The effect of Kuijieling decoction on the protein expression of I B and IKKβ in colonic mucosa of ulcerative colitis model rats

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137 6.. 11 Grivennikov SI Karin M. Dangerous liaisons STAT3 and NF-κB 2006 14 4 240 ~ 242 7 Vallabhapurapu S Karin M. Regulation and function of NF-kappa B transcription factors in the immune system. Annu Rev Immunol 2009 27 collab oration and crosstalk in cancer. Cytok Growth 2010 21 1 11 ~ 19 12 Caamano J Hunter CA. NF-κB family of transcription factors central 5 693 ~ 733 regulators of innate and adaptive immune functions. Clin Microbiol Rev 8 Benitah SA Valeron PF Lacal JC. Rock and nuclear factor kappabdependent 2002 15 414 ~ 429 activation of cyclooxygenase-2 by Rho GTPases effects on tumor 13 Klement J F Rice N R Car BD. I B deficiency results in a sustained growth and therapeutic consequences. Mol Biol Cell 2003 14 7 3041 ~ NF-κB response and severe widespread dermatitis in mice. Mol Cell Biol 3054 1996 16 2341 ~ 2349 9 Li Z Zhang DK Yi WQ et al. NF-kappaB p65 antisense oligonucleotides 14. NF-κB. 2003 may serve as a novel molecular approach for the treatment of patients 48 18 1893 ~ 1911 with ulcerative colitis. Arch Med Res 2008 39 8 729 ~ 734 15 Gavrilyuk V Dello Russo C Heneka MT. Norepinephrine increase I 10. kappa B alpha expression in astrocytes. J Biol Chem 2002 277 33 29662. 2002 82 6 384 ~ ~ 29668 388 The effect of Kuijieling decoction on the protein expression of I B and IKKβ in colonic mucosa of ulcerative colitis model rats Tang Lihai Ji Yanyan Du Qun Li Yanwu Wu Yanli Chai Yuna Chen Xu Wang Rujun Pi-Wei Institute Guangzhou University of Chinese Medicine Guangzhou 510405 Objective To observe the effect of Kuijieling decoction KD on protein expression of I B and IKKβin colonic mucosa of rats with ulcerative colitis UC and to explore its possible mechanism. Methods Trinitrobenzenesulfonic acid TNBS method was used for the establishment of UC rat model. After treatment with different dosages of KD Western blot technique were used to detect protein expression of I B and IKKβin Colonic Mucosa. Results The relative expression of I B protein in model group was obviously lower than that in normal group P < 0. 05 the relative expression of I B protein in high dosage group of KD was significantly higher than that in model group P < 0. 01 the relative expression of I B protein in middle dosage group of KD was obviously higher than that in model group P < 0. 01 The relative expression of IKKβprotein in model group was significantly higher than that in normal group P < 0. 01 the relative expression of IKKβprotein in both high and middle dosage groups of KD were significantly lower than that in model group P < 0. 05. Conclusion The relative expression of I B protein is enhanced and the relative expression of IKKβprotein is Inhibited in colonic mucosa of UC model rats induced by TNBS after treatment with KD Its possible mechanism may inhibit the activation of NF-κB and reduce inflammatory reaction. Key words Kuijieling decoction ulcerative colitis I B IKKβ rat model * 1 1 1** 2 2 3 4 2 1 201203 2 201203 3 201203 4 201203 4 4 4 HE * 2009ZX09311-003 **

