原小檗碱类生物碱作用差异的机理研究
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摘要
原小檗碱类生物碱是一种异喹啉类生物碱，广泛存在于多种属植物体内，其药理作用广泛，其中抗菌、镇痛和降糖作用是临床常用的药理作用。我们通过实验观察原小檗碱型生物碱小檗碱、黄连碱、巴马汀、药根碱、延胡索乙素及罗通定的抑菌作用、镇痛作用和抗糖尿病作用的差异，并分别从理论上初步分析了其作用机理。
1．抑菌作用
采用平皿扩散法，研究了原小檗碱类生物碱对G~+菌(金黄色葡萄球菌、表皮葡萄球菌)、G~-菌(痢疾杆菌、肺炎克雷伯杆菌)标准菌株的抑菌作用。对于G~+菌金黄色葡萄球菌和表皮葡萄球菌，小檗碱的抑菌作用是巴马汀的3倍，是药根碱的10倍左右；对于痢疾杆菌的抑菌作用，小檗碱是黄连碱1.4倍，是巴马汀的10倍；对肺炎克雷伯菌仅小檗碱和黄连碱有抑菌作用，且小檗碱的抑菌作用是黄连碱的1.9倍。
分析结构表明，原小檗碱的季铵结构是抗菌活性所必须的结构，C2C3位的亚甲二氧基能增强抗菌活性。原小檗碱立体结构与四环素类和喹诺酮类结构叠合较好，提示作用机理有相似之处，可能通过抑制细菌核蛋白体30S亚基或DNA回旋酶A产生抑菌作用。配体-受体对接研究结果表明，原小檗碱与DNA回旋酶B和sortase B不匹配。
2．镇痛作用
给药1小时后，小鼠腹腔注射0.6％醋酸引起炎性疼痛，立即记录15分钟小鼠扭体次数，并计算药物对扭体反应的抑制率。罗痛定、延胡索乙素、小檗碱、巴马汀及黄连碱对扭体反应的抑制率分别为98.4，92.7，40.8，26.7，11.0％。罗通定、延胡索乙素和小檗碱的作用有统计学差异。分别测定给药前和单次灌
天津医科大学硕士研究生学位论灰
胃给药后30，60，120分钟的热板致痛阂值。罗通定和延胡索乙素在50mg/kg能
明显延长小鼠舔足时间，但小果碱、巴马汀和黄连碱150mg/kg对小鼠舔足时
间仍没影响。
理论计算表明，原小聚碱四氢化后(延胡索乙素，罗通定)，B一C环的二面
角以及N一CS，N一C14的键长发生改变，由此引起立体结构发生明显改变，镇痛
作用显著提高。四氢原小璧碱的镇痛作用与其它化合物不同，可能主要通过中
枢起作用。而小聚碱等原小栗碱季胺盐与环氧酶对接有合理性，提示可能是通
过抑制环氧酶而产生镇痛作用。分子动力学模拟显示，原小聚碱化合物与环氧
酶2主要是通过范德华作用结合，而且2，3位及9，10位的取代基在与受体结合
时贡献不同，这可能与其对化学性疼痛的抑制率不同相关。
3.降糖作用
尾静脉快速注射四氧喀咙SOm叭g制造糖尿病小鼠模型，连续5天灌胃
给药，末次给药2小时后测空腹血糖。小璧碱、黄连碱Zoomg/kg对四氧嚓咙
小鼠有降糖作用，且二者的降糖作用与二甲双肌比较差别无显著性印>0 .05)，巴
马汀则无降糖作用。
原小聚碱化合物CZ，C3位的亚甲二氧基可能与其降糖作用有关。原小聚
碱化合物与胰岛素增敏剂受体PPAR一丫对接结果与实验结果有一定的可比性，
认为原小聚碱可能通过PPAR一Y受体发挥降糖作用。进一步的分子动力学模拟
显示，原小栗碱主要是通过范德华相互作用与PPAR一Y蛋白产生作用。
总之，原小璧碱类生物碱结构差异很小，而作用面很广，作用差异较大，
对构效关系的研究是一个很大的挑战。我们结合理论和实验研究，找到了一些
可能的作用规律，但要真正阐明作用规律，还需要大量更深入的理论和实验研
究。
Protoberberine alkaloids, a kind of isoquinolone alkaloids, exist in several genera of the plant families. They have multiple pharmacological activities, especially the antibacterial, analgesic and anti-diabetic activities are the main effects that widely applied in clinic. The difference of the antibacterial, analgesic and anti-diabetic activities among berberine, palmatine, coptisine, jatrorrhizine, corydalis B and rotundine was studied. The mechanism of their function was analyzed by theoretical studies on molecular levels.
