• 1 四川大學(xué)華西醫(yī)院生物治療國家重點實驗室（成都，610041）;2成都中醫(yī)藥大學(xué)基礎(chǔ)醫(yī)學(xué)院;

【摘要】　目的　 制備小鼠腎炎模型并觀察雙氫青蒿素（dihydroartemisinine，DHA）對模型小鼠細胞因子腫瘤壞死因子-α（tunor necrosis factor，TNF-α）和白細胞介素-6（inter leukin-6，IL-6）的影響以及小鼠腎臟的病理變化。　方法　 取雄性昆明種小鼠120只， 隨機分為正常對照組、脂多糖（lipopolysaccharides，LPS）組、LPS+腎勻漿組及DHA治療組；分別于12、24、48 h取血，酶聯(lián)免疫吸附試驗檢測血清中TNF-α和IL-6的含量，蘇木精-伊紅染色法觀察小鼠腎臟的病理變化。　結(jié)果　 造模48 h LPS+腎勻漿組小鼠腎小球出現(xiàn)炎性細胞浸潤，而正常對照組未見異常；LPS組及 DHA治療組僅有輕微的病理改變。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），但有隨時間不斷下降的趨勢；LPS+腎勻漿組較正常對照組TNF-α和IL-6含量升高（P lt;0.01）；DHA可顯著下調(diào)模型小鼠血清TNF-α的水平（P lt;0.01），但對IL-6的影響相對較?。≒ gt;0.05）。　結(jié)論　 運用改良的造模方法LPS+腎勻漿建立腎炎模型效果良好；DHA可以調(diào)節(jié)模型小鼠炎癥因子TNF-α和IL-6的釋放，具有一定的改善模型小鼠腎炎癥狀的作用。
【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.

引用本文： 吳蘋,李啟杰,夏増亮,張發(fā)強,夏慶杰. 小鼠腎炎模型的制備及雙氫青蒿素對其炎癥因子釋放的影響. 華西醫(yī)學(xué), 2011, 26(7): 1028-1031. doi: 復(fù)制

