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目的探究传统中药茜草水溶性提取物(RCAE)对高脂饮食诱导的肥胖大鼠体质量、脂肪含量及糖脂代谢指标的影响及机制。方法构建含PPARγ2启动子序列的p GL3-Enhancer-PPARγ2(625 bp)-Luc荧光素酶报告基因表达质粒;建立稳定转染该质粒的3T3-L1前脂肪细胞系;用不同浓度(0.1 mg/L~1 000 mg/L)水提法提取的RCAE作用该细胞或用100 mg/L RCAE作用不同时间,检测PPARγ2启动子活性;用100 mg/L RCAE刺激人脂肪细胞并检测PPARγ2 mRNA表达;同时高脂饮食喂养大鼠,观察小和大剂量RCAE干预对血糖、血脂、胰岛素水平、体质量和内脏脂肪质量等的影响。结果 10 mg/L RCAE能促进3T3-L1细胞荧光素酶的表达,是对照组的1.43倍(P<0.01);当浓度达1 000 mg/L时,荧光素酶活性增加至对照组的3.24倍(P<0.01)。100 mg/L RCAE刺激3T3-L1细胞28 h,荧光素酶活性达最大值,是对照组的2.72倍(P<0.01);还能显著促进人脂肪细胞中PPARγ2 mRNA的表达,是对照组的2.27倍(P<0.01)。与高脂对照组相比,小剂量茜草组的空腹胰岛素水平及HOMA-IR显著降低,内脏脂肪质量明显减少(P<0.05)。结论小剂量RCAE能显著减轻高脂饮食诱导的肥胖大鼠的内脏脂肪质量和改善胰岛素抵抗。其作用机制可能与增强PPARγ2基因启动子活性和促进PPARγ2 mRNA表达有关。
Objective To investigate the effect and mechanism of RCAE on the body weight, fat content and glucose and lipid metabolism of obese rats induced by high fat diet. Methods The pGL3-Enhancer-PPARγ2 (625 bp) -Luc luciferase reporter gene expression plasmid containing PPARγ2 promoter was constructed. The 3T3-L1 preadipocyte cell line stably transfected with the plasmid was established. L ~ 1 000 mg / L). The cells were treated with 100 mg / L RCAE for different time, the activity of PPARγ2 promoter was detected. Human adipocytes were stimulated with 100 mg / L RCAE and the expression of PPARγ2 mRNA was detected. At the same time, rats were fed on a high-fat diet to observe the effects of small and large doses of RCAE intervention on blood glucose, blood lipid, insulin level, body weight and visceral fat mass. Results 10 mg / L RCAE could promote the luciferase expression in 3T3-L1 cells by 1.43-fold (P <0.01), and at a concentration of 1 000 mg / L, luciferase activity increased to 3.24 Times (P <0.01). The luciferase activity of 3T3-L1 cells stimulated by 100 mg / L RCAE for 28 h reached the maximum, which was 2.72 times that of the control group (P <0.01), and also significantly promoted the expression of PPARγ2 mRNA in human adipocytes 2.27 times (P <0.01). Compared with the high-fat control group, the fasting insulin level and HOMA-IR in the low-dose Rubia cordyceps decreased significantly and the visceral fat mass decreased significantly (P <0.05). Conclusion Low dose RCAE can significantly reduce visceral fat mass and insulin resistance in obese rats induced by high fat diet. Its mechanism may be related to the enhancement of PPARγ2 gene promoter activity and PPARγ2 mRNA expression.