1.Establishment and stability of a hyperglycemic mouse model induced by alloxan
Lin CHEN ; Kai YUE ; Qin RU ; Xiang TIAN ; Qi XIONG ; Baomiao MA ; Lu LIU ; Rihui WU ; Junqiao XING ; Ning WANG ; Kun ZHANG ; Xiaowei ZHAO ; Wei CHEN ; Li HE ; Kangle OUYANG ; Yuanren SI ; Chaoying LI
Chinese Journal of Comparative Medicine 2014;(10):32-38
Objective To explore the influence of drug dosage, solvent and other main influencing factors on the successful establishment of alloxan-induced hyperglycemia mouse model and the effect on the stability of this model. Methods 160 6-8-week-old Kunming mice ofSPF grade, (male:female=1:1) were used in this study.The influences of different dosages of alloxan and solvent combinations on the successful establishment rate of the model, survival rate, body weight, fasting blood glucose, blood glucose area under curve, serum insulin level and their stabilities were dynamically observed for six weeks.Results By single intraperitoneal injection of 160 mg/kg bw alloxan ( pH 4.5 citrate sodium as solvent) , we were able to obtain a stable experimental hyperglycemic mouse model with higher levels of successful establishment rate (70%), survival rate (75%), fasting blood glucose (15-20 mmol/L), glucose area under the curve (55-65 mmol/L) and a lower but not loss of serum insulin levels (21 mIU/L).Conclusions In the present study we have carefully considered the influence of main factors such as drug dosages, solvent, etc., on the alloxan-induced experimental hyperglycemic mouse model, and successfully established this model after 6-week period observation of its stability.This model may provide a useful tool in the research of experimental diabetes and hypoglycemic functional studies.
2.Study on Improvement Effect of Methylated Urolithin A on Oleic Acid-induced Lipid Accumulation in Huh- 7 Cells and Its Me- chanism
Cong ZHANG ; Junxuan ZHOU ; Lei SHENG ; Junqiao MA ; Xin LI ; Guohua ZHENG ; Sidan LIU ; Zhengpeng QIU
China Pharmacy 2019;30(6):741-746
OBJECTIVE: To study the improvement effect and mechanism of methylated urolithin A on oleic acid-induced lipid accumulation in human liver cancer Huh-7 cells. METHODS: Oleic acid was adopted to induce lipid accumulation model cells. Huh-7 cells were divided into control group (culture medium), model group (1 mmol/L oleic acid), low-dose group (1 mmol/L oleic acid+10 μmol/L methylated urolithin A) and high-dose group (1 mmol/L oleic acid+20 μmol/L methylated urolithin A). Oil red O staining was used to observe lipid accumulation in cells. Triglyceride(TG) enzyme assay was applied to determine the TG content in cells. PCR was employed to detect the mRNA expression of FASN, SREBP-1, PPAR-α and PPAR-γ in cells. Western blotting was used to determine the protein expression of FASN in cells. RESULTS: After induced by oleic acid, a large amount of lipid droplet accumulated around the cells; the intracellular lipid and TG content, mRNA expression levels of FASN, SREBP-1 and PPAR-γ, protein expression levels of FASN were increased significantly, while mRNA expression level of PPAR-α was decreased significantly (P<0.01). After intervened with methylated urolithin A, lipid droplet around the cells decreased significantly; the contents of lipid and TG in cells were decreased significantly, while the mRNA expression levels of FASN, SREBP-1 and PPARγ and protein expression level of FASN were decreased significantly (P<0.05 or P<0.01). CONCLUSIONS: Methylated urolithin A can improve oleic acid-induced lipid accumulation in Huh-7 cells, the mechanism of which may be associated with inhibiting fat synthesis, promoting lipid metabolism and down-regulating the expression of metabolism-related factors as FASN, SREBP-1 and PPAR-γ.