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Objective To investigate the effect ofDihuangyinzi (DHYZ)-medicated serum on receptor for advanced glycation end product (RAGE)/p38 miotgen-activated protein kinase (MAPK) /nuclear factor (NF)-κB pathway in SH-SY5Y cells induced by Aβ1-42.Methods Male SD rats were randomly divided into normal control group and experimental group (n=20);natural sera medium and DHYZ sera medium were prepared.(1) SH-SY5Y cells were divided into control group,model group and DHYZ treatment group;natural sera medium,natural sera medium+Aβ1-42 oligomer,and DHYZ sera medium+Aβ1-42 oligomer were given to the cells,respectively.Westem blotting was used to detect the protein expressions of NF-κB p65,p38 and phosphorylate (p)-p38.(2) SH-SY5Y cells were given DHYZ sera medium+Aβ1-42 oligomer treatment,and at different time points of Aβ1-42 oligomer treatment (15 min,30 min,60 min,12 h,24 h,48 h and 72 h),Western blotting was used to detect the protein expressions of p38 and p-p38.(3) SH-SY5Y cells were divided into 6 groups:mock-transfected RAGE blank group,transfected RAGE blank group,mock-transfected RAGE model group,transfected RAGE model group,mock-transfected RAGE herb group and transfected RAGE herb group;herb groups were given DHYZ-medicated serum;inflammatory factors,interleukin (IL)-1β,IL-6,and tumor necrosis factor (TNF)-a,were measured by ELISA and cytometric bead array.Results (1) As compared with model group,DHYZ treatment group had significantly decreased NF-κB p65 and p-p38/p38 protein expression.(2) The p-p38 protein expression began to increase 30 min after Aβ1-42 treatment,reached to its peak level 24 h after Aβ1-42 treatment,and began to decrease 48 h after Aβ1-42 treatment.(3) The IL-1β,IL-6 and TNF-α levels were increased significantly in the transfected RAGE model group as compared with those in the mock-transfected RAGE model group (P＜0.05);the IL-1β,IL-6 and TNF-α levels were increased significantly in the transfected RAGE herb group as compared with those in the mock-transfected RAGE herb group (P＜0.05);the IL-1β,IL-6 and TNF-α levels were decreased significantly in the mock-transfected RAGE herb group as compared with those in the mock-transfected RAGE model group (P＜0.05);the IL-1β,IL-6 and TNF-α levels were decreased significantly in the transfected RAGE herb group as compared with those in the transfected RAGE model group (P＜0.05).Conclusion DHYZ-medicated serum could inhibit the RAGE-p38 pathway and improve the inflammatory reaction in Aβ1-42-induced SH-SY5Ycells transfected with RAGE gene to protect the SH-SY5Y cells.
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Objective To investigate the effects of Dihuangyinzi(DHYZ) on behaviors and RAGE/p38 pathway in APP/PS1 mice.MethodTwenty APP/PS1 dementia mice were randomly divided into model group(n=10) and Chinese medicine group(n=10).The blank group was C57 BL/6 J normal mouse(n=10).The mice in Chinese medicine group were intragastric administration with DHYZ (9.75 g·kg-1·d-1).The mice in model group and blank group were treated with distilled water.After 30 days,the abilities of learning and memory of mice were detected by Morris water maze.The expression of amyloid-beta1-42(Aβ1-42) in the hippocampus and cortex was detected by immunohistochemistry.Reactive oxygen species of brain tissue were detected by DCFH-DA Methods in the brain of APP/PS1 mice.Gene expression level of receptor for advanced glycation end products(RAGE) was measured by real-time polymerase chain reaction (RT-PCR) in the cortex and hippocampus of APP/PS1 mice.The expression of phospho-mitogen-activated protein kinases (p38) was analyzed with Western blot and immunofluorescence analysis in the cortex and hippocampus of APP/PS1 mice.Results Behavioral Results showed that DHYZ significantly increased the distance((23.088±7.083)cm) and residence time((1.961±1.230)s)of effective area in Morris water maze on the fifth day(P<0.05,P<0.01)and remarkably increased the number of effective area crossings((1.607±0.405) times) and plats((0.893±0.283) times) in Morris water maze on the fifth day(P<0.01,P<0.05).DHYZ also significantly reduced the intracelluar ROS level(122.611±7.630) in the brain(P<0.01),and DHYZ could depress the expression of RAGE(1.467±0.081,7.983±0.136) and phosphorylation of p38 (0.376±0.026,0.538±0.016)in the cortex and hippocampus of APP/PS1 mice(P<0.01,P<0.05).Conclusions The Results demonstrate that DHYZ can partly improve memory impairment of APP/PS1 mice by the inhibition of RAGE/p38 pathway.
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BACKGROUND: Lychas mucronatus is one scorpion species widely distributed in Southeast Asia and southern China. Anything is hardly known about its venom components, despite the fact that it can often cause human accidents. In this work, we performed a venomous gland transcriptome analysis by constructing and screening the venom gland cDNA library of the scorpion Lychas mucronatus from Yunnan province and compared it with the previous results of Hainan-sourced Lychas mucronatus. RESULTS: A total of sixteen known types of venom peptides and proteins are obtained from the venom gland cDNA library of Yunnan-sourced Lychas mucronatus, which greatly increase the number of currently reported scorpion venom peptides. Interestingly, we also identified nineteen atypical types of venom molecules seldom reported in scorpion species. Surprisingly, the comparative transcriptome analysis of Yunnan-sourced Lychas mucronatus and Hainan-sourced Lychas mucronatus indicated that enormous diversity and vastly abundant difference could be found in venom peptides and proteins between populations of the scorpion Lychas mucronatus from different geographical regions. CONCLUSIONS: This work characterizes a large number of venom molecules never identified in scorpion species. This result provides a comparative analysis of venom transcriptomes of the scorpion Lychas mucronatus from different geographical regions, which thoroughly reveals the fact that the venom peptides and proteins of the same scorpion species from different geographical regions are highly diversified and scorpion evolves to adapt a new environment by altering the primary structure and abundance of venom peptides and proteins.