1. Antibacterial activity
The antimicrobial activity of protoberberines agaist the stardard strains of Gram-positive bacterium(s. aureus and s. epidermidis ) and Gram-negative bacterium (S. flexneri, E.coli and K.pnermoniae ) were tested using disc method with azithromycin and gentamicin as reference in vitro. The results showed that corydalis B exhibited no inhibitory activity against G+ and G- bacterium. The activities of berberine agaist S. aureus and S. epidermidis were about 3 and 10 times greater than that of berberine and jatrorrhizine respectively. Berberine was found to be 1.4 and 10 times more active aganist S. flexneri than coptisine and palmatine respectively. For K .pnermoniae, only berbeine and coptisine have antibacterial activities and the activity of berberine is 1.9 times than that of coptisine.
Structures analysis revealed that quaternary nitrogen is essential to antibacterial activity and the methylenedioxy group at the C2 and C3 is required for enhanced activity. The 3D structures of berberine , tetracyclines and quinolones have been compared and overlapped well ,which indicated probably their antibacterial mechanism is identical . The mechanism of antimicrobial of protoberberines maybe relate to the inhibition of 3 OS ribosome or DNA gyraseA . Molecular docking revealed that protoberberine alkaloids were not appropriate to DNA gyrase B or sortase B.
2. Analgesic activity
After 1 hour the drugs were administrated, the mice were intraperitoneally injected with 0.2ml of 0.6% acetic acid solution. The numbers of writhing reflex were counted for 15 minutes after the acetic acid injection. Rotundine , corydalis B and berberine could inhibit writhing movements of mice and the inhibitory rate of rotundine ,corydalis B, berberine ,palmatine and coptisine is 98.4, 92.7, 40.8, 26.7, 11.0% respectively. Licking paw time was measured before administration and 30,60,120min after single orally administrated. At dose of 50mg/kg ,rotundine and corydalis B had effects on mice licking paw time, but berberine, palmatine and coptisine have no effect at dose of 150mg/kg.
Rotundine and corydalis B of tetrahydroprotoberberines showed strong analgesic activity induced by chemical and heat stimulation. Theoretical calculation demonstrated that the dihedral angle of ring B and C and the bond lengths of N-C8, N-C14 strongly changed after reduction of the ring C, which lead to the three-dimensional structure changed strongly, thus the analgesic activity increased . The analgesic mechanism of tetrahydroprotoberberines was different to the other compounds, probably it is concerned to the central mechanism. The docking of other prtoberbennes with cyclo-oxygenase is rational ,which suggested the other alkoloids
of the proberberines maybe act as the cyclo-oxygenase inhibitors. Molecular dynamics simulations had been performed in Hyperchem 7.0, the results showed the protoberines mainly combined to Cox2 by Van der Waals , and the effects of substituent groups of 2,3 and 9,10 have different effects during ligand-receptor docking, which maybe relat to the analgesic activity induced by acetic acid.
