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1 Med J Chin PLA, Vol. 41, No. 12, December 1, GK [ ] GK 4 GK 32 ( 0.15g/kg) ( 1.3g/kg) ( 5.2g/kg) 8 8 Wistar (FINS) (GHb) (HOMA-IR) (IR) -1(IRS-1) 10 (P>0.05) (P<0.05)(P<0.05) (P<0.05) FINS GHb HOMA-IR (P<0.05) HOMA-IR (P<0.05) FINS GHb (P<0.05) IR IRS-1 (P<0.05) IR IRS-1 (P<0.05) GK IR IRS-1 [ ] GK [ ] R458.5 [ ] A [ ] (2016) [DOI] /j.issn Effect of Huangqi-Wumei granule on insulin resistance in GK rats LIU Bin 1, LI Shu-ling 2, LIU Kai 1, LI Ying-dong 1* 1 Institute of Integrated Traditional and Western Medicine, Gansu University of Chinese Medicine, Lanzhou , China 2 Provincial Level Key Laboratory, Affiliated Hospital of Gansu University of Chinese Medicine, Lanzhou , China * Corresponding author, lydj412@163.com This work was supported by the Research Project of Administration of Traditional Chinese Medicine of Gansu Province (GZK ) [Abstract] Objective To observe the effect of Huangqi-Wumei (Mongolian milkvetch-smoked plum, HW) granule on insulinresistanceingkrats.methods Thirty-two male 4-week-old GK rats were randomly divided into four groups (8 each): model group, positive control group (metformin hydrochloride enteric-coated tablets, 0.15g/kg), low dose HW granule group (HW granule 1.3g/kg) and high dose HW granule group (HW granule 5.2g/kg); meantime 8 male Wistar rats of eight-week-old were used as normal control group (normal saline). During 10 weeks of gavage, multiple measurements of rats' body weight and fasting blood glucose (FBG) were detected respectively on the 0, 2, 4, 6, 8 and 10 week. Fasting serum insulin (FINS) and glycosylated hemoglobin (GHb) were measured and insulin resistance index (HOMA-IR) was calculated at the end of the 10th week. Rats were sacrificed by acute blood loss, and the expressions of insulin receptor (IR) and insulin receptor substrate-1 (IRS-1) protein in rats' liverweremeasuredbywesternblotting.results Ten weeks after intervention, compared with the normal control group, no significant difference of the body weight existed in model group (P>0.05) and positive control group (P>0.05), but the body weight in HW granule groups reduced significantly (P<0.05). Compared with the normal control group, the FBG, GHb and HOMA-IR in the model group was significantly increased (P<0.05); compared with the model group, the HOMA-IR in HW granule groups and positive control group decreased significantly (P<0.05), the FINS and GHb in the high dose HW granule group and the positive control group decreased (P<0.05). Compared with the normal control group, the expression of IR and IRS-1 protein in the model group decreased (P<0.05). Compared with the model group, the expression of IR and IRS-1 protein in the HW granule groups [ ] (GZK ) [ ] [ ] ( ) ( ) [ ] lydj412@163.com