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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. 　苑同業(yè), 梁弘鋼, 梁冰, 等. 腎小球腎炎患兒治療前后TNFα、sTNFR、IL-6 和IL-8水平變化[J]. 上海醫(yī)學(xué)檢驗雜志, 2003, 18(5): 260.
6. 　段翠蓉, 李志輝. 白細胞介素-6與腎小球疾病的關(guān)系研究進展[J]. 中國當代醫(yī)藥, 2010, 17(16): 14-15.
7. 　劉云海, 杜光. 青蒿素等中藥化學(xué)成分抗內(nèi)毒素研究現(xiàn)狀[J]. 醫(yī)藥導(dǎo)報, 2001, 20(8): 525-526.
8. 　梁愛華, 薛寶云, 王金華, 等. 青蒿琥酯對內(nèi)毒素誘導(dǎo)的炎性因子合成的抑制作用的研究[J]. 中國中西醫(yī)結(jié)合急救雜志, 2001, 8(5): 262.
9. 　王俊, 周紅. 青蒿素對CpG DNA攻擊小鼠保護作用的實驗研究[J]. 中國臨床藥理學(xué)與治療學(xué), 2005, 10(3): 290-293.
10. 　董妍君, 李衛(wèi)東, 屠呦呦, 等. 雙氫青蒿素對BXSB狼瘡小鼠自身抗體產(chǎn)生、TNFα分泌及狼瘡性腎炎病理改變的影響[J]. 中國中西醫(yī)結(jié)合雜志, 2003, 23(9): 676-679.
11. 　趙桂芝, 聶淑琴, 楊慶, 等. 清脂膠囊對內(nèi)毒素誘導(dǎo)的炎癥介質(zhì)水平的影響及時效關(guān)系[J]. 中國實驗方劑學(xué)雜志, 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, 貝文政, 王建才. 雙氫青蒿素對惡性瘧疾的療效及免疫調(diào)節(jié)作用觀察[J]. 熱帶病病與寄生蟲學(xué), 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. 　張榮俠, 呂晶晶, 戴夕超, 等. 雙氫青蒿素對輻射小鼠血液系統(tǒng)的保護作用[J]. 中國現(xiàn)代醫(yī)藥雜志, 2009, 11(11): 39-41.
17. 　李斌, 張樂之, 王俊, 等. 雙氫青蒿素對CpG ODN誘導(dǎo)小鼠RAW264.7細胞釋放細胞因子的影響[J]. 四川生理科學(xué)雜志, 2005, 27(4): 149-152.
18. 　梁愛華, 薛寶云, 李春英, 等. 青蒿琥酯對內(nèi)毒素誘導(dǎo)的一氧化氮合成的抑制作用[J]. 國外醫(yī)學(xué)?中醫(yī)中藥分冊, 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. 　霍汝亞寒. 急性腎炎患兒治療前后血清hs-CRP、lL-2、lL-6和GM-CSF檢測的臨床意義[J]. 放射免疫學(xué)雜志, 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. 1. 　仲芳, 陳慧, 韓琳, 等. 姜黃素對內(nèi)毒素所致腎臟炎癥的抑制作用[J]. 中國中西醫(yī)結(jié)合腎病雜志, 2009, 10(11): 948-951.
  2. 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. 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. 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. 5. 　苑同業(yè), 梁弘鋼, 梁冰, 等. 腎小球腎炎患兒治療前后TNFα、sTNFR、IL-6 和IL-8水平變化[J]. 上海醫(yī)學(xué)檢驗雜志, 2003, 18(5): 260.
  6. 6. 　段翠蓉, 李志輝. 白細胞介素-6與腎小球疾病的關(guān)系研究進展[J]. 中國當代醫(yī)藥, 2010, 17(16): 14-15.
  7. 7. 　劉云海, 杜光. 青蒿素等中藥化學(xué)成分抗內(nèi)毒素研究現(xiàn)狀[J]. 醫(yī)藥導(dǎo)報, 2001, 20(8): 525-526.
  8. 8. 　梁愛華, 薛寶云, 王金華, 等. 青蒿琥酯對內(nèi)毒素誘導(dǎo)的炎性因子合成的抑制作用的研究[J]. 中國中西醫(yī)結(jié)合急救雜志, 2001, 8(5): 262.
  9. 9. 　王俊, 周紅. 青蒿素對CpG DNA攻擊小鼠保護作用的實驗研究[J]. 中國臨床藥理學(xué)與治療學(xué), 2005, 10(3): 290-293.
  10. 10. 　董妍君, 李衛(wèi)東, 屠呦呦, 等. 雙氫青蒿素對BXSB狼瘡小鼠自身抗體產(chǎn)生、TNFα分泌及狼瘡性腎炎病理改變的影響[J]. 中國中西醫(yī)結(jié)合雜志, 2003, 23(9): 676-679.
  11. 11. 　趙桂芝, 聶淑琴, 楊慶, 等. 清脂膠囊對內(nèi)毒素誘導(dǎo)的炎癥介質(zhì)水平的影響及時效關(guān)系[J]. 中國實驗方劑學(xué)雜志, 2006, 12(11): 39-42.
  12. 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. 13. 　周立文, 貝文政, 王建才. 雙氫青蒿素對惡性瘧疾的療效及免疫調(diào)節(jié)作用觀察[J]. 熱帶病病與寄生蟲學(xué), 2008, 6(3): 138-139.
  14. 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. 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. 16. 　張榮俠, 呂晶晶, 戴夕超, 等. 雙氫青蒿素對輻射小鼠血液系統(tǒng)的保護作用[J]. 中國現(xiàn)代醫(yī)藥雜志, 2009, 11(11): 39-41.
  17. 17. 　李斌, 張樂之, 王俊, 等. 雙氫青蒿素對CpG ODN誘導(dǎo)小鼠RAW264.7細胞釋放細胞因子的影響[J]. 四川生理科學(xué)雜志, 2005, 27(4): 149-152.
  18. 18. 　梁愛華, 薛寶云, 李春英, 等. 青蒿琥酯對內(nèi)毒素誘導(dǎo)的一氧化氮合成的抑制作用[J]. 國外醫(yī)學(xué)?中醫(yī)中藥分冊, 2001, 23(3): 156.
  19. 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. 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. 21. 　霍汝亞寒. 急性腎炎患兒治療前后血清hs-CRP、lL-2、lL-6和GM-CSF檢測的臨床意義[J]. 放射免疫學(xué)雜志, 2010, 23(2): 133-134.
  22. 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.