1.Expression levels of GLUTs and SIRTs in diabetic liver lesions
Yu GUO ; Wenfan BAI ; Yaping TIAN ; Feiyang LUO ; Shuyuan JIA ; Mingxiu LUO ; Qing YAO
Chinese Journal of Pathophysiology 2024;40(2):326-334
AIM:To study the expression of glucose transporters(GLUTs)and silent information regulators(SIRTs/sirtuins)in the liver of diabetic rats and human hepatocytes(LO2 cells)treated with high glucose.METHODS:(1)Twenty male SD rats were randomly divided into normal control(NC)group and diabetes mellitus(DM)group.The rats in DM group were given single intraperitoneal injection of streptozotocin(STZ,60 mg/kg)to establish the DM model,while the rats in NC group were intraperitoneally injected with equal volume of solvent once.Fasting blood glucose(FBG)and body mass were measured every 2 weeks.After 12 weeks of rearing,the blood and liver tissues of the rats were ob-tained after anesthesia with 1%sodium pentobarbitone,the biochemical indicators of blood were detected,and the liver in-dex was calculated.Hematoxylin-eosin(HE)staining and periodic acid-Schiff(PAS)staining were used to observe liver histopathological changes.Lipid accumulation in liver tissues was detected by oil red O staining.The expression levels of GLUTs and SIRTs family member proteins were detected in rat liver tissues.(2)The LO2 cells were treated with different concentrations of glucose for 48 h.The viability of the cells in each group was measured by CCK-8 assay,and Western blot was used to detected the protein expression levels of GLUTs and SIRTs in the cells.RESULTS:(1)Compared with NC group,the rats in DM group were depressed,lost weight,and the FBG and liver index were significantly increased(P<0.05).The results of HE staining showed that the hepatic sinuses were dilatated and congested near the central vein in DM rats,and mild edema and scattered infiltration of inflammatory cells were found in liver cells.The results of oil red O staining showed the red fat droplets were diffusely scattered within liver cells in DM group.The results of PAS staining showed that there were numerous diffuse light purple circular droplets in the cytoplasm of the liver cells in the central ve-nous area of the DM rats.Western blot showed that the protein levels of GLUTs were higher and the protein levels of SIRTs were lower than those in NC group(P<0.01).(2)The results of CCK-8 assay showed that the viability of LO2 cells was increased in 50 mmol/L glucose group(P<0.01),without significant difference in 75,100 and 125 mmol/L glucose groups(all P>0.05),and decreased in 150,175 and 200 mmol/L glucose groups(all P<0.01).Later,150 mmol/L glu-cose was used as the high-glucose intervention condition.Western blot showed that the protein levels of GLUTs and SIRTs in LO2 cells under high glucose intervention were consistented with the results in animal experiments.CONCLUSION:High concentration of glucose can cause liver damage in SD rats and reduce the viability of human hepatocytes(LO2 cells).It can also increase the expression of GLUTs and decrease the expression of SIRTs in rat liver tissues and LO2 cells.Therefore,GLUTs and SIRTs family members may be the target proteins of diabetes-induced liver injury.
2.Characteristics Evaluation and Application Analysis on Animal Models of Recurrent Spontaneous Abortion
Tiansong DING ; Jinghong XIE ; Bin YANG ; Heqiao LI ; Yizhuo QIAO ; Xinru CHEN ; Wenfan TIAN ; Jiapei LI ; Wanyi ZHANG ; Fanxuan LI
Laboratory Animal and Comparative Medicine 2024;44(4):393-404
Objective To summarize and evaluate the characteristics of current recurrent spontaneous abortion(RSA)animal models at home and abroad,and to provide reference and guidance for the standardized preparation of RSA models.Methods"Recurrent spontaneous abortion"and"animal model"were used as co-keywords in CNKI,Wanfang,VIP,PubMed and Web of Science databases to search the RSA animal experimental literature,covering the period up to January 20,2024,and a total of 1 411 articles were collected.The analysis focused on construction methods and essential elements of RSA animal models,the modeling process and result evaluation,as well as the application of these models in pharmacological and pharmacodynamic research.An Excel table was established for systematic analysis and discussion.Results A total of 138 experimental studies were obtained after screening.In constructing RSA animal models,immunological models were the most widely used in Western medicine(96.92%),with the Clark model being the main one(92.31%).In traditional Chinese medicine(TCM)models,70.00%were kidney deficiency-luteal inhibition-syndrome combination models,20.00%were kidney deficiency and blood stasis models,and 10.00%were deficiency-heat syndrome models.Most animals were selected at 6-8 weeks(33.86%)and 8 weeks(32.28%)of age.The majority of animals were paired for mating at 18:00 on the day of cage pairing.In 81.03%of literatures,vaginal plugs were checked once the following morning,with 8:00 being the most common time(17.02%).The most commonly used drug administration cycle was 14 days of continuous gavage after pregnancy.Among the tested drugs,Western drugs were mainly protein-based(29.17%),while TCM drugs were mainly TCM decoction(81.11%).The most frequently used methods for detecting indicators included visual observation of embryos(22.54%),western blot(15.96%),PCR(13.58%),ELISA(12.91%),HE staining(10.80%)and immunohistochemistry(9.39%).Conclusion The etiology of RSA is complex,and corresponding animal models should be established based on different etiologies.Clark model is commonly used in the construction of Western medicine model,while the kidney deficiency-luteal inhibition-syndrome combination model is predominant in TCM.RSA animal model is widely used in related research,but systematic evaluation needs to be strengthened.