Study on antipyretic effect of rhubarb on rats and its antipyretic ingredients / 中国中药杂志

A combination of LC-MS technology and activity evaluation was used to identify the antipyretic ingredients in rhubarb. The rat model of fever was established with dried yeast and then was administered ethanol extract and different polar fractions of rhubarb. Next, the anal temperature of these rats was measured and recorded at 0.5, 1, 2, 3, 4 and 5 h after administration, and the inhibition rate of each part on the rise of body temperature was calculated. The inhibition rate is higher and the antipyretic effect is better. The chemical composition of the effective fraction was analyzed with UPLC-ESI-Orbitrap-MS/MS technology. Compared with the model group, the increase of body temperature of ethanol extract group all reduced at each measurement time especially after 3 h, and the inhibition rate were 38.7%(P<0.05), 78.2%(P<0.01) and 72.4%(P<0.01) at 3 h, 4 h, and 5 h after administration, respectively. Both n-butanol and water fraction showed some antipyretic activity in the early stage, with the inhibition rate of 28.1%(P<0.01) and 24.9%(P<0.05) at 1 h after administration, respectively, while other fractions were not active. Thirty-three and twelve compounds were identified from n-butanol and water fraction by LC-MS/MS analysis, respectively, including ten tannins, fifteen anthraquinone glycosides, four anthrone glycosides, one phenolic glycoside, one naphthaline derivative, one anthraquinone and one sucrose. These results revealed that rhubarb had antipyretic activity on rats, and tannin and anthraquinone glycosides were the main active ingredients inside.

Protective effect of Sestrin2 on apoptosis of dendritic cells induced by high mobility group box-1 protein / 解放军医学杂志

Objective To investigate the potential role of Sestrin2 (SESN2) in regulating the apoptosis of dendritic cells (DCs) induced by high mobility group box-1 protein (HMGB1).Methods DCs (the murine DC cell line DC2.4) were cultured with or without HMGB1 stimulation (cultured with 10ng/ml HMGB1 for 8,24 and 48 hours,or cultured with HMGB1 for 48 hours at different concentrations of 1,10 and 100ng/ml,respectively,n=4).The protein level of SESN2,cleaved-caspase-3 and Bcl-2 were analyzed with Western blotting.Localization of SESN2 in cells was observed under confocal laser microscope (LSCM).Cell apoptosis was analyzed with flow cytometry.In addition,DC2.4 cells were transfected with lentivirus containing SESN2 LV-RNA,SESN2 siRNA sequence expressing plasmids or blank vector (NC,NS,n=4).These cells were then stimulated with HMGB1 (100ng/ml)for 48 hours,and the apoptosis was accessed as mentioned above.Results Compared with the control group,the expression of SESN2 was obviously up-regulated after HMGB1 (10ng/ml) stimulation for 24 and 48 hours (P＜0.05).In a dose-dependent response,the expression of SESN2 was markedly enhanced in treatment with 1,10,100ng/ml HMGB1 for 48 hours (P＜0.05).Compared with the control group (7.35％ ± 1.33％),the percentage of apoptosis was significantly increased with 10,100ng/ml HMGB1 for 48 hours [(17.02％ ± 4.85％,17.48％ ± 4.04％,respectively,P＜0.05 or P＜0.01].After transfection,compared with blank vector group,the apoptosis of SESN2 siRNA group obviously elevated [(65.96％ ± 2.50％) vs.(50.01％ ± 2.07％),P＜0.05],and cleaved-caspase-3 expression significantly increased while Bcl-2 expression obviously decreased.In SESN2 LV-RNA group,the apoptosis significantly decreased [(35.57％ ± 1.69％) vs.(49.04％ ± 4.87％),P＜0.05],and cleaved-caspase-3 expression decreased and Bcl-2 expression obviously increased compared with blank vector group (P＜0.05).Conclusion SESN2 has a protective effect against HMGB 1 induced apoptosis of D C2.4 cells.

Expression of Beclin1 and HLA Ⅰ , Ⅱ in human SKOV3 ovarian cancer cells and their correlation / 临床与实验病理学杂志

Purpose The aim of this study was to investigate the relationship between autophagy gene Beclin1 and immune response effector classical HLA Ⅰ,Ⅱ in SKOV3 cells.To explore the role of Beclin1 in immunity in ovarian cancer cells which were transfected with the vector of Beclin1.Methods RT-PCR and Western blot were used to detect the expression of Beclin1 and HLA Ⅰ,Ⅱ in SKOV3 cells.Fluorescence microscope was carried out to observe the unique autophagosome in SKOV3 cells.MTT was used to analyze the proliferation of the Beclin1 over-expressed SKOV3 cells.Results Transfection SKOV3 cells with Beclin1 vector could induce Beclin1 transcription and translation approximately 5 and 2 times compared with empty vector group respectively.The autophagosome stained by MDC was observed by fluorescence microscope.And much more green fluorescence signal was observed in Beclin1 vector group.RT-PCR and Western blot indicated that HLA Ⅰ,Ⅱ induced by transfection with extrinsic Beclin1.The allelic transcriptions of HLA Ⅰ-A,B,C and HLA Ⅱ-DP,DQ,DR in extrinsic Beclin1 group were approximately 2,1.6,3 and 2,6,3 times compared with empty vetcor group or untreated group,respectively.The results of Western blot showed that HLA Ⅰ,Ⅱ in Beclin1 vector group induced as much as 2 and 1.6 times compared with empty vetcor group or untreated group,respectively.The results of MTT showed that the proliferation of SKOV3 cells treated with Beclin1 vector was significantly suppressed.The percentage of suppression in Beclin1 vector group at 24 h,48 h,72 h and 96 his42.6％,37.8％,24.35％,14.81％ compared with untreated group or empty vector group respectively.Conclusion The enhancement of autophagy by over-expression of Beclin1 could induce HLA Ⅰ,Ⅱ transcription and translation in SKOV3 cells.The expression of HLA Ⅰ,Ⅱ may be responsible for triggering the immune response in ovarian cancer.Over-expression of Beclin1 could inhibit the proliferation of SKOV3 cells which were transfected with extrinsic Beclin1.