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侑朔玳靳
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1 19 7 LDL-C LDL-R 谢慧臣 1, 刘芬 1*, 田春漫 1, 向靖 2 2, 杨强 ( 1. 湖北民族学院医学院中西医结合教研室, 湖北恩施 ; 2. 湖北民族学院医学院附属医院中西医结合科, 湖北恩施 ) LDL LDL-R Wistar g kg g kg LDL Trizol RNA RT-PCR LDL-R mrna TC TG LDL-C B Apo B HDL-C A Apo A Apo A /Apo B P < P < T max LDL lag time LDL-R mrna P < P < LDL LDL-R mrna R doi /zgsyfjxzz A Effect of Jiangzhi Mixture on LDL Susceptibility to Oxidation and Liver Cells LDL-R Gene Expression of Hyperlipidemia Rats XIE Hui-chen 1 LIU Fen 1* TIAN Chun-man 1 XIANG Jing 2 YANG Qiang 2 1. Department of Integrated Traditional Chinese and Western Medicine Medicine School Hubei Nationalities College Enshi China 2. Department of Integrated Chinese and Western Medicine Affiliated Hospital Medical School Hubei Nationalities College Enshi China Abstract Objective To study the effect of Jiangzhi Mixture IM on low density lipoprotein LDL oxidative susceptibility and low density lipoprotein receptor LDL-R gene expression in rats with hyperlipidemia and elucidate the possible mechanism of IM regulating blood lipid. Method Sixty Wistar male rats were randomly divided into normal group model group simvastatin and IM high middle low dose groups 10 rats in each group. The rats in normal control group were fed with basal diet the rats in the other groups were fed with a high fat diet to establish hyperlipidemia rat models and during modeling the drugs were used for prevention. Physiological saline were given to the rats in control group and model group simvastatin group rats were given g kg - 1 simvastatin suspension solution and Jiangzhi Mixture low middle and high dose groups were intragastrically given ABA156 Tel xiehc_2004@ yeah. net * Tel liuflower813@ hotmail. com 245

2 g kg - 1 decoction respectively 10 ml kg - 1 once a day for 10 weeks during making model. After 10 weeks according to the method of abstracting eyeball blood the blood lipid spectrum of each individual animal in various groups were measured with full automatic biochemical analyzer. After separation of LDL its susceptibility to oxidation was determinated in vitro. According to the method of Trizol extracting liver RNA of rats LDL-R mrna expression was determinated by RT-PCR. Result Compared with model group the cholesterol TC triglycerides TG low density lipoprotein chelesterol LDL-C and apolipoprotein B Apo B levels in Jiangzhi Mixture groups were regulated and inhibited the high density lipoprotein chelesterol HDL-C apolipoprotein A Apo A levels and Apo A /Apo B ratio were increased meanwhile the differences were statistically significant P < or P < Compared with the model group the maximum rate of oxidation time T max and LDL oxidation lag time lag time were significantly prolonged and the relative expression levels of liver LDL-R mrna were significantly elevated in all Jiangzhi mixture dose groups and the differences were statistically significant P < or P < Conclusion Jiangzhi Mixture can modulate blood lipid metabolism in experimental hyperlipidemia rats models reduce LDL susceptibility to oxidation of hyperlipidemia rat significantly and induce liver tissue LDL-R mrna expression of hyperlipidemia rat and have good adjusting effect on lipid abnormalities. Key words Jiangzhi Mixture hyperlipidemia lipid profile LDL LDL-R gene hyperlipoidemia B Apo B Trizol Reagent Invitrogen Life Technologies L720R-3- HY-5F SLAN 12 PCR LDL- R mrna PCR LDL Invitrogen Biotechnology Co. LTD LDL-R ddh 2 O 2. 5 μmol L - 1 PCR β-actin 1 5'-CGTTGACATCCGTAAAGACCTC-3' 1. 1 Wistar 60 5'-TAGGAGCCAGGGCAGTAATCT-3' 10 ~ ± 20 g 517 bp LDL-R 5'-GCGAATGCTGC SYXK TCGTTGTG-3' 263 bp g 15 g 15 g 6 g 6 2% 5% 5% g 12 g 8 g 10 g 10% 0. 3% 0. 6% 77. 1% 1. 0 g ml TC TG TC TG LDL-C g L - 1 TC TG LDL-C HDL-C g ml A ApoA TAGTGATTGT-3' 5'-CCCTGAGAGTGC mmol L g ml - 1

