The Effects Of FFA In Insulin Resistance Of Adipose Tissue And The Application Of Solexa Sequencing On Detection Of MiRNAs In Animals

Insulin resistance (IR) is the condition in which normal amounts of insulin are inadequate to produce a normal insulin response from fat, muscle and liver cells. Insulin resistance in muscle cells reduces glucose uptake, whereas insulin resistance in liver cells results in impaired glycogen synthesis and a failure to suppress glucose production. Insulin resistance in fat cells reduces the effects of insulin and results in elevated hydrolysis of stored triglycerides in the absence of measures which either increase insulin sensitivity or which provide additional insulin. Increased mobilization of stored lipids in these cells elevates free fatty acids in the blood plasma. Obesity-induced chronic inflammation is a key component in the pathogenesis of insulin resistance and the Metabolic syndrome. Elevated plasma fatty acids are close related with insulin resistance induced by obesity. Here, we stimulate RAW264.7 macrophage with Free Fatty Acids(FFA), and then co-culcured them with 3T3-L1 adipocytes. We found that FFA weaken insulin sensibility of adipocyte by inducing secretion of TNF-a by macrophages.Illumina’s sequencing by synthesis (SBS) technology is the most successful and widely-adopted next-generation sequencing platform worldwide. This technology supports massively parallel sequencing using a proprietary reversible terminator-based method that enables detection of single bases as they are incorporated into growing DNA strands. A fluorescently-labeled terminator is imaged as each dNTP is added and then cleaved to allow incorporation of the next base. Since all four reversible terminator-bound dNTPs are present during each sequencing cycle, natural competition minimizes incorporation bias. The end result is true base-by-base sequencing that enables the industry’s most accurate data for a broad range of applications. A wide array of available sample preparation methods serve to enable diverse applications, including:whole-genome and candidate region resequencing, transcriptome analysis, small RNA discovery, methylation profiling, and genome-wide protein-nucleic acid interaction analysis.MicroRNAs (miRNAs), a class of about 22 nucleotide long non-coding RNAs derived from hairpin precursors, mediate the post-transcriptional silencing of an estimated 30% of protein-coding genes by pairing with complementary sites on the target mRNAs, thus blocking the translation or triggering the degradation of the target mRNAs. MiRNAs have been widely shown to modulate various critical biological processes, including differentiation, apoptosis, proliferation, the immune response and the maintenance of cell and tissue identity. In order to understand the role of miRNAs in gene regulation, an important way is the rapid and accurate quantitative detection the miRNA expression levels in various physiological and pathological station.Here, we detect the miRNAs expression profilings of sera and tissues of various animals using illumina’s sequencing by synthesis (SBS) technology. We report the surprising findings that not only endogenous animal miRNAs but also exogenous plant miRNAs are present in the sera and tissues of various animals. We also prove the conclusion with Q-PCR and Northern blotting.