.　Results　 At the time hour 48 after the establishment of the model, inflammatory cell infiltration was observed in the glomerulus of the LPS plus kidney homogenate group, but no abnormality was found in the control group. There were only slight pathological changes in mice models of the LPS group and the DHA treated group. The serum level of TNF-α and IL-6 increased remarkably after the treatment of LPS (P lt;0.01), but declined as time went by. The level of TNF-α and IL-6 increased significantly in LPS plus kidney homogenate group compared with the control group (P lt;0.01). DHA could significantly decrease the TNF-α level in the serum (P lt;0.01), but had a low influence on IL-6 (P gt;0.05).　Conclusion　 The modified LPS plus kidney homogenate has a good result in model establishing. DHA can regulate the release of TNF-α and IL-6 in the model mice, and may have certain good effects on ameliorating the nephritis pathological changes.

Citation： WU Ping,LI Qijie,XIA Zengliang,ZHANG Faqiang,XIA Qingjie. Establishment of Mice Nephritis Models and Observation of the Effects of Dihydroartemisinin on the Release of Inflammatory Cytokines. West China Medical Journal, 2011, 26(7): 1028-1031. doi: Copy

1.	仲芳, 陈慧, 韩琳, 等. 姜黄素对内毒素所致肾脏炎症的抑制作用[J]. 中国中西医结合肾病杂志, 2009, 10(11): 948-951.
2.	Zahner G, Schaper M, Panzer U, et al. Prostaglandin EP2 and EP4 receptors modulate expression of the chemokine CCL2 (MCP-1) in response to LPS-induced renal glomerular inflammation[J]. Biochem J, 2009, 422(3): 563-570.
3.	Shui HA, Ka SM, Wu WM, et al. LPS-evoked IL-18 expression in mesangial cells plays a role in accelerating lupus nephritis[J]. Rheumatology, 2007, 46(8): 1277-1284.
4.	Meyer-Schwesinger C, Dehde S, von Ruffer C, et al. Rho kinase inhibition attenuates LPS-induced renal failure in mice in part by attenuation of NF-kappa B p65 signaling[J]. Am J Physiol Renal Physiol, 2009, 296(5): 1088-1099.
5.	苑同业, 梁弘钢, 梁冰, 等. 肾小球肾炎患儿治疗前后TNFα、sTNFR、IL-6 和IL-8水平变化[J]. 上海医学检验杂志, 2003, 18(5): 260.
6.	段翠蓉, 李志辉. 白细胞介素-6与肾小球疾病的关系研究进展[J]. 中国当代医药, 2010, 17(16): 14-15.
