| Objective and backgroundNon-alcoholic fatty liver disease (NAFLD) is a clinical syndrome with lesion of steatosis and fat storage in hepatic lobules but without alcohol abuse. Epideraiological survey shows that the prevalence of NAFLD has exceeded those of virus hepatitis and alcoholic liver disease (ALD), becoming a medical and social problem with common concern. In western country, the sickness rate is 2 to 3 times higher than hepatitis B, hepatitis C and ALD. In the last twenty years, the sickness rate in China and Asia Pacific region has raised about 7 to 40%; in Japan, increased 3 to 20 times and exceeded hepatitis C. Pathogenesis of NAFLD involve 5 interdependent patho-physiologic factors, namely abnormity of lipid metabolism, increase of ROS, lipid peroxidation in liver, stellate cell activation and abnormity of cytokines. The abnormity of cytokines in NAFLD pathogenesis is gaining more andmore regard in recent years. There are reports on cytokines(such as TNF and TGF -P 1) promoting effject on inflammation and f ibration of hepatic tissue, thus, NAFLD.But the cytokines with anti-inflammatory and anti-fibration properties areresearched few.Interleukin 10 (IL - 10) is an important anti-inflammatory, ant- fibration cytokines with immunoregulation effect. Its effect on endotoxin induced hepatic injury, virus hepatitis and fibration had been confirm with animal and human study. Then, does IL - 10 involve the pathogenesis of non-alcoholic fatty liver disease? In these study, rat NAFLD model was established with high fat diet, then the expression of IL - 10 mRNA in liver was analyzed with RT - PCR.Material and method19 healthy sexual matured SD rat, (weighting 260 - 280g, provide by animal center of medical college zhejiang university), Entered the 2 groups randomly: 10 in treatment group, feed on 88% common food plus 2% cholesterol and 10% lard; 9 in control group feed on common food .All rats were sacrificed after 4 weeks. The rats is weighted fastenly, get Blood 2ml from femoral vein to detect ALT and AST, anesthetize with 2% pentobarbital 30 - 40mg/kg intraperitoneal injection. The blood sample was taken, liver was weighed, hepatic tissue specimen was prepared with 10% formalin for paraffin section, some fresh specimen were stored in - 80 for RT - PCR.Detect ALT and AST: fresh Blood samples were Centrifugated at 3000rpm Velocity, serum was reserved and detected ALT and AST by automatic biochemistry analyzerHE dye: 60 1 hour dimethyl benzene dewax-gradient alcohol rehydration -wash with distal water hematoxyllin dye for 8minutes wash with distal water75% hydrochloric acid alcohol differentiation eosin dye 3 seconds wash with distal water-dimethyl benzene transparence and mount. Then observe under microscope.RNA extraction and reveres transcription: Total RNA was extract from liver tissue with Trizol, cDNA was transcripted using M - MLV reverse transcriptase kit ( GIBCOBRL), cDNA was kept at - 20癈. Take 2l cDNA as template, run PCR with primers. IL - 10 primers: 5' -ACT GGA GCA GGT GAA GAA TG- 3' and 5' -CCA GCC HA GGA TCG AAG TT-3', target fragment is 479bp. GAPDH was used as internal control: 5'-CAT GGT CTA CAT GTT CCA GT-3' and 5' -GGC TAA GCA GTT GGT GGT GC-3', target fragment 349bp.Mix 2ul template cDNA , 0.2 1 Taq enzyme, primers 1 1 each, 10mM dNTP 0.8 25mM Mgcl2 1.2 l ( end consistence 1.5mm) and 10X buffer 2 l, then add water to 20 . 94 denaturalization 5 minutes followed by 94 30 seconds 60 30 seconds 72 thirty seconds, for 30 cycles, then 725 minutes to stop reaction. 5 1 PCR product was add into 2% agarose gel (with 0. 5 g/ml bromize ingot) for electrophoresis, ultraviolet image was taken and optical density was measured.Statistic: Means is compared using student's t test.Results1. establishment of rat NAFLD model:1) general condition: All rats gain weight progressively, but those in treatment group more quickly. There is distinct swelling of liver in model group. The body weight, liver wet weight and ratio of liver weight to body weight of treat...
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