BACKGROUND Myeloid-related protein 8/14 (MRP8/14) is secreted by macrophages and formed by MRP8 and MRP14, which is closely related to vascular inflammation. Chronic vascular inflammation plays a significant role in the development and progression of diabetic kidney disease (DKD). This study aims to investigate the relationship between MRP8/14 and DKD. METHODS A total of 80 individuals with type 2 diabetes were divided into four groups, according to the baseline urinary albumin/creatinine ratio (ACR) levels Serum concentrations of MRP8/14 were measured by ELISA. The clinical variables were obtained through physical examination, illness history, or laboratory evidence. RESULTS As DKD worsened, the level of serum MRP8/14 increased gradually, and MRP8/14 has a significantly positive correlation with ACR (r = 0.349, P = 0.002), body mass index (BMI) (r = 0.288, P = 0.009), serum creatinine (Cre) (r = 0.392, P < 0.001), blood urine nitrogen (BUN) (r = 0.333, P = 0.003), systolic blood pressure (SBP) (r = 0.301, P = 0.007), and a negative correlation with the estimated glomerular filtration rate (eGFR) (r = -0.478, P < 0.001). Logistic regression analysis showed that age, Cre, eGFR, ACR, and MRP8/14 were associated with the progression of DKD (P < 0.05). CONCLUSIONS The serum MRP8/14 is correlated significantly with the progression of DKD, suggesting that MRP8/14 may be an independent predictor of the progression of DKD.
The proliferation and differentiation of preadipocytes are regulated by microRNAs (miRNAs), hormones and other factors. This study aimed to investigate the effects of miR-331-3p on the proliferationand differentiation of preadipocytes in addition to fatty acid metabolism. The data indicated that miR-331-3p is a novel regulator of cellular differentiation. It was observed that miR-331-3p was capable of inhibiting cellular proliferation. Furthermore, miR-331-3p was highly expressed during cellular differentiation andappeared to promote the process. In addition, dual fluorescein analysis showed that dihydrolipoamideS-succinyltransferase (DLST) is a target gene of miR-331-3p, and over-expression of miR-331-3p could regulate the metabolism of fatty acids in the citrate pyruvate cycle by targeting DLST expression. In summary, these findings indicated that miR-331-3p exerts contrasting effects on the processes ofproliferation and differentiation of preadipocytes.
The activity of inducible nitric oxide synthase(iNOS) in the liver,the concentration of nitric oxide(NO) in the serum,the coccidian oosysts per gram feces(OPG)and anatomic structures of rabiits' livers and micoscopic pathological changes were studied.The rabbits infected with Eimeria stiedai were injected with the substrate of nitric oxide synthase—L-arginine(L-Arg) with different concentrations and the inhibitor of inducible nitric oxide synthase-L-aminoguanidine(L-AG) through the abdominal cavity.The results showed that after infected with Eimeria stiedai,the activity of inducible nitric oxide synthase of rabbits' liver chips and the concentration of nitric oxide in the serum increased gradually.The control group,L-arginine group,and L-aminoguanidine group reached the top after infected 12 days,and then decreased gradually and reached the level before infected at about 23days;L-arginine could enhance the activity of inducible nitric oxide synthase in the liver and make it release large amount of nitric oxide,inhibit or kill Eimeria stiedai and ease the pathologies caused by Eimeria stiedai;On the contrary,L-aminoguanidine inhibits inducible nitric oxide synthase in the liver and reduces nitric oxide produced,so that the inhibition on Eimeria stiedai was weakened or even not existed andaggravated the pathologies.The results implied that nitric oxide and inducible nitric oxide synthase indeed participate in the process of rabbits' infection with Eimeria stiedai and nitric oxide played the definite role of inhibition or killness on coccidia of rabbites.