3. Antidiabetic activity
Diabetic mice models were made after iv injected alloxan at dose of 80mg/kg. After orally administrated for 5 days , the blood samples were collected 2h after administration for determining fasting blood glucose with glucose oxidase method. Berberine and coptisine have anti-diabetic activities at dose of 200mg/kg. There was no significant difference of lowering glucose among b
1．抑菌作用
采用平皿扩散法，研究了原小檗碱类生物碱对G~+菌(金黄色葡萄球菌、表皮葡萄球菌)、G~-菌(痢疾杆菌、肺炎克雷伯杆菌)标准菌株的抑菌作用。对于G~+菌金黄色葡萄球菌和表皮葡萄球菌，小檗碱的抑菌作用是巴马汀的3倍，是药根碱的10倍左右；对于痢疾杆菌的抑菌作用，小檗碱是黄连碱1.4倍，是巴马汀的10倍；对肺炎克雷伯菌仅小檗碱和黄连碱有抑菌作用，且小檗碱的抑菌作用是黄连碱的1.9倍。
分析结构表明，原小檗碱的季铵结构是抗菌活性所必须的结构，C2C3位的亚甲二氧基能增强抗菌活性。原小檗碱立体结构与四环素类和喹诺酮类结构叠合较好，提示作用机理有相似之处，可能通过抑制细菌核蛋白体30S亚基或DNA回旋酶A产生抑菌作用。配体-受体对接研究结果表明，原小檗碱与DNA回旋酶B和sortase B不匹配。
2．镇痛作用
给药1小时后，小鼠腹腔注射0.6％醋酸引起炎性疼痛，立即记录15分钟小鼠扭体次数，并计算药物对扭体反应的抑制率。罗痛定、延胡索乙素、小檗碱、巴马汀及黄连碱对扭体反应的抑制率分别为98.4，92.7，40.8，26.7，11.0％。罗通定、延胡索乙素和小檗碱的作用有统计学差异。分别测定给药前和单次灌
天津医科大学硕士研究生学位论灰
胃给药后30，60，120分钟的热板致痛阂值。罗通定和延胡索乙素在50mg/kg能
明显延长小鼠舔足时间，但小果碱、巴马汀和黄连碱150mg/kg对小鼠舔足时
间仍没影响。
理论计算表明，原小聚碱四氢化后(延胡索乙素，罗通定)，B一C环的二面
角以及N一CS，N一C14的键长发生改变，由此引起立体结构发生明显改变，镇痛
作用显著提高。四氢原小璧碱的镇痛作用与其它化合物不同，可能主要通过中
枢起作用。而小聚碱等原小栗碱季胺盐与环氧酶对接有合理性，提示可能是通
过抑制环氧酶而产生镇痛作用。分子动力学模拟显示，原小聚碱化合物与环氧
酶2主要是通过范德华作用结合，而且2，3位及9，10位的取代基在与受体结合
时贡献不同，这可能与其对化学性疼痛的抑制率不同相关。
3.降糖作用
尾静脉快速注射四氧喀咙SOm叭g制造糖尿病小鼠模型，连续5天灌胃
给药，末次给药2小时后测空腹血糖。小璧碱、黄连碱Zoomg/kg对四氧嚓咙
小鼠有降糖作用，且二者的降糖作用与二甲双肌比较差别无显著性印>0 .05)，巴
马汀则无降糖作用。
原小聚碱化合物CZ，C3位的亚甲二氧基可能与其降糖作用有关。原小聚
碱化合物与胰岛素增敏剂受体PPAR一丫对接结果与实验结果有一定的可比性，
认为原小聚碱可能通过PPAR一Y受体发挥降糖作用。进一步的分子动力学模拟
显示，原小栗碱主要是通过范德华相互作用与PPAR一Y蛋白产生作用。
总之，原小璧碱类生物碱结构差异很小，而作用面很广，作用差异较大，
对构效关系的研究是一个很大的挑战。我们结合理论和实验研究，找到了一些
可能的作用规律，但要真正阐明作用规律，还需要大量更深入的理论和实验研
究。
Protoberberine alkaloids, a kind of isoquinolone alkaloids, exist in several genera of the plant families. They have multiple pharmacological activities, especially the antibacterial, analgesic and anti-diabetic activities are the main effects that widely applied in clinic. The difference of the antibacterial, analgesic and anti-diabetic activities among berberine, palmatine, coptisine, jatrorrhizine, corydalis B and rotundine was studied. The mechanism of their function was analyzed by theoretical studies on molecular levels.
