反应详情

Total cholesterol, triglyceride and glucose levels in the serum were measured twice for each using a commercially available kit (Bio Clinical system), and insulin level was measured by ELISA using a mouse insulin kit (Shibayaki, Japan). Lipids were extracted from the liver tissue according to Folch et al.'s method. After adding 1 mL of distilled water to 0.25 g of liver tissue, the liver tissue was homogenized using a Polytron homogenizer (IKA-Werke GmbH & Co., Ultra-Turrax, Staufen, Germany). After adding 5 mL of chloroform:methanol solution (2:1, v/v) to the homogenate and mixing well, the mixture was centrifuged at 1000×g for 10 minutes. After adding 2 mL of chloroform:methanol solution (2:1, v/v) again to the supernatant, the same procedure was repeated to completely separate the lipid components of the liver. After adding 3 mL of chloroform:methanol:0.05% CaCl2 (3:48:47, v/v/v) solution to the remaining pellets and mixing well for 1 minute, followed by centrifugation at 1000×g for 10 minutes, the resulting pellets were completely dried with nitrogen gas. The dried lipids were dissolved in 1 mL of methanol and then analyzed. The same kit (Bio Clinical system) as the one used for the serum analysis was used to measure the triglyceride level of the liver tissue.

1. extractionLipids were extracted from the liver tissue
2. additionAfter adding 1 mL of distilled water to 0.25 g of liver tissue
3. additionAfter adding 5 mL of chloroform
4. additionmethanol solution (2:1, v/v) to the homogenate and mixing well
5. additionAfter adding 2 mL of chloroform
6. customto completely separate the lipid components of the liver
7. additionAfter adding 3 mL of chloroform
8. additionmethanol:0.05% CaCl2 (3:48:47, v/v/v) solution to the remaining pellets and mixing well for 1 minute
9. waitg for 10 minutes
10. customthe resulting pellets were completely dried with nitrogen gas
11. dissolutionThe dried lipids were dissolved in 1 mL of methanol