| Lipopolysaccharide (LPS) is a component of the cell wall of Gram-negative bacteria, a common TLR4 and TLR2 ligands. it is also a typical representative of the pathogen associated molecular patterns (PAMPs), which can induce the same immune stress as Gram-negative bacteria infection. In this study, female rabbits were injected with saline (LPS carrier) or LPS instead Gram-negative bacteria infection under natural condition. The aim of this study was to reveal the effect of LPS on expression level of TLR4,TLR2, and downstream immune cytokines, IL-1β, IL-6 and TNF-α in hypothalamus, oviduct and ovary.The expression level of TLR4, TLR2, IL-1β, IL-6 and TNF-α were analyzed by real-time PCR and the concentration of IL-1β, IL-6 and TNF-α were examined by ELISA.Real-time PCR results showed:TLR4, TLR2, IL-1β, IL-6 and TNF-α were expressed in hypothalamus, oviduct and ovary. After injection of saline, TLR4 transcript level at 1.5,3,6 and 12h had no significant difference with Oh(P>0.05), TLR2 in hypothalamus 6h and oviduct 3h was significantly higher than Oh (P<0.05), the level of transcription of immune cytokines had no significant change compared with the bank group in each organization after injection 1.5-12h(P>0.05); After LPS inducted, TLR4 transcript level was significantly increased only in ovary compared with the control group (P<0.05), but in the oviduct after injection 1.5h,3h and 12h was significantly lower than Oh (P<0.05), it was significantly higher than Oh that TLR2 transcript level in hypothalamus 3h and ovaries 12h (P<0.05), the transcription level of IL-1β in the hypothalamus 1.5h,3h and 12h, ovary 1.5-12h and oviduct 6h was significantly higher than Oh (P<0.05), the transcription level of IL-6 in hypothalamus 1.5～12h, ovary and oviduct 1.5-3h was significantly higher than Oh (P<0.05), and the transcription level of TNF-α in hypothalamus 1.5h,3h and 12h, ovary 1.5h and oviduct 1.5-3h was significantly higher than oh (P<0.05); Experimental group compared with the control group showed that, TLR4 transcript levels only in the ovary 12h was significantly higher (P<0.05), TLR2 in hypothalamus 3h was significantly higher (P<0.05), but 6h was significantly lower than the control group (P<0.05), in ovary 12h was significantly higher (P<0.05), in oviduct 3h was significantly lower than the control group (P<0.05), IL-1β transcript level in hypothalamus 1.5h,3h and 12h, ovary 3～12h, oviduct 6h was significantly higher (P<0.05), IL-6 transcript level in hypothalamus 1.5～12h, ovary and oviduct 1.5～3h was significantly higher (P<0.05), and after injection TNF-α transcript level in hypothalamus 1.5h,3h and 12h, ovary 1.5h and oviduct 1.5-3h was significantly higher than the control group (P<0.05).ELISA detection of cytokine proteins showed:The protein of TLR4, TLR2. IL-1β, IL-6 and TNF-α were expressed in all experimental tissue samples. In the control group, after injection of saline IL-1β and IL-6 in various tissues 1.5～12h, there was no change compared with Oh (P>0.05), TNF-α in oviduct 3-12h was significantly lower than Oh (P<0.05); After LPS injection, IL-1β in hypothalamus 1.5～12h, ovary 1.5～6h, oviduct 1.5h and 6h was significantly higher than Oh (P<0.05), IL-6 in ovary 3h and oviduct 6h was significantly higher than oh (P<0.05), TNF-a in hypothalamus 3～6h and oviduct 1.5～12h was significantly lower than oh (P<0.05); Experimental group compared with the control group showed that, IL-1β in hypothalamus 1.5～12h, ovary 3-6h, oviduct 1.5h,6h was significantly lower than the control group (P<0.05), IL-6 in hypothalamus 12h, ovary 3h, oviduct 6h was significantly higher than the control group (P<0.05), TNF-a in oviduct 1.5h was significantly lower than the control group(P<0.05).Generally speaking, there were TLR4 and TLR2 expression in oviduct, ovary and hypothalamus of rabbits, but after LPS induction,TLR4 transcript levels increased significantly only in the ovary (P<0.05), TLR2 transcript levels increased significantly in both ovary and hypothalamus(P<0.05); And LPS significantly induced the expression level of IL-1β,IL-6 and TNF-α in the hypothalamus, ovary and oviduct (P<0.05), but the immunosuppressant of LPS had obviously different in three organs.Namely, immunoresponse in the hypothalamus response to LPS was more sensitive and the duration of immunoresponse was longer than in the ovary and oviduct. |
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