1. Antibacterial activity
The antimicrobial activity of protoberberines agaist the stardard strains of Gram-positive bacterium(s. aureus and s. epidermidis ) and Gram-negative bacterium (S. flexneri, E.coli and K.pnermoniae ) were tested using disc method with azithromycin and gentamicin as reference in vitro. The results showed that corydalis B exhibited no inhibitory activity against G+ and G- bacterium. The activities of berberine agaist S. aureus and S. epidermidis were about 3 and 10 times greater than that of berberine and jatrorrhizine respectively. Berberine was found to be 1.4 and 10 times more active aganist S. flexneri than coptisine and palmatine respectively. For K .pnermoniae, only berbeine and coptisine have antibacterial activities and the activity of berberine is 1.9 times than that of coptisine.
Structures analysis revealed that quaternary nitrogen is essential to antibacterial activity and the methylenedioxy group at the C2 and C3 is required for enhanced activity. The 3D structures of berberine , tetracyclines and quinolones have been compared and overlapped well ,which indicated probably their antibacterial mechanism is identical . The mechanism of antimicrobial of protoberberines maybe relate to the inhibition of 3 OS ribosome or DNA gyraseA . Molecular docking revealed that protoberberine alkaloids were not appropriate to DNA gyrase B or sortase B.
2. Analgesic activity
After 1 hour the drugs were administrated, the mice were intraperitoneally injected with 0.2ml of 0.6% acetic acid solution. The numbers of writhing reflex were counted for 15 minutes after the acetic acid injection. Rotundine , corydalis B and berberine could inhibit writhing movements of mice and the inhibitory rate of rotundine ,corydalis B, berberine ,palmatine and coptisine is 98.4, 92.7, 40.8, 26.7, 11.0% respectively. Licking paw time was measured before administration and 30,60,120min after single orally administrated. At dose of 50mg/kg ,rotundine and corydalis B had effects on mice licking paw time, but berberine, palmatine and coptisine have no effect at dose of 150mg/kg.
Rotundine and corydalis B of tetrahydroprotoberberines showed strong analgesic activity induced by chemical and heat stimulation. Theoretical calculation demonstrated that the dihedral angle of ring B and C and the bond lengths of N-C8, N-C14 strongly changed after reduction of the ring C, which lead to the three-dimensional structure changed strongly, thus the analgesic activity increased . The analgesic mechanism of tetrahydroprotoberberines was different to the other compounds, probably it is concerned to the central mechanism. The docking of other prtoberbennes with cyclo-oxygenase is rational ,which suggested the other alkoloids
of the proberberines maybe act as the cyclo-oxygenase inhibitors. Molecular dynamics simulations had been performed in Hyperchem 7.0, the results showed the protoberines mainly combined to Cox2 by Van der Waals , and the effects of substituent groups of 2,3 and 9,10 have different effects during ligand-receptor docking, which maybe relat to the analgesic activity induced by acetic acid.
