lncRNA-7896对熊果苷诱导黑色素沉积受阻淅川乌骨鸡胸肌、腿肌的影响OA

The study aimed to construct a melanin deposition obstruction model by exogenous addition of arbutin,and study the effects of lncRNA-7896 on melanin deposition-related genes,tyrosinase activity and melanin content in Xichuan Black-bone chickens.Eighty breeding eggs of Xichuan Black-bone chicken were selected and randomly divided into 4 groups.The control group(CON)was injected with 4 μL of PBS solution,the arbutin treatment group(ARB)was injected with 4 μL of 2 μmol·L-1 arbutin,the empty vector control group(A-NO)was injected with 4 μL of 2 μmol·L-1 arbutin and the empty vector of transposon,and the lncRNA-7896 overexpression group(A-LNC)was injected with 4 μL 2 μmol·L-1 arbutin and transposon lncRNA-7896 vector(transposon vector:transposon coenzyme vector was 2∶1).The test period was 28 d,starting from the hatching of the breeding eggs to the 7th embryo age,and the test period was 21 d.After the chicks hatched and were raised for one week,samples of heart,liver,pectoral muscle and leg muscle of Xichuan Black-bone chickens were collected.The collected tissue samples were extracted with RNA and reverse transcribed into cDNA.The expression levels of lncRNA-7896,MITF,TYR,and TYRP-1 were detected by real-time fluorescence quantitative PCR(qRT-PCR).Meanwhile,the tyrosinase activity in pectoral muscles and leg muscles was detected by ELISA kits.The melanin content in the pectoral and leg muscles was detected by the colorimetric reagent kit.The melanin content in the pectoral and leg muscles was visualized by the Masson-Fontana staining reagent kit.Finally,the data were statistically analyzed by one-way analysis of variance using SPSS 23.0 software.The results showed that,compared with the control group,the addition of 4 μL 2 μmol·L-1 arbutin significantly downregulated the expression levels of MITF,TYR and TYRP-1 genes(P<0.05);Meanwhile,the activities of tyrosinase and melanin contents in pectoral muscle and leg muscle were also markedly decreased after 4 μL 2 μmol·L-1 arbutin treatment(P<0.05).Notably,exogenous introduction of lncRNA-7896 in Xichuan Black-bone chickens with inhibited melanin deposition could distinctly upregulate the expression of MITF,TYR and TYRP-1 genes,and effectively increase tyrosinase activity as well as melanin content in pectoral and leg muscles(P<0.05).In summary,arbutin can suppress melanin deposition in Xichuan Black-bone chickens,whereas lncRNA-7896 can significantly restore the expression of melanogenic genes and increases both tyrosinase activity and melanin content in the pectoral and leg muscles after the deposition has been blocked.