الملخص
Objective To explore the pattern of early expression and secretion of tissue factor(TF)in vascular endothelial cells induced by heat stress.Methods Thirty SPF-rated C57BL/6 male mice were randomly divided into five groups:the control group and groups of indicated recovery time,including 0,3,6,and 9 h in room temperature after heat stress(n=6).Mice in the heat stress groups were exposed to an animal incubator to reach 42.5℃for core body temperature for heat stroke.We analyzed the histopathological changes in the liver,lung,and kidney tissues with HE staining.We measured the TF mRNA in mice tissues by RT-qPCR and the plasma concentration of TF in mice with a commercial ELISA kit.Human umbilical vein endothelial cells(HUVECs)were placed in a culture incubator to build an in vitro heat stress model.HUVECs were divided into five groups,including a control group and groups of indicated recovery time,including 0,3,6,and 9 h after heat stress.We quantified the expression of TF mRNA and protein in HUVEC cells by RT-qPCR,Western blotting,and immunofluorescence and measured the secreted TF with a commercial ELISA kit.Results No significant pathological injury was observed in the tissues of the control group.Mice treated with heat stress had various degrees of structural injuries and hemorrhagic and inflammatory changes in multiple tissues.Compared to control group,the expression of TF mRNA significantly increased in the kidney of heat stress-treated mice with 0 and 3 h recovery time(1.719±0.018,1.241±0.178 vs.1.000±0.063),the lung with 3 h recovery time(2.444±0.511 vs.1.000±0.106)and the liver with 6 h recovery time(7.312±0.618 vs.1.000±0.147)(P<0.05).The concentration of TF in plasma also sustainedly elevated in mice with 0,3,6,and 9 h recovery time after heat stress as compared to control group[(132.426±17.920)pg/ml,(119.400±10.267)pg/ml,(107.374±13.495)pg/ml,(163.767±22.810)pg/ml vs.(75.479±13.831)pg/ml,respectively,P<0.01].The expression levels of TF mRNA were higher in heat stress HUVECs with 6 h and 9 h recovery time than the control cells(1.905±0.354,2.564±0.297 vs.1.000±0.097,P<0.01).Secreted TF in the supernatant from HUVECs treated with heat stress and different recovery time also increased significantly[(36.309±4.101)pg/ml,(38.425±5.484)pg/ml,(41.655±4.380)pg/ml,(43.586±4.718)pg/ml vs.(14.996±0.254)pg/ml,P<0.01].Conclusion Heat stress increased early expression and secretion of TF in vascular endothelial cells.Vascular endothelial cells may be a main source of circulating TF in heat stroke.
الملخص
Aim To investigate whether Scutellarin ( Seu ) could alleviate LPS + ATP induced inflammation and pvrolysis and the underlying mechanisms.Meth¬ods Primary human umbilical vein endothelial cells (HUVCEs) were pretreated with different concentra¬tions of Scu for 1 h, 2.5 mg • L 1 LPS treatment for 5.5 h, and then 2.5 mmol • L 1 ATP was activated for 0.5 h.The damage rate and activity of endothelial cells were detected by MTT method and determination of LDH content in cell supernatant.ELISA was used to detect the contents of IL-ip,lL-18 and HMGB-1 in su-pernatant.The expression levels of inflammation and pyroptosis related proteins including NLRP3 , ASC , pro- caspase-1 , caspase-1 p20 and GSDMD-NT were detec¬ted by WB method.The expression of pro-caspase-1 , caspase-1 p20 and GSDMD-NT protein and the contents of IL-ip,IL-18 and HMGB-1 in supernatant were de¬tected by Scu and pan caspase receptor inhibitor ( Z-YVAD-FMK).Results Compared with LPS + ATP group, Scu concentration-dependently improved ATP- induced cytotoxic damage, increased cellular activity, reduced the release of 1L-1(3, IL-18, HMGB-1 , down- regulated NLRP3, ASC, pro-caspase-1 , GSDMD-NT protein level to inhibit the activation of pro-caspase-1 , and the effect was comparable to Z-YVAD-FMK.The inhibitory effect of Scu on pro-caspase-1 activation was not significantly different from that of Scu/Z-YVAD- FMK combined group, but significantly inhibited the production of pro-caspase-1.Conclusions Scutellarin can reduce the formation of NLRP3 inflammasome com¬positions and inhibit pro-caspase-1 activation to im¬prove LPS + ATP induced endothelial cell inflammation injury and pyrolysis, which provides a new basis for the clinical prevention and treatment of atherosclerosis.
الملخص
<p><b>OBJECTIVE</b>To compare the differences in the efficacy on distant version of naked eye in the patients of juvenile myopia between rotating manipulation and lifting-thrusting manipulation of acupuncture.</p><p><b>METHODS</b>One hundred and twenty cases (240 eyes) were randomized into a rotating manipulation group and a lifting-thrusting manipulation group, 60 cases (120 eyes) in each group. Additionally, a corrective lenses group, 60 cases (120 eyes), was set up as the control. In both manipulation groups, Cuanzhu (BL 2),Yuyao (EX-HN 4), Sizhukong (TE 23), Taiyang (EX-HN 5), Fengchi (GB 20), Zusanli (ST 36), Guangming (GB 37) and Sanyinjiao (SP 6) were punctured, but stimulated with rotating manipulation and lifting-thrusting manipulation respectively three times per week, 10 times as a treatment session and totally one session was required. In the corrective lenses group, the glasses were applied at daytime. The clinical efficacy and the changes in distant vision of naked eye before and after treatment were compared among the three groups.</p><p><b>RESULTS</b>The total effective rate was 87.5% (105/120) in the rotating manipulation group, which was better than 69.2% (83/120) in the lifting-thrusting manipulation group (P < 0.05). The distant vision of naked eye was improved apparently in the rotating manipulation group and the lifting-thrusting manipulation group after treatment (both P < 0.05). But it was not improved in the corrective lenses group (P > 0.05). The distant vision of naked eye was improved more apparently after treatment in the rotating manipulation group as compared with that in the lifting-thrusting manipulation group (0.75 +/- 0.23 vs 0.68 +/- 0.24, P < 0.05). For 96 cases (192 eyes) with acupuncture treatment, in 3-month follow-up, 87.0% (167/192) of the cases maintained the stable vision as the original level and 13.0% (25/192) of them were reduced in the vision In the acupuncture groups, it was found that the improvement of distant vision of naked eye was more obvious after treatment with younger age, better basic vision and shorter duration of sickness (all P < 0.05).</p><p><b>CONCLUSION</b>Acupuncture achieves the positive and sustainable clinical effect on juvenile myopia, and the results of rotating manipulation are superior to that of lifting-thrusting manipulation. Age, basic vision and duration of sickness impact the clinical efficacy.</p>