3. Antidiabetic activity
Diabetic mice models were made after iv injected alloxan at dose of 80mg/kg. After orally administrated for 5 days , the blood samples were collected 2h after administration for determining fasting blood glucose with glucose oxidase method. Berberine and coptisine have anti-diabetic activities at dose of 200mg/kg. There was no significant difference of lowering glucose among b
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44. Yoshinori Kobayashi, Yoshinori Yamashita, Noboru Fufii, et al. Inhibitors of DNA topoisomerase Ⅰ and Ⅱ isolated from the Coptis Rhizomes. Planta Med 1995,61:414-418
45. Krishnan P, Bastow KF. The 9-position in berberine analogs is an important determinant of DNA topoisomerase Ⅱ inhibition. AntiCancer Drug Des 2000, 15(4):255-264
46．赵长春，于俊生，章杰兵．药物与DNA的相互作用研究1小檗碱作为DNA的发光开关及Ph值的影响．无机化学学报，1997，13(3)：325-329
47. Soo-Hwan Kim, Dong-Sun Shin, Mi-Na Oh, et al. Inhibition of the bacterial surface protein anchoring transpeptidase sortase by isoquinoline alkaloids. Biosci Biotechnol Biochem, 2004, 68(2):421-424
48．金国章，郑秀风，胥彬．延胡索的药理研究Ⅶ．延胡索乙素旋光异构体的中枢作用．生理学报，1964,27(1)：47-58
49．金国章，周启霆，陈丽娟，俞蕾平等．四氢原小檗碱同类物(THPB)对DA受
体的新药理作用。中国科学基金，2000，(5)：300-304
50. Fukuda K, Hibiya Y, Muton M, et al. Inhibition by berberine of cyclooxygenase -2 transcriptional activity in human colon cancer cells. Journal of Ethnopharmacology 1999,66(2):227-233.
51．周丽斌、扬颖、唐金凤等．小檗碱对脂肪细胞糖代谢的影响．上海第二医科大学学报，2002，22(5)：412-414
52．陈其明，谢明智．小檗碱对正常小鼠血糖调节的影响．药学学报 1987，22(3)：161-165
53．殷峻，胡仁明，唐金风等．小檗碱的体外降螗作用．上海第二医科大学学报2001．21(5)：425-428。
54. Debanath D, Kumar GS, Nandi R et al. Interaction of berberine chloride with deoxyribonucleic acids: evidence for base and sequence specificity. Indian J Biophys, 1989,26(4):201-208,
55. Saran A, Srivastava S, Coutinho E ,et al. 1H NMR investigation of the interaction of berberine and sanguinarine with DNA. Indian J Biochem Biophys, 1995, 32(2): 74-77.
56. Debnath D, Suresh Kumar G, Maiti M. Circular dichroism studies of the structure of DNA complex with berberine. J Biomol Struct Dyn, 1991,9:61-79
57. Nandi R, Debnath D, Maiti M. Interactions of berberine with poly(A) and Trna. Biochim biophys acta. 1990,1049(3):339-342
58. Gong GQ, Zong ZX, Song YM. Spectrofluorometric determination of DNA and RNA with berberine. Spectrochim Acta A Mol Biomol Spectrosc, 1999, 55A (9): 1903-1907
59. Pilch DS, Yu c, Makhey D ,et al,. Minor groove-directed and intercalative ligand-DNA interactions in the poisoning of human DNA topoisomerase Ⅰ by protoberberine analogs. Biochemistry, 1997,36(41): 12542-12553
60. Mazmanian SK, Liu G, Ton-That H, et al. Staphylococcus aureus sortase mutants defective in the display of surface proteins and in the pathogenesis of animal infections. Proc Natl Acad Sci USA, 2000, 97(10): 5510-5515
61. Ilangovan U, Ton-That H, Iwahara J, et al.Clubb. Structure of sortase, the
tranpeptidase that anchors proteins to the cell wall of Staphylococcus aureus, PANS, 2001,98(11):6056-6061.
62. Ton-That H, Mazmanian SK, Alksne L, et al. Anchoring of surface proteins to the cell wall of Staphylococcus aureus. Cysteine 184 and histidine 120 of sortase form a thiolate-imidazolium ion pair for catalysis. J Biol Chem,2002,277(9):7447-7452.
63. Huang X, Aulabaugh A,Ding W, et al. Kinetic mechanism of Staphylococcus aureus sortase SrtA. Biochemisitry, 2003, 42 (38): 11307-11315.