7.	刘云海, 杜光. 青蒿素等中药化学成分抗内毒素研究现状[J]. 医药导报, 2001, 20(8): 525-526.
8.	梁爱华, 薛宝云, 王金华, 等. 青蒿琥酯对内毒素诱导的炎性因子合成的抑制作用的研究[J]. 中国中西医结合急救杂志, 2001, 8(5): 262.
9.	王俊, 周红. 青蒿素对CpG DNA攻击小鼠保护作用的实验研究[J]. 中国临床药理学与治疗学, 2005, 10(3): 290-293.
10.	董妍君, 李卫东, 屠呦呦, 等. 双氢青蒿素对BXSB狼疮小鼠自身抗体产生、TNFα分泌及狼疮性肾炎病理改变的影响[J]. 中国中西医结合杂志, 2003, 23(9): 676-679.
11.	赵桂芝, 聂淑琴, 杨庆, 等. 清脂胶囊对内毒素诱导的炎症介质水平的影响及时效关系[J]. 中国实验方剂学杂志, 2006, 12(11): 39-42.
12.	Sabry A, Elbasyouni SR, Sheashaa HA, et al. Correlation between levels of TNF-alpha and IL-6 and hematological involvement in SLE Egyptian Patients with lupus nephritis[J].Int Urol Nephrol, 2006, 38(3-4): 731-737.
13.	周立文, 贝文政, 王建才. 双氢青蒿素对恶性疟疾的疗效及免疫调节作用观察[J]. 热带病病与寄生虫学, 2008, 6(3): 138-139.
14.	Chen H, Sun B, Pan S, et al. Dihydroartemisinin inhibits growth of pancreatic cancer cells in vitro and in vivo[J].Anticancer Drugs, 2009, 20(2): 131-140.
15.	Noori S, Hassan ZM, Taghikhani M, et al. Dihydroartemisinin can inhibit calmodulin, calmodulin-dependent phosphodiesterase activity and stimulate cellular immune responses[J].Int Immunopharmacol, 2010, 10(2): 213-217.
16.	张荣侠, 吕晶晶, 戴夕超, 等. 双氢青蒿素对辐射小鼠血液系统的保护作用[J]. 中国现代医药杂志, 2009, 11(11): 39-41.
17.	李斌, 张乐之, 王俊, 等. 双氢青蒿素对CpG ODN诱导小鼠RAW264.7细胞释放细胞因子的影响[J]. 四川生理科学杂志, 2005, 27(4): 149-152.
18.	梁爱华, 薛宝云, 李春英, 等. 青蒿琥酯对内毒素诱导的一氧化氮合成的抑制作用[J]. 国外医学•中医中药分册, 2001, 23(3): 156.
19.	Natanson C, Hoffman WD, Suffredini AF, et al. Selected treatment strategies for septic shock based on proposed mechanisms of pathogenesis[J]. Ann Intern Med, 1994, 120(9): 771-783.
20.	Noda T, Amano F. Differences in nitric oxide synthase activity in a macrophage-like cell line, RAW264.7 cells, treated with lipopolysaccharide (LPS) in the presence or absence of interferon-gamma (IFN-gamma): possible heterogeneity of iNOS activity[J]. J Biochem, 1997, 121(1): 38-46.
21.	霍汝亚寒. 急性肾炎患儿治疗前后血清hs-CRP、lL-2、lL-6和GM-CSF检测的临床意义[J]. 放射免疫学杂志, 2010, 23(2): 133-134.
22.	Aldieri E, Atragene D, Bergandi L, et al. Artemisinin inhibits inducible nitric oxide synthase and nuclear factor NF-κB activation[J]. FEBS Lett, 2003, 552(2-3): 141-144.