138 10g /kg 6g /kg centre 3 Thermo Shandon Finesse E + Thermo Fisher 1. 5 1 10% CCl 4 2ml /kg 3 4 10g /kg N- -L- 0. 15g /kg 1 4 DMN DMN 10μl /kg 1 3 4 1 2 /3 2 4 6g /kg 5mg /kg 1 4 1. 5. 1 1. 5. 2 ALT AST Carbon tetrachloride CCl 4 Dimeth- Alb ylnitrosamine DMN 1. 5. 3 Hyp " μg /g " 1. 5. 4 HE 60 60min I 10min 1 II 10min I 5min II 2min 95% 2min 85% 2min 70% 2min 6 5 ~ 6 1. 1 15min 6 3s 4s 95% I 1min 95% 2 II 1min I 1min II 1min I 1min 1 10 OlympusBX40 200 Im- Express6. 3 8 age-pro 20100395 N- 1. 5. 5 60 60min I -L- 20100311 10min II 10min I 5min II 2min 95% 2min 85% 2min 70% 2min 6 1. 2 C57BL /6 10 25 ± 5 ~ 6 2g Wistar 150 ~ 180g 37 25min SCXK 2007-0005 OlympusBX40 100 Image-pro Express6. 3 1. 3 CCl 4 20100308 F20100311 DMN KWM 6890 2 Alanine aminotransferase ALT 20101011 Aspartate aminotransferaseast 20101009 Albumin Alb 20101008 20110303 Hydroxyproline Hyp P < 0. 01 20101028 P < 0. 05 1 1. 4 Varifuge 20RS HERAEUS Synergy HT Bio-tek Thermo Shandon Citadel 2000 Thermo Shandon Histo- 2. 1 CCl 4 P < 0. 01 DMN P < 0. 01 P < 0. 01 P > 0. 05 2

139 1 CCl 4 Hyp x 珋 ± s n = 10 P < 0. 01 3 4 3 CCl 4 Hyp g % x 珋 ± s n = 10 26. 70 ± 0. 95** 3. 52 ± 0. 36** 3. 73 ± 0. 59* ALT AST Alb Hyp 23. 80 ± 0. 79 4. 46 ± 0. 43 5. 70 ± 1. 31 6 25. 40 ± 1. 35 4. 57 ± 0. 62 4. 26 ± 0. g/kg IU /L IU /L g/l μg/g 76** N- -L- 106. 14 ± 31. 72** 145. 11 ± 42. 34** 33. 57 ± 1. 94 214. 40 ± 31. 71** 0. 15 23. 60 ± 0. 84 4. 11 ± 0. 64 5. 17 ±. 013 191. 72 ± 44. 24 232. 55 ± 58. 11 31. 56 ± 2. 29 309. 40 ± 57. 04 * P < 0. 05 ** P < 0. 01 6 129. 79 ± 47. 98* 168. 72 ± 40. 24* 32. 51 ± 2. 12 250. 77 ± 34. 87* 2 DMN N- -L- 0. 15 169. 30 ± 51. 41 189. 82 ± 66. 70 33. 20 ± 2. 71 261. 62 ± 68. 94 x 珋 ± s n = 10 g % 1. 68 ± 0. 43** 327. 20 ± 19. 62** 25. 26 ± 1. 43 290. 50 ± 7. 84 27. 75 ± 1. 65 2. 66 ± 0. 34 6 300. 80 ± 10. 42 27. 38 ± 0. 89 2. 58 ± 0. 29 0. 005 277. 62 ± 24. 60 27. 49 ± 2. 08 2. 61 ± 0. 28 4 DMN Hyp x 珋 ± s n = 10 6 43. 76 ± 11. 05** 146. 00 ± 25. 14* 30. 72 ± 2. 06 174. 40 ± 13. 95** 2. 2 0. 15 105. 99 ± 18. 35 241. 55 ± 29. 38 30. 28 ± 3. 26 215. 23 ± 47. 73* ALT AST P < 0. 2. 4 01 Alb P > 0. 05 HE ALT AST P < 0. 05 P < 0. 01 34 2. 3 Hyp Hyp P < 0. 01 g/kg ALT IU /L AST IU /L Alb g/l Hyp μg/g 41. 35 ± 15. 89** 128. 70 ± 46. 53** 33. 17 ± 2. 68 192. 54 ± 55. 23** 109. 92 ± 36. 42 256. 13 ± 52. 73 31. 04 ± 2. 33 291. 49 ± 76. 62 2. 5 P < 0. 05 56 3 CCl 4 100

