不同剂量LPSp联合HBsAg对孕鼠IL-4、IL-6、IFN-γ及TLR4的影响

发布时间：2018-03-03 17:00

本文选题：LPSp　切入点：IL-4　出处：《广西医科大学》2012年硕士论文　论文类型：学位论文

【摘要】：目的：研究革兰阴性非致病成团泛菌脂多糖(LPSp)联合乙肝表面抗原(HBsAg)对孕鼠血清IL-4、IL-6及IFN-γ的影响,并探讨其对孕鼠胎盘Toll样受体4 (TLR4)表达的影响。方法：将50只孕鼠随机分为5组：对照组、HBsAg组、联合低剂量组、联合中剂量组、联合高剂量组,每组10只。于妊娠的第1天和第14天,对照组肌注生理盐水100ul、HBsAg组肌注HBsAg 100ul、联合低剂量组肌注HBsAg 100ul+LPSp 5ug、联合中剂量组肌注HBsAg 100ul+LPSp 15ug、联合高剂量组肌注HBsAg 100ul+LPSp 25ug,妊娠第20天时收集孕鼠血清及胎盘。用ELISA方法检测孕鼠血清中IL-4、IL-6及IFN-γ的含量,用免疫组化方法检测孕鼠胎盘中TLR4蛋白的表达,用逆转录-聚合酶链反应(RT-PCR)法检测孕鼠胎盘中TLR4 mRNA的表达。测量新生小鼠身长、体重及脑重。结果：(1)联合各剂量组IFN-γ水平均明显高于对照组及HBsAg组,且呈浓度依赖性,P均0.01；联合各剂量组IL-4、IL-6水平均明显高于对照组,P均0.01,但联合高剂量组IL-4、IL-6水平与HBsAg组比较差异无统计学意义,P均0.05；HBsAg组IFN-γ水平明显低于对照组,IL-4、IL-6水平高于对照组,P均0.01。(2)LPSp刺激后TLR4蛋白表达水平明显增高,联合各剂量组TLR4蛋白表达水平明显高于对照组及HBsAg组,P均0.01。(3)LPSp刺激后TLR4mRNA表达水平明显增高,联合各剂量组TLR4mRNA表达水平明显高于对照组及HBsAg组,P均0.01。(4)各组胎鼠的身长、体重、脑重等生长发育指标比较差异均无统计学意义,P0.05。结论：(1)LPSp对胎鼠的生长发育无明显不良影响,具有一定的安全性。 (2) LPSp可调节孕鼠血清中Thl/Th2细胞因子水平,且呈剂量依赖性,高剂量LPSp可使细胞因子IFN-γ反应增强,而低剂量LPSp可使细胞因子IL-4、IL-6反应增强。 (3)LPSp能增强TLR4蛋白和TLR4mRNA在孕鼠胎盘中的表达,而LPSp可能不会造成胎盘形态学的改变。
[Abstract]:Aim: to study the effect of lipopolysaccharide (LPSp) combined with hepatitis B surface antigen HBsAg (HBsAg) on serum IL-4- 6 and IFN- 纬 in pregnant rats, and to investigate its effect on the expression of Toll like receptor 4 (TLR4) in placenta of pregnant rats. Methods: fifty pregnant rats were randomly divided into five groups: control group, low dose group, combined medium dose group, combined high dose group, 10 rats in each group. The control group was intramuscularly injected with normal saline 100ul LPSp, the low dose group was intramuscularly injected with HBsAg 100ul LPSp 5ug, the middle dose group was intramuscularly injected with HBsAg 100ul LPSp 15ugand the high dose group was injected with HBsAg 100ul LPSp 25ug. the pregnant rat serum and placenta were collected on the 20th day of gestation. ELISA prescription was used. Methods to detect the levels of IL-4, IL-6 and IFN- 纬 in serum of pregnant rats. The expression of TLR4 protein in placenta of pregnant mice was detected by immunohistochemical method, the expression of TLR4 mRNA in placenta of pregnant rats was detected by reverse transcription-polymerase chain reaction (RT-PCR) method, and the body length, body weight and brain weight of newborn mice were measured. Results the levels of IFN- 纬 in each dose group were significantly higher than those in control group and HBsAg group. The levels of IL-4 and IL-6 in each dose group were significantly higher than those in the control group (P 0.01), but there was no significant difference between the high dose group and the HBsAg group. The level of IFN- 纬 in the combined group was significantly lower than that in the control group. Compared with the control group, the expression of TLR4 protein increased significantly after stimulation with LPSp. The expression of TLR4 protein in each dose group was significantly higher than that in the control group and HBsAg group (P 0.01.3LPSp). The expression of TLR4mRNA in each dose group was significantly higher than that in the control group and HBsAg group (0.01.44)) the body length and body weight of the fetal rats in each dose group were significantly higher than those in the control group and the HBsAg group. There was no significant difference in growth and development indexes of brain weight (P 0.05). Conclusion there is no significant adverse effect on the growth and development of fetal mice by LPSp, and it is safe to some extent. (2) LPSp could regulate the level of Thl/Th2 cytokines in serum of pregnant rats in a dose-dependent manner. High dose LPSp could increase the response of cytokines IFN- 纬, while low dose LPSp could increase the response of cytokines IL-4 and IL-6. LPSp can enhance the expression of TLR4 protein and TLR4mRNA in placenta of pregnant rats, but LPSp may not cause the change of placenta morphology.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R392.1

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