3 LDL-C LDL-R 10 ml kg μl RNA RNA 2 PCR 2 μg RNA 1 μl oligo LDL 5 min 5 buffer 4 12 h 2 ml μl 10 mmol L - 1 dntps 2 μl RNAinhibitor 1μL r min min 1 μl PCR min TG TC LDL-C 80 5 min 3 PCR HDL-C Apo B Apo A 1 g L - 1 EDTA 0. 2 ml PCR 2 ml g 4 10 min 3 2 qpcr Mix μl 2. 5 μmmol L - 1 LDL 4 ph mmol L μl 2. 0 ddh 2 O 8. 5 PBS 24 h EDTA g L - 1 LDL dt 12 μlpcr 70 μl PCR 95 1 min CuSO 4 CuSO 4 LDL 15 s s s μmol L g L - 1 min s nm 15 min ΔΔCT A = CT A 180 min - CT B = CT CD 234 nm - CT K = A - B A 234 = 2 - K LDL CD LDL 2. 5 SPSS lag time x 珋 ± s P < T max LDL 4 LDL T max LDL lag time LDL-R 1 RNA ml Trizol Reagent TC TG LDL-C 100 mg 250 HDL-C P < μl g P < min 0. 8 TC LDL-C P < min g TC TG 10 min RNA LDL-C HDL-C P < g 5 min P < TG TC HDL-C LDL-C x 珋 ± s n = 10 mmol L - 1 /g kg - 1 TC TG HDL-C LDL-C ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± P < P < P < 0. P < Apo A Apo A /Apo B P Apo B Apo < P < Apo B A P < P < Apo A /Apo B Apo B Apo A Apo A / P < P < Apo B P < P <

4 Apo A Apo B Apo A /Apo B x 珋 ± s n = 10 /g kg - 1 Apo A /mmol L - 1 Apo B /mmol L - 1 Apo A /Apo B ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± LDL LDL Lag time T max P < P < LDL P < LDL P < P < ± LDL x 珋 ± s n = 10 /g kg - 1 Lag time T max ± ± ± ± ± ± min ± ± ± ± ± ± P < P < P < P < LDL-R mrna PCR LDL-R mrna P < P < TC TG LDL-C LDL-R mrna 8-10 P < P < β LDL-R mrna P < P < LDL-R mrna A x 珋 ± s n = 10 /g kg - 1 LDL-R mrna ± ± ± ± ± P < P < P < P < P <

5 3. J TG TC LDL-C HDL-C 4. Apo A /Apo B J LDL AS 5. J ox- 6. LDL AS J LDL 7. J. LDL LDL Lag J time T max LDL LDL-R J LDL 10. M. 90% LDL-R l. LDL-R 11. J. LDL-R LDL-R LDL-R 12. J J J LDL-C LDL-R J. J J LDL OX-LDL J

21 13 D / 2 10 ig ig 10 d 1. 0 g kg /2 2 β- 5 g kg g kg mg kg - 1 ig ig 24 h 5% 1 2% CMC-Na 2. 5 ml 30

21 13 D / 2 10 ig ig 10 d 1. 0 g kg /2 2 β- 5 g kg g kg mg kg - 1 ig ig 24 h 5% 1 2% CMC-Na 2. 5 ml 30 21 13 1 2 1* 2 1. 430065 2. 445000 20 10 5 g kg - 1 5 mg kg - 1 ig 10 d ig HE 1 TFF1 TFF1 P < 0. 01 P < 0. 01 P < 0. 01 P < 0. 05 R285. 5 doi 10. 13422 /j. cnki. syfjx. 2015130100 A 1005-9903 2015 13-0100-05