140 4 DMN 100 5 = 10 CCl 4 x 珋 ± s n 0 I II III IV Ridit 10 0. 05** 1 5 4 0. 64 6 3 5 2 0. 50 N- -L- 0. 15 1 6 3 0. 61 ** P < 0. 01 6 = 10 DMN x 珋 ± s n 0 I II III IV Ridit 10 0. 06** 1 6 3 0. 70 6 4 6 0 0. 48* DMN DMN 3 TGF-β1 -α 4 TGF-β1 CTGF PDGF-BB 5 DMN 0. 15 1 8 1 0. 63 6 A 7 TGF-β1 8 3 " " " " " " " " " " 1 Svetlana R Rui S Barbara J et al. Natural killer cells ameliorate liver fibrosis by killing activated stellate cells in NKG2D-dependent and tumor nec- " " " " rosis factor-related apoptosis-inducing ligand-dependent manners. Gastroenterology 2006 130 2 435 ~ 452 CCl 4 2 Ala-kokko L Pihlajaniemi T Myers J et al. Gene expression of type I DMN III and Ⅳ collagens in hepatic fibrosis induced by dimethylnitrosamine in the rat. Biochem J 1987 244 1 75 ~ 79 3.. 2008 42 5 92 ~ 94 ALT AST 4. TNF-α TGFβ-1. 2011 32 6 758 ~ 759 5.. 2007 15 2 99 ~ 101 6. CCl 4. 2008 19 7 1565 ~ 1567 7. HSC-T6. 2009 40 8 1270 ~ 1274 DMN 8. A TGF-β1. 2010 33 6 405 ~ 408

141 The effects of Fugan decoction on hepatic fibrosis models in mouse and rat Cui Hongyan 1 Gu Weiliang 1 Ma Yueming 1 Liu Ping 2 Liu Chenghai 2 Zhu Bangxian 3 Wang Zhengtao 4 Tao Yanyan 2 1 Department of Pharmacology Shanghai University of Traditional Chinese Medicine Shanghai 201203 2 Institute of Liver Disease Shanghai University of Traditional Chinese Medicine Shanghai 201203 3 Information Center for Life Science of Chinese Academy of Sciences Shanghai University of Traditional Chinese Medicine Shanghai 201203 4 Institute of Traditional Chinese Medicine Shanghai University of Traditional Chinese Medicine Shanghai 201203 Objective To evaluate the preventive and therapeutic effects of Fugan Decoction on liver fibrosis induced by carbon tetrachloride CCl 4 in mice and by dimethylnitrosamine DMN in rats. Methods Liver fibrosis in mice was induced by intraperitoneal injection of CCl 4. The rats were randomly divided into 6 groups normal model and Fugan Low Fugan middle Fugan High and N-Acetyl-L-Cysteine control group. All the mice were administrated drugs by garvage for 4 weeks from the the fist CCl 4. injection day. Liver fibrosis in rats was induced by intraperitoneal injection of dimethylnitrosamine DMN. The rats were randomly divided into 6 groups normal model and Fugan Low Fugan middle Fugan High and Perindopril control group. The rats were administered drugs by garvage for 4 weeks after the model was established. The ratios of liver weight /body weight and spleen weight /body weight were calculated. Serum ALT AST activity were determined by colorimetry Alb levels were assayed by bromocresol green method serum hepatic inflammation and collagen deposition were evaluated with HE staining and Sirius-red staining respectively the contents of hepatic hydroxyproline Hyp were measured by alkali digestion method. Results Abnormal liver function obvious hepatic inflammation excess collagen deposition were observed in model group both of mouse and rat compared with those in normal group. Compared to model group Fugan Decoction significantly down-regulated serum ALT and AST activity alleviated liver inflammation it also reduced the level of hepatic hydroxyproline and the pathological excess collagen deposition in liver. Conclusion Fugan decoction has good preventive and therapeutic effects in experimental liver fibrosis. Key words Fugan carbontetrachloride dimethylnitrosamine liver fibrosis collagen deposition * Aβ1-40 1 1 2 2 1 1 2 1 200032 2 200031 Aβ 1-40 120 CA1 Aβ 1-40 24h 4 Real-Time PCR CDK6 Bax Caspase-3 ELISA Tau Aβ 1-40 2 CDK6 Bax Caspase-3 Tau 1. 2g 2. 4g 4. 8g /kg Aβ 1-40 Tau CDK6 Bax Caspase-3 Aβ 1-40 Tau CDK6 Bax caspase3 Aβ 1-40 Alzheimer disease AD 1. 1 Aβ 1-40 2010 B 1 * 10YZ58 10DZ1971200 81072789 973 2010CB530400