64. Zong Y, Mazmanian SK, Schneewind O, et al. The structure of sortase B, a cysteine transpeptidase that tethers surface protein to the Staphylococcus aureus cell wall. Structure (Camb): 2004,12 (1): 105-112
65. Bierne H, Garandeau C, Pucciarelli MG, et al. Sortase B,a new class of sortase in Listeria monocytogenes. J Bacteriol. 2004, 186 (7): 1972-1982.
66. Mazmanian SK,Ton-That H,Su k.et al. An iron-regulated sortase anchors a class of surface protein during Staphylococcus aureus pathogenesis. PNAS,99(4):2293-2298
67. Kuo CL, Chi CW, Liu TY. The anti-inflammatory potential of berberine in vitro and in vivo. Cancer Lett. 2004,203 (2): 127-137
68．台卫平，罗和生，黄连素抑制结肠癌细胞还氧合酶-2的作用．中华内科杂志，2003，42(8)：558-560
69. Hu JY, Jin GZ. Effect of tetrahydropalmatine analogs on Fos expression induced by formalin-pain. Acta Pharmacol Sin, 1999, 20(3): 193-200
70. Hu JY, Jin GZ. Superspinal D2 receptor involved in antinociception induced by 1-tetrahydropalmatine. Acta Pharmacol Sin, 1999, 20:715-719
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73．高从荣，张家庆，黄庆玲．黄连素增加胰岛素抵抗大鼠模型胰岛素敏感性的实验研究．中西医结合杂志1997，17(3)：162-164．
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43. Li TK, Bathory E, LaVoie EJ et al. Human topoisomerase Ⅰ poising by protoberberines: potential roles for both drug-DNA and drug-enzyme interaction. Biochemistry 2000,39(24):7107-7116
44. Yoshinori Kobayashi, Yoshinori Yamashita, Noboru Fufii, et al. Inhibitors of DNA topoisomerase Ⅰ and Ⅱ isolated from the Coptis Rhizomes. Planta Med 1995,61:414-418
45. Krishnan P, Bastow KF. The 9-position in berberine analogs is an important determinant of DNA topoisomerase Ⅱ inhibition. AntiCancer Drug Des 2000, 15(4):255-264
46．赵长春，于俊生，章杰兵．药物与DNA的相互作用研究1小檗碱作为DNA的发光开关及Ph值的影响．无机化学学报，1997，13(3)：325-329
47. Soo-Hwan Kim, Dong-Sun Shin, Mi-Na Oh, et al. Inhibition of the bacterial surface protein anchoring transpeptidase sortase by isoquinoline alkaloids. Biosci Biotechnol Biochem, 2004, 68(2):421-424
48．金国章，郑秀风，胥彬．延胡索的药理研究Ⅶ．延胡索乙素旋光异构体的中枢作用．生理学报，1964,27(1)：47-58
49．金国章，周启霆，陈丽娟，俞蕾平等．四氢原小檗碱同类物(THPB)对DA受
体的新药理作用。中国科学基金，2000，(5)：300-304
50. Fukuda K, Hibiya Y, Muton M, et al. Inhibition by berberine of cyclooxygenase -2 transcriptional activity in human colon cancer cells. Journal of Ethnopharmacology 1999,66(2):227-233.
51．周丽斌、扬颖、唐金凤等．小檗碱对脂肪细胞糖代谢的影响．上海第二医科大学学报，2002，22(5)：412-414
52．陈其明，谢明智．小檗碱对正常小鼠血糖调节的影响．药学学报 1987，22(3)：161-165
53．殷峻，胡仁明，唐金风等．小檗碱的体外降螗作用．上海第二医科大学学报2001．21(5)：425-428。
54. Debanath D, Kumar GS, Nandi R et al. Interaction of berberine chloride with deoxyribonucleic acids: evidence for base and sequence specificity. Indian J Biophys, 1989,26(4):201-208,
55. Saran A, Srivastava S, Coutinho E ,et al. 1H NMR investigation of the interaction of berberine and sanguinarine with DNA. Indian J Biochem Biophys, 1995, 32(2): 74-77.
