| Polygonum Cillinerve (Nakai) Ohwi is one of the Chinese herb, it has the function ofheat-clearing and detoxifying, analgesia, hemostasic, and possesses antibacterial and antiviruseffects. It has extensive application on clinic.The Polygonum Ciliinerve (Nakai) Ohwi wasused as materials in this study, the Polygonum Cillinerve polysaccharide was extracted bywater frying alcohol sinking method, the immunological enhancement in chicken in vivo andanti-TGEV activity in vitro were studied for providing theoretical evidences for theexploitation of Polygonum Ciliinerve polysaccharide as immunoenhancer and anti-virus drug.In immune enhancement experiment, taking the chicken which drinking water normally ascontrol, APS group as positive control, and the chicken were divided into five groups: controlgroup, APS group, Polygonum Cillinerve Polysaccharide groups (high, middle and lowdoses). The thymus, spleen and bursa fabricius indexes, histological development state wereobserved, and the serum IgG, LYS content were determined and peripheral blood lymphocyteproliferation test were taken to study the immune enhancement of PCP in chicken. In vitroexperiment, to evaluate the antiviral effect of Polygonum cillinerve polysaccharide (PCP) onTransmissible Gastroenteritis Virus (TGEV), the PCP was pro-seeded, co-seeded orpost-seeded with TGEV in Swine testicular (ST) cells, then the anti-virus adsorption,inactivation and inhibition of the proliferation of PCP were analyzed by MTT and Real-timePCR.The results showed that:(1)Compared with control group, after7d,14d,21d and28d dosing,the middle dosagePCP group and APS group can increase the thymus, spleen, bursa fabricius index of chickenextremely significantly (p<0.01); The high dosage PCP group can increase the thymus,spleenindex of chicken extremely significantly (p<0.01), increase the bursa fabricius index ofchicken significantly (p<0.05); The low dosage PCP group could increase the spleen indexof chicken extremely significantly (p<0.01), and increase thymus and bursa fabricius index ofchicken significantly(p<0.05). Histological morphological observation showed that thechicken in middle dosage PCP group, the thickness of thymus cortex and the density oflymphocytes were the largest, the thymic corpuscle in became big and obvious; Theproportion between splenic white pulp and red pulp was significantly higher than that ofnormal control group, spleen nodules in white pulp were bigger and more, splenic nodules of germinal center became obvious, the lymphoid cell surrounding periarterial lymphatic sheatharranged densely; The volume of lymphoid follicles in bursa of fabricius and its lymphocytedensity were significantly increased.It showed that middle dosage PCP can promote thethymus, spleen, bursa of fabricius growth in chicken. The results of ELISA and MTT showedthat the content of serum IgG, LYS, and peripheral blood lymphocyte proliferation abilitywere significantly improved in the middle dosage PCP group. Experiment results showed that500mg/kg PCP was the best dosage.(2) According to the results of MTT, the maximum cytotoxic concentration (TC0) of PCPin ST cells was20μg/mL, and at the maximum safe concentration, the anti-virus adsorption ofTGEV in vitro was the best, the inhibition rate was85.2%, followed by the inhibition of theproliferation, the inhibition rate was78.4%, finally is the direct inactivation, the inhibition ratewas74.4%, and the inhibition rate was positively correlated with the concentration. Moreover,Real-time PCR detection indicated that compared with the virus control, three administrationmethods of PCP can significantly reduce relative mRNA expression of TGEV at thetranscriptional level（p<0.01），and with dose-dependent characteristic.The results suggested that500mg/kg of PCP has the best immune enhancement inchicken,20μg/mL PCP can inhibit the proliferation of TGEV in ST cell, and the preventioneffect which drug pro-seeded was the best. |
PDF Full Text Request