ABSTRACT
OBJECTIVE To study the effects of Wenyang huazhuo tongluo formula conta ined serum on the expression and methylation of retinoic acid related nuclear orphan receptor （RORγt），and to explore the mechanism of its regulation of Th 17 cell proliferation in the treatment of scleroderma. METHODS Wistar rats were given 15，30，60 g/（kg·d）Wenyang huazhuo tongluo formula intragastrically to prepare different doses of drug-contained serum ，and peripheral blood of scleroderma patients were collected to sort Th 17 cells. Using blank serum as blank control ，methyltransferase inhibitor decitabine （DCA）as positive control ， Th17 cells were treated with different concentrations of drug-contained serum ，and then the proliferation of Th 17 cells was detected by CCK- 8；mRNA expressions of RORγt and IL-17 were detected by qRT-PCR ，and the protein expression of RORγt was detected by Western blot assay ；the protein expression of IL- 17 in the cell supernatant was detected by ELISA ，and the methylation level of RORγt gene promoter was detected by methylation-specific PCR. The transcriptional activity of RORγt gene promoter was detected by dual-luciferase assay. RESULTS Compared with blank control ，Wenyang huazhuo tongluo formula contained serum and DCA could inhibit the proliferation of Th 17 cells（P＜0.05），reduced the mRNA and protein expressions of RORγt and IL-17，enhanced the methylation level of RORγt gene promoter and attenuated ， the transcription activity of RORγt gene promoter. Except for mRNA expression of Th 17 and the methylation level of RORγt gene promoter in drug-contained serum low-dose group ，there were statistical significance in above indexes of other groups（P＜0.05）. CONCLUSIONS Wenyang huazhuo tongluo formula can promote the methylation lev el of RORγt gene，and inhibit the expression of RORγt and the secretion of IL-17，so as to inhibit the proliferation of Th 17 cells.
ABSTRACT
OBJECTIVE：To investigate the protective effects of drug-contained serum of Xiaoxuming decoction （XXM）on astrocyte of oxygen and glucose deprivation model rats ，and to explore its mechanisms. METHODS ：The astrocytes of rats were randomly divided into control group ，model group and XXM low-dose ，middle-dose，high-dose groups. The cells in the control group were not treated ；after 2.5 h of OGD ，model group and XXM low-dose ，middle-dose，high-dose groups were reoxygenated for 0，3，6，12 h in 0（i.e. the model group was not added with drugs ），2.5％，5％，10％ of XXM ，respectively. The content of lactate dehydrogenase （LDH）was detected by colorimetry. The reactive oxygen species （ROS）level was detected by fluorescence probe method ，and the expression of Manganese superoxide dismutase （MnSOD）was determined by immunofluorescence double staining method in control group ，model group and XXM high-dose group after 12 h of reoxygenation following OGD. RESULTS ： The content of LDH in the control group was always kept at a low level ；LDH content in the model group gradually increased from （110.99±17.06）U/L to （436.64±55.29）U/L after 0-12 h of reoxygenation following OGD ，which was significantly higher than that in the control group （P＜0.05）. Compared with model group at the same time point after reoxygenation following OGD ，the contents of LDH in the cells of XXM low-dose ，medium-dose and high-dose groups were decreased to different extents ，and showed a time-and dose-dependent trend. The contents of LDH in XXM groups at 6 and 12 h after reoxygenation following OGD were significantly lower than that of the model group （P＜0.05）. At 12 h after reoxygenation following OGD ，the levels of ROS in model group were significantly higher than control group ， while the level of MnSOD was significantly lower than control group（P＜0.05）. The level of ROS in XXM high-dose group hospital.sh.cn was significantly lower than model group ，while the level of MnSOD was significantly higher than model group （P＜0.05）.. CONCLUSIONS：XXM can protect astrocyte by up-regulating sh.cn levels of MnSOD ，scavenging excessive oxygen free radicals ， to relieve the OGD induced astrocytic injury ，with protective effect.
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Objective To study the effects of serum containing Psoralea on proliferation and melanogenesis in A375 human melanoma cells by serum pharmacological method. Method A375 human melanoma cells were cultured in vitro. Different concentrations of drug-contained serum were added to the culture medium during logarithmic growth phase. Melanocyte culture was determined by MTT and NaOH method. Results There was a significant difference between the control and the drug-contained serum test group on proliferation, and there was significant increase in 20% serum drug-contained serum on melanogenesis (P
ABSTRACT
Objective To study the effect of Qingfei oral liquid contained serum on the transforming growth factor-?1 (TGF-?1) and platelet-derived growth factor-BB (PDGF-BB) mRNA gene expression of human embryonic lung fibroblast cells infected by adenovirus (ADV) type 3I and 7b. Method TGF-?1 and PDGF-BB mRNA expression of human embryonic lung fibroblast cells infected by ADV type 3I and 7b were determined by in situ hybridization before and after treated with Qingfei oral liquid contained serum. Results ADV could up-regulate TGF-?1 and PDGF-BB mRNA of human embryonic lung fibroblast cells (P