Effects Of Stems And Leaves Of Panax Notoginseng On The Expression Level Of Toll-like Receptors In The Liver Of Duzang Pigs

The shortage of feed resources restricts the high-quality development of animal husbandry.In order to alleviate the shortage of feed resources,the development of unconventional feed resources is imminent.As a potential unconventional feed resource,the stems and leaves of Panax notoginseng have great development and utilization value.Therefore,30 Duzang pigs(Duroc♂×Diqing Tibetan pig♀)were selected in this study,and were randomly divided into 3 groups with 10 pigs in each group.The control group was fed the basal ration,and the experimental group I and II were fed the experimental ration added with 10% and 20% Stems and Leaves of Panax notoginseng in the basal ration,respectively.After 32 days of feeding,5 heads were randomly selected for slaughter in each group.The liver was collected,and the m RNA expression of nine members of the TLR gene family and their key signaling pathway proteins and downstream cytokines were detected by real-time fluorescent quantitative PCR method,and then the correlation analysis was performed on the expression levels of each gene to explore the effects stems and leaves of Panax notoginseng Effects on gene expression of TLRs signaling pathway in Duzang pigs.The main results are as follows:(1)Effects of stems and leaves of Panax notoginseng on the growth performance of Duzang pigs: The stems and leaves of Panax notoginseng had no significant effect on the average feed intake,average daily gain and feed-to-weight ratio of Duzang pigs(p>0.05).(2)SMART program protein domain prediction shows: Nine genes of porcine TLRs(TLR1-9)all contain TIR domains,among which TLR1/2/3/4/5/6/7/8 contain LRRs motif,LRR-CT motif,transmembrane domain and TIR structure domain,TLR9 lacks the LRR-CT domain and the transmembrane domain,but the number of LRRs varies significantly.(3)Effects of stems and leaves of Panax notoginseng on the expression of genes related to TLR signaling pathway in Duzang pig liver: The expression level of TLR1 gene m RNA in test group I was significantly higher than that in test group II(p<0.05).The expression level of TLR3 gene m RNA in the experimental group Ⅰ was significantly higher than that in the control group(p<0.05).The m RNA expression levels of TLR4,TLR7,My D88,TRIF,IFN-α and IL-10 genes in the test group I were significantly higher than those in the control group and the test group II(p<0.05).The m RNA expression levels of TLR5,TLR9,IL-1β and TNF-α genes in the experimental group Ⅰwere significantly lower than those in the control group(p<0.05),and the m RNA expressions of TLR5,TLR9 and TNF-α genes in the experimental group Ⅱ were significantly lower than those in the control group(p<0.05).The m RNA expression of TLR6 and IFN-β genes in the test group II was significantly higher than that in the control group and the test group I(p<0.05),and the IFN-β gene m RNA expression in the test group I was significantly higher than that in the control group(p<0.05).The m RNA expression of TLR8 gene in the experimental group Ⅰ and experimental group Ⅱwas significantly higher than that in the control group(p<0.05).(4)Gene correlation analysis of TLR signaling pathway in Duzang pig liver: TLR1,TLR9 and My D88 were significantly positively correlated(p<0.05),and TLR6 gene was extremely significantly negatively correlated with My D88 gene(p<0.05).TLR4,TLR7,TLR8 were significantly positively correlated with TRIF(p<0.05),TLR5 gene was significantly negatively correlated with TRIF gene(p<0.05).TLR7,TLR8 and IL-10 were extremely significantly and strongly positively correlated(p<0.01),and TLR5 gene was significantly negatively correlated with IL-10 gene(p<0.05).TLR5,TLR9 were positively correlated with TNF-α(p<0.05),and TLR6 gene was significantly negatively correlated with TNF-α gene(p<0.01).TLR4,TLR7,TLR8 genes were significantly positively correlated with IFN-α gene(p<0.05).TLR8,TLR6 were positively correlated with IFN-β(p<0.05),and TLR5,TLR9 genes were extremely significantly negatively correlated with IFN-β(p<0.01).In summary,adding 10% and 20% of Panax notoginseng stems and leaves in the diet did not significantly affect the growth performance of Duzang pigs,but could promote the expression of TLR1,TLR3,TLR4,TLR6,TLR7 and TLR8 in the liver of Duzang pigs.The expressions of key downstream proteins My D88,TRIF,and downstream cytokines type I interferon and IL-10 in the signaling pathway,while IL-1β and TNF-α decreased significantly,contributing to the improvement of immunity.