The Expression Developmental Patterns Of Adiponectin On Immune Organs Of Wannanhua Pigs And Its Effects On Immunological Function

Wannanhua pigs were superordinary local pigs of Anhui province. The production traits and genetic qualities of Wannanhua pigs were special in Chinese local pigs. Adiponectin (Adp) was a sole idio-protein of adipokines family, it not only involved in regulating glucose and metabolism, inhibition of inflammation through variety ways, but also acted on almost all organs and cells, especially immune cells, involed in body immune response. Spleen(SP) as the body`s largest peripheral lymphoid organ, involed in body cell immunity and humoral immunity. SP is a sites of lymphocytes and other immune cell generation, differentiation, maturation, settlement, proliferation and immune reponse. As the most important immune organ of the body, cervical lymph nodes (CLN) were the sites to accept the immune response antigen stimulation, with filtration, proliferation, and immune function. In this study, the relation of developmental patterns of blood immune parameters and Adp mRNA expression in SP and CLN was investigated. And the inner relationship of Adp and immune system in Wannanhua pigs was approached. The effects of recombinate Adp(rAdp) on cultured spleen cells of pigs were also studied in this paper in order to explore the influence of Adp on immune system in Wannanhua pig, and its possible mechanism.Five Wannanhua boars and five Wannanhua gilts were sampled at0,30,45(weaning),90, and180days of age respectively. The test animals were procured from Wannanhua pigs breed-conservation base in Yixian county, Anhui province. Pigs were allowed an access to spontaneous feed and water. Pigs were weighed and slaughtered after12hours fasting(except0d). Blood samples were collected via heart. The spleen and Lymph nodes were weighed and taken immediately to be frozen in liquid nitrogen and then stored at-80℃. The level of blood TNF-α and IL-6were determinated by enzyme-linked immunosorbent assay(ELISA). The level of blood immunoglobulins were determinated by turbidimetric immunoassay. The abundances of Adp, adiponectin receptor1(AdpR1), and adiponectin receptor2（AdpR2）, TNF-α and IL-6mRNA in SP and CLN, and TNF-α, IL-6mRNA in subcutaneous(SC) and perirenal(PR) adipose tissue were detected by real-time fluorescence quantitative RT-PCR. The results were as follows:1Developmental patterns of blood immune parameters in Wannanhua pigsSerum TNF-α and IL-6of Wannanhua pigs were changed with age significantly (P<0.01), there were sexual dimorphism after45d (P <0.05or P <0.01). IgG in serum of Wannanhua pigs were changed with age significantly(P<0.01), but IgA and IgM in serum showed no developmental change in general. Serum IgG, IgA and IgM was higher in Wannanhua gilts than those in Wannanhua boars(P <0.05or P <0.01).2Developmental patterns of Adp, AdpR mRNA and immune related genes expressions in Wannanhua pigs and their tissue specificity and sexual dimorphism.(1) Adp mRNA expression in SP changed with age significantly(P<0.05), which were significantly lower in SP than those in CLN (P <0.05) AdpR1and AdpR2mRNA expression in SP and CLN were changed with age significantly(P<0.01), and AdpR2mRNA expression were significantly higher in CLN than those in SP(P <0.05).(2) TNF-α mRNA expression in CLN and SC adipose tissue changed with age significantly(P <0.05or P <0.01), which were significantly higher in SP than those in CLN (P <0.05) before ablactation, but then opposite after weaning. TNF-α mRNA expression in SC adipose tissue was significantly higher than that in PR adipose tissue in general IL-6mRNA expression in SP, CLN, SC adipose tissue and PR adipose tissue all changed with age significantly(P <0.05or P <0.01), which were significantly higher in SP than those in CLN(P <0.05).(3) Although the sexual dimorphism were found in apart genes or apart days of age, Adp、AdpR1、AdpR2、 TNF-α and IL-6mRNA expression both in SP and CLN had no significant differences between Wannanhua gilts and boars in general. TNF-α and IL-6mRNA expression in SC adipose tissue and PR adipose tissue had the same situation.(4) Significant positive correlation was found between Adp and AdipoR1in SP (r=0.39, P<0.05）, and significant positive correlation were found between TNF-α and IL-6in SP、 CLN and SC adipose tissue(r=0.54, P<0.01r=0.70，P<0.01r=0.64, P<0.01).The results suggested that blood biochemical parameters of Wannanhua pigs show special developmental patterns and sexual dimorphism to some extent. Adp、A dpR1、A dpR2、T NF-α and IL-6may interact and regulate organism immune function together.3Effect of adiponectin on cultured spleen cell of Wannanhua pigs and its possible mechanismSpleen cells were isolated from spleen tissue at10day s of age of Wannanhua pig. Then cells were cultured for2days, MTT method were used to detect viability of cells. When cells were cultured for5days, the cells were treated with1μg/ml5μg/ml10μg/ml and15μg/ml recombinate adiponectin(rAdp) for12h (acute treatment) or24h (chronic treatment) respectively, and MTT method were used to detect viability of cells. The cells were treated with rAdp, rAdp+NAC and NAC for12h or24h, then ELISA method were used to detect the level of TNF-α and IL-6in cell supernatant fluid. The viability of cells was decreased in all experimental groups, and the level of TNF-α and IL-6had a decreasing trend in rAdp treatment group, but had a rising trend in rAdp+NAC treatment group, in which, the level of IL-6in Adp+NAC treatment group was significantly higher than Adp treatment.The results indicated that recombinate adiponectin may depress the viability of spleen cell and inhibit TNF-α and IL-6secretion of spleen cell.