1.Investigation on the role of endoplasmie reticulum stress in the fatty acids-induced hepatocyte injury
Jia DING ; Guowen TANG ; Zhuping FAN ; Dekai QIU
Chinese Journal of Digestion 2011;31(11):729-733
ObjectiveTo explore the role of fatty acid on endoplasmic reticulum (ER) stress and to detect the influence of tauroursodeoxycholic acid (TUDCA) on the expression of ER related genes in steatosis hepatocytes by establishing different models of hepatic steatosis.MethodsHealthy adult hepatocyte cell line HL-7702 was taken as research object.The model of hepatic steatosis was established with palmitic acid alone or mixed fatty acids.The cell viability was measured and calculated through CCK-8 positive cell proliferation assay.The cell morphology and steatosis was observed.The intracellular triglyceride content was tested with the triglyceride determination kit.With TUDCA intervention,the relative expression of Glucose-regulated protein 78 (GRP78) and CCAAT/Enhancer binding protein homologous protein (CHOP) at mRNA level was determined by real-time PCR.ResultsThe viability of hepatocyte was influenced once the concentration of mixed fatty acid reached 0.5 mmol/L or palmitic acid was 0.125 mmol/L.The effect of palmitic acid alone was stronger than that of mixed fatty acid on hepatocyte injury.The content of intra-hepatocyte triglyceride gradually increased in mixed fatty acid group,which only significantly increased after treated for 12 hours in palmitic acid alone group and then there was no significant change.There was no significant difference in relative expression of GRP78 and CHOP at mRNA level in various concentrations and treated time of mixed fatty acid group.After treated with palmitic acid at 0.5 mmol/L,intra-hepatocyte relative expression of CHOP at mRNA level increased obviously,however there was no effect on GRP78mRNA expression.After treated with palmitic acid at 1.0 mmol/L,both intra-hepatocyte GRP78 and CHOP mRNA relative expression increased.There was no significant difference in GRP78 and CHOP mRNA relative expression before and after ER stress in TUDCA intervented low dose palmitic acid group.There was significant difference in CHOP mRNA relative expression before and after ER stress in TUDCA intervented high dose palmitic acid group (at 12hrs:8.6400 to 5.1032; at 24hrs:13.7948to 6.4928,P=0.042 and 0.017),while no significant difference in GRP78 mRNA expression.ConclusionAt same fatty acid concentration,the larger propotion palmitic acid has,the more severe injury hepatocyte has.The regulation of palmitic acid on intra-hepatic ER stress is a time and dose dependent manner.TUDCA may improve palmitic acid induced ER stress to some degree.
2.FEATURES OF SURFACE CELLS OF MESENTERY UNDER SCANNING ELECTRON MICROSCOPE
Haobai ZHOU ; Teng ZHOU ; Qin LU ; Zhuping TANG ; Changyan WU ;
Acta Anatomica Sinica 1955;0(03):-
This paper reports a simple method using the mesentery as a naturai mountingsubstrate for the study of mesenteric free cells under the scanning electron micros-cope.Mouse mesentery was cut into pieces about Ⅰ square centimeter in size andadhered on to neutral filter paper.The specimen were fixed in 5% glutaraldehyde and1% osmium tetroxide for 20 minutes,each step washed lightly with pH 7.4 PBS,dehydrated with ascending series of alcohol,put into amyl acetate,critical pointdried with CO_2,coated with gold and examined in scanning electron microscope.The morphologic features of free cells in the peritoneal lumen,such as polymorpho-nuclear leucocytes,macrophages and lymphocytes could be observed satisfactorily.We studied in particular the relationship between immunoreactive cells and targetcells(fetal liver cell)injected into the peritoneal cavity,Polymorphonuclear leuco-cytes could be seen invading into and eroding the target cells and macrophage cau-sing their swelling and lysis.Cooperative action of immunoreactive cells and anincrease in number of milk spots were also observed.
3.Study on Antitumor Mechanism of Periplaneta americana Extract C Ⅱ-3 on MFC Tumor-bearing Mice Based on 1H-NMR Metabonomics
Ying LI ; Zhuping TAO ; Xu CHANG ; Hongli OU ; Lun JIANG ; Canwei LI ; Yanlong TANG ; Yue ZHOU ; Li BAI ; Pengfei GAO
China Pharmacy 2020;31(12):1446-1451
OBJECTIVE:To preliminarily study the antitumor mechanism of Periplaneta americana extract C Ⅱ-3 on MFC tumor-bearing mice. METHODS :Balb/c mice were randomly divided into model group (normal saline 20 mL/kg)and C Ⅱ-3 group (200 mg/kg),with 6 mice in each group. MFC cell suspension (0.2 mL)was injected under the right armpit of mice. On the next day,mice were given relevant medicine intragastrically ,once a day ,for consecutive 10 d. 24 h after the last administration ,Based on the measurement of tumor size , 1H-NMR technology combined with unsupervised PCA ,supervised PLS-DA and OPLS-DA were used to compare metabolic spectrum of liver tissue from tumor-bearing mice of 2 groups,to analyze differential metabolites and to explore the potential antitumor mechanis m of C Ⅱ -3. RESULTS :Compared with model group ,the tumor body was significantly reduced in tumor-bearing mice of C Ⅱ-3 group. There were differences in 1H-NMR spectra between the 2 No.81960712); groups. According to unsupervised PCA ,supervised PLS-DA and OPLS-DA ,totally six potential differential metabolites ,as glycogen (increased),pyruvate (decreased),arginine (de- creased),hydroxyproline (increased),inosine (increased) and niacinamide (increased),were identified in the liver tissue,which were mainly attributed to the metabolism of arginine ,energy and nucleic acid. CONCLUSIONS:The anti tumor effect of C Ⅱ-3 may be related to the regulation of arginine metabolism ,energy metabolism and nucleic acid metabolism.