To investigate the effect of miR-331-3p on the proliferation of porcine renal epithelial cells (PK15) and its mechanism, the pcDNA 3.1(+) overexpression vector of miRNA-331-3p (pcDNA 3.1(+)-miR-331-3p) was constructed. PK15 cells were divided into four groups, including experimental group, experimental control group, inhibitor group and inhibitor control group. Experimental group and experimental control group were transfected with pcDNA 3.1(+)-miR-331-3p and pcDNA 3.1(+), respectively. Inhibitor group and inhibitor control group were transfected with miR-331-3p inhibitor and miR-331-3p negative control (miR-331-3p NC), respectively. Above all, CCK-8 reagent was used to plot the cell proliferation curve and Propidium (PI) staining was used to detect the proportion of cell stages. Secondly, its expression change were detected by quantitative real-time PCR that included the growth inhibitory protein family member 5 (ING5), cyclin-dependent kinase 2 (CDK2), cyclin-dependent kinase 3 (CDK3), cyclin-dependent kinase 4 (CDK4), Cyclin B and cyclin-dependent kinase inhibitor 1A (CDKN1A). The results showed that the expression of miRNA-331-3p was significantly increased in the experimental group. The cell proliferation curve showed that the number of cells in experimental group was significantly higher than that in experimental control group or inhibitor control group at 48 h and 72 h (P<0.05). Simultaneously, Inhibitor group was significantly lower than experimental control group or inhibitor control group in the number of cells at 48 h and 72 h (P<0.05), but there was no significant difference between the experimental group and the control group. Compared with the experimental control group, the proportion of cells of experimental group in G0/G1 phase decreased, the proportion of S phase and G2/M phase increased, and the inhibitor control group showed the opposite trend. Simultaneously, the expression levels of CDK2, CDK3, CDK4 and Cyclin B genes in the experimental group were significantly increased, while ING5 and CDKN1A genes inhibiting proliferation showed a significant downward trend. These results demonstrate that the miR-331-3p overexpression vector was successfully constructed, and miR-331-3p has the ability to promote the proliferation of PK15 cells. The study lays a solid foundation for further research for its role in pig growth and development.本研究旨在阐明猪miR-331-3p 对细胞增殖的影响，探讨其对细胞增殖的作用机制首先构建了miR-331-3p 的过表达载体pcDNA 3.1 (+)-miR-331-3p，并将将PK15 细胞分为4 组，分别为实验组、实验组对照组、抑制剂组和抑制剂对照组。实验组和对照组分别转染pcDNA 3.1(+)-miR-331-3p 和pcDNA 3.1(+)。抑制剂组和抑制剂对照组分别转染miR-331-3p Inhibitor 和miR-331-3p 阴性对照(miR-331-3p NC)。通过在各组添加CCK-8试剂绘制细胞增殖曲线，并使用PI 染色检测细胞所处周期比例。同时，利用实时荧光定量PCR (Quantitative real-time PCR，qPCR) 检测生长抑制蛋白家族成员5 (Inhibitor of growth family member 5，ING5)、细胞周期蛋白依赖性激酶2 (Cyclin dependent kinase 2，CDK2)、细胞周期蛋白依赖性激酶3 (Cyclin dependent kinase 3，CDK3)、细胞周期蛋白依赖性激酶4 (Cyclin dependent kinase 4，CDK4)、细胞周期蛋白B (Cyclin B) 和细胞周期蛋白依赖性激酶抑制剂1A (Cyclin dependent kinase inhibitor 1A，CDKN1A) 的表达变化。结果表明，实验组miR-331-3p表达量显著升高，细胞增殖曲线表明48 h 和72 h 时细胞数目均呈现出实验组>实验对照组和抑制剂对照组>抑制剂组的趋势 (P<0.05)。与实验对照组相比，实验组处于G0/G1 期的细胞比例下调，S 期和G2/M 细胞的比例上调，抑制剂对照组趋势与之相反；同时，实验组中与促进增殖的基因CDK2、CDK3、CDK4 和Cyclin B 的mRNA 表达水平均显著升高，而抑制增殖的基因ING5 和CDKN1A 均表现出显著下降的趋势。本研究成功构建了miR-331-3p过表达载体，且发现miR-331-3p 具有促进猪肾上皮细胞增殖的能力，研究结果为深入研究miR-331-3p 在猪生长发育中的作用机制奠定了基础。.
To explore the relationship between the expression of PID1 gene and fat deposition, we cloned CDS of PID1 from porcine fat and muscle tissues by RT-PCR using degenerate primers, and investigated expression of this gene in various tissues (i.e., liver, backfat, and muscle tissues) of different breeds (i.e., Yorkshire, Laiwu, and Lulai Black) by real-time fluorescence quantitative PCR. The results showed that 654 bp CDS of porcine PID1 was obtained by sequencing and was 93.88%, 66.94% and 88.07% identical to those of the human, rat, and Bos taurus, respectively. The expression of PID1 mRNA in various tissues and breeds, on the whole, tended to be liver > fat > muscle and Laiwu > Lulai Black > Yorkshire, respectively. For different breeds, PID1 mRNA abundance in liver had significant difference (P < 0.05), but had no significant differences in fat and muscle tissues between Laiwu and Lulai Black (P > 0.05). For the three groups of Laiwu pigs with high (LWH), intermediate (LWI), and low IMF content (LWL), PID1 mRNA level was higher in liver tissue of LWH than that of LWL significantly (P < 0.05), and was higher in muscle tissue of LWH than that of LWI and LWL significantly (P < 0.05). PID1 mRNA abundance was not correlated with IMF in these three tissues of Laiwu breed, but it was positively correlated with IMF in the tissues of these three breeds (P < 0.05). These results implied that the expression of PID1 may be related to fat deposition.
The proliferation and differentiation of preadipocytes are regulated by microRNAs (miRNAs), hormones, and other factors. This study aimed to investigate the effects of miR-331-3p on the proliferation and differentiation of preadipocytes in addition to fatty acid metabolism.Preadipocytes were transfected with miR-331-3p mimics, miR-NC, or miR-331-3p inhibitor to explore its effect on cell proliferation and fatty acid accumulation. Furthermore, preadipocytes were transfected with pre-miR-331-3p, pcDNA3.1(+), or miR-331-3p inhibitor to explore its effect on differentiation.It was observed that miR-331-3p could inhibit preadipocytes proliferation. Furthermore, miR-331-3p was highly expressed during cellular differentiation and appeared to promote the process. In addition, dual fluorescein analysis showed that dihydrolipoamide S-succinyltransferase (DLST) is a target gene of miR-331-3p, and overexpression of miR-331-3p could regulate the metabolism of fatty acids in the citrate pyruvate cycle by targeting DLST expression.In summary, these findings indicated that miR-331-3p exerts contrasting effects on the processes of fat deposition.
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