Effect On Traditional Chinese Medicine Effective Components Of Porcine Alveolar Macrophage (3D4/2) Function And Its Anti-Porcine Reproductive And Respiratory Syndrome Virus (PRRSV) Mechanism

Objective:Porcine reproductive and respiratory syndrome (PRRS) is a common swine diseases having a strong infectious. It caused by porcine reproductive and respiratory syndrome virus. A typical characteristic of the virus is vulnerable to variation, when outbreak, it will cause huge economic losses to the porcine industry. Currently for the disease mainly take the form of vaccination for prevention, however, there is no effective treatment for the affected individual methods. In this study, Three kinds of natural compound chlorogenic acid, forsythoside A and epigoitrin were extracted from common herbs honeysuckle, forsythia and Radix from having antiviral activity. In 3D4/2 cells in vitro model to study whether the three Chinese high-performance components have a non-specific immunity enhancing lung macrophages. Is able to inhibit the effect of porcine reproductive and respiratory syndrome virus infection on cell, is able to inhibit apoptosis caused by the PRRSV and to explore the antiviral mechanism by using the main component of traditional Chinese medicine.Methods:(1) Identification of 3D4/2 cells by immunohistochemical methods.(2) Determination the maximum safe concentration on the 3D4/2 cells vitro model in Chinese medicine compound by using of CPE method (cytopathologic effect, CPE) and MTT (3-(4,5-dimethyithiazol-2-yl)-2,5-diphenyltetrazolium bromide, MTT) method.(3) Determination of the role of PRRSV infection on the 3D4/2 cells by immunocytochemistry, and Chinese medicine compound on the virus inhibitory effect in 3D4/2 cells on the maximum safe concentration.(4) Determination of Chinese medicine compound on phagocytosis effects by neutral red method. Determination of traditional Chinese medicine compound of macrophages to secrete interleukin 6 (IL-6) interleukin 10 (IL-10) and tumor necrosis factor alpha (TNF-α) expression of the quantity change by ELIS A method.(5) Determination of add Chinese traditional medicine compound of the role of apoptosis induced in PRRSV effect by TUNEL method.Results:Herbal Extracts epigoitrin, forsythoside A and chlorogenic acid have in vitro anti-PRRSV role. The maximum safe concentration of chlorogenic acid:250μg/mL, the maximum safe concentration forsythoside A is:125μg/mL, epigoitrin maximum safe concentration:1000μg/mL. Under the maximum safe concentration adding chlorogenic acid, forsythoside A and epigoitrin on the 3D4/2 cells showing the role of PRRSV. Wherein chlorogenic acid and forsythoside A can effectively enhance the phagocytic capacity of 3D4/2 cells, chlorogenic acid, forsythoside A and epigoitrin able to enhance the secretion of TNF-a and IL-6 while suppressing IL-10 secretion, and chlorogenic acid, forsythoside a and epigoitrin can be suppressed by the PRRSV caused 3D4/2 cells apoptosis.Conclusion:Chinese medicine extraction of natural compounds of chlorogenic acid, forsythoside A and epigoitrin has strong resistance of PRRSV in vitro, and chlorogenic acid, forsythoside A and epigoitrin can strengthen the nonspecific immunity of 3D4/2 cells, chlorogenic acid, forsythoside A and epigoitrin may be strengthen nonspecific immune cells, and can directly inactivated PRRSV and inhibit apoptosis induced by PRRSV.