2 and the control group increased significantly (P<0.05). Conclusions HW granule may reduce the blood glucose, lose weight and attenuate insulin resistance of GK rats. The mechanism may be related to the increased expression of IR and IRS-1 protein in liver tissue of GK rats. [Key words] Huangqi-Wumei granule; rats, GK; insulin resistance 2 [1-2] (IR) -1(IRS-1) IRS-1 3 (PI3K) [3] ( IR/IRS-1 ) [4] 2 [5] GK IR/IRS SPF GK 32 ( SCXK( ) ) 4 SPF Wistar 8 ( SCXK( ) ) 8 SPF 12h 1.2 ( K K) (0.25g ) 1.3 ( Roche ) Infinite 200 PRO (Tecan ) TaKaRa BCA ( AK3501) AlphaImager 2200 ( Alpha ) PLCO ( Heraeus ) SDS-PAGE ( ) PowerPac Universal ( Bio-Rad ) IRS-1 ( GR GR ) GAPDH IgG( D ) ELISA 96T (GHb) ELIA 48T( A A) GK Wistar 8 ( : =5:2 ) 1.3g/kg( 30g 12g ) 5.2g/kg( 4 ) ( 4 3d )0.15g/ kg h h 10% (3ml/kg) 4ml (FINS) (GHb) HOMA (HOMA-IR) HOMAM- IR=FPG FINS/22.5 Western blotting IR IRS-1 60mg BCA SDS-PAGE PVDF ECL Image J 1.5 SPSS 17.0 x±s LSD-t
3 Med J Chin PLA, Vol. 41, No. 12, December 1, Tamhane's P< GK GK (P<0.05) GK (P>0.05) 10 (P>0.05) (P<0.05) (P>0.05) (P<0.05 1) 1 GK (g x±s n=8) Tab.1 Effect of Huangqi-Wumei granule on the body weight of GK rats (g, x±s, n=8) Time (week) Normal control group ± ± ± ± ± ±32.40 Model group ± ± ± ± ± ±11.03 Positive control group ± ± ± ± ± ±13.49 Low dose Huangqi-Wumei granule group ± ± ± ± ± ±26.92 High dose Huangqi-Wumei granule group ± ± ± ± ± ±24.59 P<0.05 compared with normal control group; P<0.05 compared with model group 2.2 GK 2 (P<0.05) (P<0.05) (P<0.05) 10 (P<0.05) (P<0.05 2) 2 GK (mmol/l x±s n=8) Tab.2 Effect of Huangqi-Wumei granule on fasting blood glucose of GK rats (mmol/l, x±s, n=8) Time (week) Normal control group 4.41± ± ± ± ± ±0.39 Model group 4.61± ± ± ± ± ±0.23 Positive control group 4.44± ± ± ± ± ±0.22 Low dose Huangqi-Wumei granule group 4.47± ± ± ± ± ±0.53 High dose Huangqi-Wumei granule group 4.34± ± ± ± ± ±0.41 P<0.05 compared with normal control group; P<0.05 compared with model group 2.3 GK FINS GHb HOMA-IR 10 FINS GHb HOMA-IR (P<0.05) HOMA-IR (P<0.05) FINS GHb (P<0.05 3) 2.4 GK IR IRS-1 Western blotting IR IRS-1 (P<0.05) IR IRS-1 (P<0.05 1) 3 GK FINS HOMA-IR GHb (x±s n=8) Tab.3 Effect of Huangqi-Wumei granule on FINS, HOMA-IR and GHb of GK rats (x±s, n=8) FINS (mu/l) GHb (ng/ml) HOMA-IR Normal control group 46.02± ± ±0.66 Model group 57.06± ± ±0.82 Positive control group 48.85± ± ±1.08 Low dose Huangqi-Wumei granule group 55.02± ± ±1.37 High dose Huangqi-Wumei granule group 49.41± ± ±0.99 P<0.05 compared with normal control group; P<0.05 compared with model group
4 IR GAPDH IRS-1 GAPDH RelativelevelofInsRprotein A B C D E 1 GK IR IRS-1 (x±s n=3) Fig.1 Effect of Huangqi-Wumei granule on the expressions of IR and IRS-1 protein in GK rats (x±s, n=3) A. Normal control group; B. Model group; C. Positive control group; D. Low dose Huangqi-Wumei granule group; E. High dose Huangqi- Wumei granule group. P<0.05 compared with normal control group; P<0.05 compared with model group Relative level of IRS-1 protein A B C D E 3 2 ( IR/IRS-1/PI3K/GLUT4 ) IR IRS [6-8] IR 2 2 [9] IRS IR IRS-1 IRS-2 IRS-2 IRS-1 IRS-1 [10-11] GHb [12] GK(Goto-kakizaki) Goto [13] Wistar GK Wistar 10%~30% Wistar 50%~55% [14] GK Wister GK IR [15] GK GHb(163.96±4.79ng/ml) (195.52±9.52ng/ml) (P<0.05) HOMA-IR (P<0.05) HOMA-IR (P<0.05) GK IR IRS-1 IR IRS-1 IR IRS-1 ( ) [16] 2
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