56. Debnath D, Suresh Kumar G, Maiti M. Circular dichroism studies of the structure of DNA complex with berberine. J Biomol Struct Dyn, 1991,9:61-79
57. Nandi R, Debnath D, Maiti M. Interactions of berberine with poly(A) and Trna. Biochim biophys acta. 1990,1049(3):339-342
58. Gong GQ, Zong ZX, Song YM. Spectrofluorometric determination of DNA and RNA with berberine. Spectrochim Acta A Mol Biomol Spectrosc, 1999, 55A (9): 1903-1907
59. Pilch DS, Yu c, Makhey D ,et al,. Minor groove-directed and intercalative ligand-DNA interactions in the poisoning of human DNA topoisomerase Ⅰ by protoberberine analogs. Biochemistry, 1997,36(41): 12542-12553
60. Mazmanian SK, Liu G, Ton-That H, et al. Staphylococcus aureus sortase mutants defective in the display of surface proteins and in the pathogenesis of animal infections. Proc Natl Acad Sci USA, 2000, 97(10): 5510-5515
61. Ilangovan U, Ton-That H, Iwahara J, et al.Clubb. Structure of sortase, the
tranpeptidase that anchors proteins to the cell wall of Staphylococcus aureus, PANS, 2001,98(11):6056-6061.
62. Ton-That H, Mazmanian SK, Alksne L, et al. Anchoring of surface proteins to the cell wall of Staphylococcus aureus. Cysteine 184 and histidine 120 of sortase form a thiolate-imidazolium ion pair for catalysis. J Biol Chem,2002,277(9):7447-7452.
63. Huang X, Aulabaugh A,Ding W, et al. Kinetic mechanism of Staphylococcus aureus sortase SrtA. Biochemisitry, 2003, 42 (38): 11307-11315.
64. Zong Y, Mazmanian SK, Schneewind O, et al. The structure of sortase B, a cysteine transpeptidase that tethers surface protein to the Staphylococcus aureus cell wall. Structure (Camb): 2004,12 (1): 105-112
65. Bierne H, Garandeau C, Pucciarelli MG, et al. Sortase B,a new class of sortase in Listeria monocytogenes. J Bacteriol. 2004, 186 (7): 1972-1982.
66. Mazmanian SK,Ton-That H,Su k.et al. An iron-regulated sortase anchors a class of surface protein during Staphylococcus aureus pathogenesis. PNAS,99(4):2293-2298
67. Kuo CL, Chi CW, Liu TY. The anti-inflammatory potential of berberine in vitro and in vivo. Cancer Lett. 2004,203 (2): 127-137
68．台卫平，罗和生，黄连素抑制结肠癌细胞还氧合酶-2的作用．中华内科杂志，2003，42(8)：558-560
69. Hu JY, Jin GZ. Effect of tetrahydropalmatine analogs on Fos expression induced by formalin-pain. Acta Pharmacol Sin, 1999, 20(3): 193-200
70. Hu JY, Jin GZ. Superspinal D2 receptor involved in antinociception induced by 1-tetrahydropalmatine. Acta Pharmacol Sin, 1999, 20:715-719
71. Storlin LH, Oakes ND, Pan D, et al. Syndromes of insulin resistance in the rat. Inducement by diet and the amelioration with benfluorex. Diabetes,1993, 42: 457-460.
72. Kraegen Ew, Clark PW. Jenkins AB, et al. Development of muscle insulin resistance after liver insulin resistance in high fat-fed rat. Diabetes 1991, 40: 1397-1403.
73．高从荣，张家庆，黄庆玲．黄连素增加胰岛素抵抗大鼠模型胰岛素敏感性的实验研究．中西医结合杂志1997，17(3)：162-164．