Research On Effects Of KLF15in Porcine Preadipocytes Differentiation And Prediction Of Fat Deposition Capability

With the continuous improvement of living standards, more and more people payattention to improve the quality of pork, meat quality traits become an importantindicator of breeding evaluation. Fat between muscles has a significant impact toimprove the taste of pork products, flavor and nutritional value. This study is todetermine the effect of KLF15on procine pre-adipocytes differentiation and lipidosisand explore KLF15regulate glucose level then affect lipidosis. Provide a theoreticalbasis for regulating lipidosis from the molecular level, improving pork quality andselecting porcine breedings.This study, firstly, choose Junmu-1st6-day-old health piglets separating primaryporcine pre-adipocytes, culture and induction of differentiation, then at differentstages of differentiation by oil red O staining, detect the mRNA expression level ofadipose differentiation marker genes PPARγ, C/EBPα, AP2and KLF15by RT-PCR.Results indicated that the expression level of adipose differentiation marker genePPARγ and C/EBPα significant increase (P <0.01), AP2expression of relative rise (P<0.05); induced porcine pre-adipocytes after24h, KLF15expression levels increasedrapidly and reached the maximum value (P <0.01), followed by a downward trend, butstill higher than before the induction of the expression level.Subsequently, this study use siRNA inhibit KLF15mRNA levels inpre-adipocytes, and use of RT-PCR detect changes in the expression of adipocytedifferentiation marker genes PPARγ, C/EBPα, AP2, and KLF15associated geneFSTL1, FASN, SCD, p300and FATP1. The results proved that the expression ofPPARγ, C/EBPα, AP2, p300and FATP1significantly reduced (P <0.01), expression ofFSTL1, FASN and SCD had no significant change. Meanwhile, after the inhibition ofmRNA expression of KLF15, triglyceride levels of inhibiting group decreasedsignificantly compared with the control group (P <0.05), lipid droplets significantlyreduced and smaller.Based on these findings, this study detect the content of KLF15，GLU, CHO and TG of different months of lean type(Junmu-1st) and fat type(Songliao blackswine)castrated boars serum by oxidase method detection and ELISA, and explore themethod for predict the lipidosis capability. The results showed that GLU levels of3and6months Songliao Black Swine and Junmu-1st are5.03±0.44mmol/L,5.25±0.55mmol/L,4.73±1.25mmol/L and3.45±0.15mmol/L. The CHO levels of3and6months Songliao Black Swine and Junmu-1st are1.57±0.41mmol/L,1.15±0.25mmol/L,2.10±0.18mmol/L and1.63±0.39mmol/L. The TG levels of3and6months Songliao Black Swine and Junmu-1st are0.16±0.02mmol/L,0.33±0.19mmol/L,0.44±0.19mmol/L and0.26±0.07mmol/L. The correlation coefficientsof KLF15with GLU, CHO, TG respectively are0.8010,0.0263,0.6299. The contentof KLF15in procine serum has a significant positive correlation with glucose levels,and it also has a positive correlation with triglycerides levels.These results illustrate that KLF15can regulate glucose levels and involve inthe regulation of pre-adipocytes differentiation, and also can affect adipocytestriglyceride synthesis and lipidosis. In the practical production, we can indirectestimate KLF15content and predict the capacity of procine lipidosis through detectglucose levels. This way can understand the role of KLF15in procine lipidosis and asa molecular marker to provide a theoretical basis for procine meat qualityimprovement and the early selection and breeding.