| ObjectiveTo Investigate in vitro screening the adipocyte specific aptamer adipo8can combine with adipose tissue specifically in obesity mouse. To Investigate adipo8impact on adipogenic and the mechanism in obesity mouse preliminarily.Methods1.4weeks male C57BL/6mouse of86were randomly divided into two groups, standard diet group were12fed with standard and high fat group were74fed with standard diet2weeks and high-fat diet8weeks established Diet-Induced Obesity mouse model.2. Taking4obesity mouse the liver, kidney, muscle and epididymal adipose tissue made into two frozen sections using freezing microtome. Adipo8incubated, observed the tissue fluorescence signal using the upright fluorescence microscope and HE staining.4mouse received intraperitoneal injection partial phosphorothioate modified and with polyethylene glycol adipo8fluorescein labeled by cy3. Taking the liver, kidney, muscle and epididymal adipose tissue made into two frozen sections after injection of1h. Observed fluorescence signal and HE staining.3.24mouse divided into two groups and received intraperitoneal injection unmodified and modified adipo8fluorescein labeled by cy3seperately.2mouse were killed after injection of30min,1h,2h,4h,8h,24h. Taking the epididymal adipose tissue made into frozen section and imaged using the upright fluorescence microscope.4.32mouse divided into random sequence and adipo8groups. The mini-osmotic pumps embedded into mouse peritoneal. Continuous intraperitoneal infusion random sequence and adipo8respectively. Taking4mouse randomly, Retreated pump and weighed, calculated the lee’s index, measured FPG and TG1,2,3,4week later. Taking epididymal adipose tissue made into frozen section, observed adipocyte size using HE staining and statistical analysis. The level of PPARy expression in adipose tissue was detected by Western blotting.Results1. The weight, the lee’s index and FPG, TG of high-fat group were all higher than standard diet group(*P<0.05).2. Adipo8incubated,the result showed it was very strong fluorescence signal in adipose tissue while the liver, kidney, muscle were only weak signals under the upright fluorescence microscope. Intraperitoneal injection modified adipo8fluorescein labeled by cy31h showed fluorescence signal is strong in adipose tissue and others is weak.3. Unmodified adipo8combined with adipose tissue weak and short and modified adipo8was strong and long. The fluorescence signal of adipose tissue was first increase and then decrease. It was the strongest at1h and only weak signal at30min and24h.4. Continuous intraperitoneal infusion random sequence and adipo8, the weight, the lee’s index and FPG, TG in the first week was no significant difference between adipo8group and random sequence group(P>0.05). FPG and TG in adipo8group was all lower than random sequence group from the second to the forth week(*P<0.05).5. It showed that the adipocyte size was no significant difference between adipo8group and random sequence group in the first week(P>0.05)and it was smaller than random sequence group from the second to the forth week(*P<0.05) by using optical microscope with the same magnification single vision observation.6. The result of Western blotting showed that the expression of PPARy in adipose tissue of adipo8group and random sequence group were no difference in the first week (P>0.05) and adipo8group was lower than random sequence group from the second to the forth week (*P<0.05) The expression of PPARy in adipo8group decreased with the extension of time. Conclusions1. C57BL/6mouse feed with high fat diet8weeks established Diet-Induced Obesity mouse model.2. Adipo8can combine with white adipose tissue specifically in obesity mouse. Adipo8by partial phosphorothioate modified and with polyethylene glycol, enhances its binding with adipose tissue strength and extends the time.3. Adipo8can inhibit adipogenic in obesity mouse and the effect is time dependent in vivo. The mechanism of adipo8inhibition of adipogenic may be associate with PPARy path in obesity mouse. |
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