OBJECTIVETo identify the association between PLIN 1237 polymorphism and obesity in Chinese Han adults.

METHODSA total of 994 adults (157 obese subjects, 322 overweight subjects, and 515 normal controls) were recruited from two rural communities. PLIN 1237 polymorphism was genotyped by polymerase chain reaction-restriction-fragment-length-polymorphism (PCR-RFLP). Association between PLIN polymorphisms and obesity status was estimated by ordinal logistic regression.

RESULTSThe three genotypes of PLIN 1237 were detected with a percentage of 54.3%, 37.1%, and 8.6% in TT, TC, and CC genotypes, respectively. For the PLIN 1237 polymorphism locus, the frequency of alleles T and C was 0.73 and 0.27, respectively. The PLIN 1237 polymorphisms were in Hardy-Weinberg equilibrium. PLIN 1237 polymorphism was not associated with obesity. The odds ratio for overweight or obesity for the CC+TC genotype was 0.8 (0.4, 1.4) in women (P = 0.4) and 0.6 (0.3, 1.3) in men (P = 0.2) after adjustment for age, education, household income and alcohol consumption, smoking, and physical activity.

CONCLUSIONChinese Han adults have a lower frequency of variant-allele C in PLIN 1237. PLIN 1237 T > C polymorphism is not significantly associated with obesity in northern Chinese adults.

Adult ; Asian Continental Ancestry Group ; genetics ; Carrier Proteins ; Female ; Genetic Predisposition to Disease ; Humans ; Male ; Obesity ; genetics ; Perilipin-1 ; Phosphoproteins ; genetics ; metabolism ; Polymorphism, Single Nucleotide ; genetics

Humans ; Diabetes Mellitus, Type 2/genetics* ; Hyperlipidemias ; Mutation ; Insulin Resistance/genetics* ; Metabolic Diseases ; Proto-Oncogene Proteins c-akt ; Perilipin-1/genetics*

We used CRISPR/Cas9 to delete plin1 of 3T3-L1 preadipocyte, to observe its effect on lipolysis in adipocytes and to explore regulatory pathways. We cultured 3T3-L1 preadipocytes, and the plin1 knockout vectors were transfected by electroporation. Puromycin culture was used to screen successfully transfected adipocytes, and survival rates were observed after transfection. The optimized "cocktail" method was used to differentiate 3T3-L1 preadipocytes. The glycerol and triglyceride contents were determined by enzymatic methods. The changes in lipid droplet form and size were observed by Oil red O staining. The protein expression of PLIN1, PPARγ, Fsp27, and lipases was measured by Western blotting. RT-PCR was used to measure the expression of PLIN1 and lipases mRNA. After the adipocytes in the control group were induced to differentiate, the quantity of tiny lipid droplets was decreased, and the quantity of unilocular lipid droplets was increased and arranged in a circle around the nucleus. Compared with the control group, the volume of unilocular lipid droplets decreased, and the quantity of tiny lipid droplets increased after induction of adipocytes in the knockout group. The expression of PLIN1 mRNA and protein in the adipocytes was significantly inhibited (P<0.05); glycerol levels increased significantly (0.098 4±0.007 6), TG levels decreased significantly (0.031 0±0.005 3); mRNA and protein expression of HSL and ATGL increased (P<0.05); PPARγ and Fsp27 expression unchanged in adipocytes. The above results indicate that the knockout of plin1 enhances the lipolysis of 3T3-L1 adipocytes by exposing lipids in lipid droplets and up-regulating lipases effects.
3T3-L1 Cells ; Adipocytes ; metabolism ; Animals ; CRISPR-Cas Systems ; Gene Knockout Techniques ; Lipase ; metabolism ; Lipolysis ; genetics ; Mice ; Perilipin-1 ; genetics ; metabolism

OBJECTIVETo observe the effect of coptis root extract (CRE) on the gene expressions of perilipin and peroxisome proliferator activated receptor gamma (PPAR-gamma) in atherosclerotic plaque of ApoE-gene knockout mice for exploring its plaque stabilizing action and possible mechanism.

METHODSThirty-three ApoE knockout mice, 6-8 weeks old, were fed with high-fat diet for 13 weeks. After mature atherosclerotic plaques being formed, the animals were randomly allocated into the control group, the CRE group, and the simvastatin group (as positive control) , 11 in each group. They were continuously fed with high-fat diet and to the two drug-treated groups, respective drugs in clinically recommended dose were given for another 13 weeks. Then all mice were sacrificed by the end of experiment. The morphology and composition of atherosclerotic plaques in 4 sections of aortic roots were examined with HE and Movat stain, the average number of fibrous caps buried in the plaque was observed and counted, and the gene expressions of perilipin and PPAR-gamma mRNA were determined by Real-time fluorescent quantitative PCR technology.

RESULTSAfter treatment for 13 weeks, the number of fibrous caps and the gene expression of perilipin mRNA in the CRE group was significantly lower (P<0.05), but gene of PPAR-gamma mRNA was higher (P<0.01) than those in the model group.

CONCLUSIONIn a clinically recommended dose, CRE can significantly decrease the frequency of plaque rupture in aorta of ApoE-gene knockout mice and do favour to plaque stability, its mechanism may be related to the promotion of PPAR-gamma mRNA expression and the inhibition of perilipin mRNA expression.

Animals ; Aorta ; pathology ; Apolipoproteins E ; deficiency ; genetics ; Atherosclerosis ; genetics ; pathology ; Carrier Proteins ; Coptis ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression Regulation ; drug effects ; Gene Knockout Techniques ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; PPAR gamma ; genetics ; Perilipin-1 ; Phosphoproteins ; genetics ; Plant Roots ; chemistry ; RNA, Messenger ; genetics ; metabolism ; Time Factors