The Correlation Study Between 43ku ES Antigen Of Trichinella Spiralis And The Formation Of Nurse Cells

Trichinella spiralis is an intracellular parasitic nematode that infects in skeletal muscle cells, and the infected muscle cells would lost its intrinsic organization character and turn to a new tissue structure, Nurse cells. It was believed that excretory-secretory (ES) proteins of Trichinella spiralis are involved in the process of the formation of Nurse cells, and Trichinella spiralis 43ku ES antigen is one of them.The study established a assay method to detection expressions for Trichinella spiralis 43ku ES antigen gene (Ts43) and regulatory factors MyoD and Id1 gene of mice skeletal muscle cells with a real-time fluorescence quantitative PCR (RQ-PCR). To explore in depth the relationship between the level of Ts43 gene expression and the formation of nurse cells, and the information obtained will provide the basis for further studies on the relationship between the level of Ts43 gene expression and the formation of nurse cells.According to the known sequence of Ts43 gene and 18S rRNA gene of Trichinella spiralis published in GenBank, the primers and probes was designed. With the reference gene, the housekeeping gene (18S rRNA), the total RNA of Trichinella spiralis was dealt with normalization, and the expression of Ts43 gene was calculated with standard curve method. The method was utilized for detecting Ts43 gene dynamic expression in Trichinella spiralis at different parasitism periods (include: adult worms, newborn larva, pre-encysted larva and muscle larva). The results showed that Ts43 gene was expressed in the whole parasitism period, but the Ts43 gene expression level of adult and newborn larva was very low compared with the other parasitic worm. The gene expression at 18 day post-infection (p.i.) began to rise, at 30 day p.i. reached a peak and then began a gradual decline, but at 58 day p.i. Ts43 gene expression level was still higher than adult worm and newborn larva.According to the sequences of regulatory factors MyoD, Id1 and G3PDH gene of mice skeletal muscle cells published in GenBank, primers was designed. With the reference gene, the housekeeping gene (G3PDH), the total RNA of mice skeletal muscle cells was dealt with normalization, and the MyoD and Id1 gene expressions was calculated withâ–³Ct value. The method was utilized for detection of MyoD and Id1 gene dynamic expression in mice skeletal muscle, which are at Trichinella spiralis different parasitism periods (7 day, 14 day, 18 day, 22 day, 26 day, 30 day, 34 day, 38 day, 48 day and 58 day p.i.)and before and after vivo transfection of Ts43 gene. The results of study showed that MyoD, Id1 gene expression continued to increase in the mice skeletal muscle cells at the beginning of Trichinella spiralis infection. The level of MyoD gene expression was at the highest value at 26 day p.i., while the expression of Id1 gene was at the highest value at 22 day p.i.. Then MyoD gene expression began to gradually decline, at 38 day p.i. returned to normal level. Id1 gene expression decline rapidly after 22 day p.i., returned to normal level at 26 day p.i.. Gene expressions of MyoD and Id1 were both increased after vivo transfection of Ts43 gene with different time, the expression of MyoD gene was increased significantly at 2 day post-transfection (p.t.), while the Id1 gene was at 6 day p.t.. The formation of nurse cells refer to the muscle cells dedifferentiation and re-differentiation. MyoD and Id1 gene ara important regulators of muscle cells. The study found that MyoD, Id1 gene expressions have changed dramatically in cells infected with Trichinella spiralis. MyoD, Id1 gene maybe play important roles for the formation of nurse cells. And MyoD and Id1 gene expression increased in mice skeletal muscle after vivo transfection of Ts43 gene, while further 43ku ES antigen can activate MyoD and Id1 gene expression.In a word, Ts43 gene had a greater relationship with the formation of nurse cells. Ts43 gene was likely to participate the formation of nurse cells by cells regulatory factors. The formation of nurse cells involved a number of ES antigen proteins and cell regulatory factors. To know the real formation mechanism of nurse cells, we should study them in depth.