1. 　仲芳, 陈慧, 韩琳, 等. 姜黄素对内毒素所致肾脏炎症的抑制作用[J]. 中国中西医结合肾病杂志, 2009, 10(11): 948-951.
2. 　Zahner G, Schaper M, Panzer U, et al. Prostaglandin EP2 and EP4 receptors modulate expression of the chemokine CCL2 (MCP-1) in response to LPS-induced renal glomerular inflammation[J]. Biochem J, 2009, 422(3): 563-570.
3. 　Shui HA, Ka SM, Wu WM, et al. LPS-evoked IL-18 expression in mesangial cells plays a role in accelerating lupus nephritis[J]. Rheumatology, 2007, 46(8): 1277-1284.
4. 　Meyer-Schwesinger C, Dehde S, von Ruffer C, et al. Rho kinase inhibition attenuates LPS-induced renal failure in mice in part by attenuation of NF-kappa B p65 signaling[J]. Am J Physiol Renal Physiol, 2009, 296(5): 1088-1099.
5. 　苑同业, 梁弘钢, 梁冰, 等. 肾小球肾炎患儿治疗前后TNFα、sTNFR、IL-6 和IL-8水平变化[J]. 上海医学检验杂志, 2003, 18(5): 260.
6. 　段翠蓉, 李志辉. 白细胞介素-6与肾小球疾病的关系研究进展[J]. 中国当代医药, 2010, 17(16): 14-15.
7. 　刘云海, 杜光. 青蒿素等中药化学成分抗内毒素研究现状[J]. 医药导报, 2001, 20(8): 525-526.
8. 　梁爱华, 薛宝云, 王金华, 等. 青蒿琥酯对内毒素诱导的炎性因子合成的抑制作用的研究[J]. 中国中西医结合急救杂志, 2001, 8(5): 262.
9. 　王俊, 周红. 青蒿素对CpG DNA攻击小鼠保护作用的实验研究[J]. 中国临床药理学与治疗学, 2005, 10(3): 290-293.
10. 　董妍君, 李卫东, 屠呦呦, 等. 双氢青蒿素对BXSB狼疮小鼠自身抗体产生、TNFα分泌及狼疮性肾炎病理改变的影响[J]. 中国中西医结合杂志, 2003, 23(9): 676-679.
11. 　赵桂芝, 聂淑琴, 杨庆, 等. 清脂胶囊对内毒素诱导的炎症介质水平的影响及时效关系[J]. 中国实验方剂学杂志, 2006, 12(11): 39-42.
12. 　Sabry A, Elbasyouni SR, Sheashaa HA, et al. Correlation between levels of TNF-alpha and IL-6 and hematological involvement in SLE Egyptian Patients with lupus nephritis[J].Int Urol Nephrol, 2006, 38(3-4): 731-737.
13. 　周立文, 贝文政, 王建才. 双氢青蒿素对恶性疟疾的疗效及免疫调节作用观察[J]. 热带病病与寄生虫学, 2008, 6(3): 138-139.
14. 　Chen H, Sun B, Pan S, et al. Dihydroartemisinin inhibits growth of pancreatic cancer cells in vitro and in vivo[J].Anticancer Drugs, 2009, 20(2): 131-140.
15. 　Noori S, Hassan ZM, Taghikhani M, et al. Dihydroartemisinin can inhibit calmodulin, calmodulin-dependent phosphodiesterase activity and stimulate cellular immune responses[J].Int Immunopharmacol, 2010, 10(2): 213-217.
16. 　张荣侠, 吕晶晶, 戴夕超, 等. 双氢青蒿素对辐射小鼠血液系统的保护作用[J]. 中国现代医药杂志, 2009, 11(11): 39-41.
17. 　李斌, 张乐之, 王俊, 等. 双氢青蒿素对CpG ODN诱导小鼠RAW264.7细胞释放细胞因子的影响[J]. 四川生理科学杂志, 2005, 27(4): 149-152.
18. 　梁爱华, 薛宝云, 李春英, 等. 青蒿琥酯对内毒素诱导的一氧化氮合成的抑制作用[J]. 国外医学•中医中药分册, 2001, 23(3): 156.
19. 　Natanson C, Hoffman WD, Suffredini AF, et al. Selected treatment strategies for septic shock based on proposed mechanisms of pathogenesis[J]. Ann Intern Med, 1994, 120(9): 771-783.
20. 　Noda T, Amano F. Differences in nitric oxide synthase activity in a macrophage-like cell line, RAW264.7 cells, treated with lipopolysaccharide (LPS) in the presence or absence of interferon-gamma (IFN-gamma): possible heterogeneity of iNOS activity[J]. J Biochem, 1997, 121(1): 38-46.
21. 　霍汝亚寒. 急性肾炎患儿治疗前后血清hs-CRP、lL-2、lL-6和GM-CSF检测的临床意义[J]. 放射免疫学杂志, 2010, 23(2): 133-134.
22. 　Aldieri E, Atragene D, Bergandi L, et al. Artemisinin inhibits inducible nitric oxide synthase and nuclear factor NF-κB activation[J]. FEBS Lett, 2003, 552(2-3): 141-144.

West China Medical Journal

Establishment of Mice Nephritis Models and Observation of the Effects of Dihydroartemisinin on the Release of Inflammatory Cytokines

Abstract Full text Figures/Tables Video References Cited by

Format

Content