ABSTRACT
Objective To explore the therapeutic effect of epinephrine combined with acupuncture on anaphylactic shock and its mechanism. Methods Sixty male Kunming mice were randomly divided into normal saline (NS) group, anaphylactic shock model group, and integrated traditional Chinese and Western medicine treatment group with 20 mice in each group. The anaphylactic shock model was reproduced by egg albumin infusion: intraperitoneal injection of 0.25 mL egg albumin (0.01 mmol/L), repeated injection 1 week later, and intravenous injection of 0.5 mL egg albumin through caudal vein on the 3rd week to induce anaphylactic shock. The mice in the NS group were injected with NS. The mice in the treatment group were immediately subcutaneously injected with 0.2 μg of 0.1% epinephrine, and intraperitoneally injected with aminophylline 0.2 mg, combined with acupuncture at Shuigou, Neiguan and Hegu points. Number of died mice in each group were observed at 1, 6, and 12 hours after model reproduction. The mice were sacrificed at 12 hours, the blood was harvested, and the serum tryptase, immunoglobulin E (IgE), tumor necrosis factor-α (TNF-α), interleukins (IL-1 and IL-6) were determined by enzyme linked immunosorbent assay (ELISA). The lung tissues were harvested, and the protein expressions of p65, phosphorylation of p65 (p-p65), and phosphorylation of nuclear factor-κB inhibitor α (p-IκBα) were determined by Western Blot. Results No mice died in the NS control group at 12 hours. In the treatment group, the mortality at 12 hours was significantly lower than that in the model group (10% vs. 80%, P < 0.01). The levels of tryptase and IgE in the model group were significantly higher than those in the NS control group [tryptase (μg/L): 1.53±0.28 vs. 0.91±0.23, IgE (μg/L): 33.3±3.1 vs. 21.3±1.9, both P < 0.01], both levels in the treatment group were significantly lower than those in the model group [tryptase (μg/L): 1.31±0.26 vs. 1.53±0.28, IgE (μg/L): 25.6±2.2 vs. 33.3±3.1, both P < 0.05]. The levels of TNF-α, IL-1 and IL-6 in the model group were significantly higher than those in the NS control group [TNF-α (ng/L): 35.3±4.7 vs. 16.4±3.5, IL-1 (ng/L): 13.8±3.3 vs. 4.2±1.8, IL-6 (ng/L): 15.3±4.8 vs. 5.5±2.1, all P < 0.01]. The serum inflammatory factors of the treatment group were significantly higher than those of the model group [TNF-α (ng/L): 26.1±4.3 vs. 35.3±4.7, IL-1 (ng/L): 7.2±2.7 vs. 13.8±3.3, IL-6 (ng/L): 8.8±3.8 vs. 15.3±4.8, all P < 0.05]. It was shown by Western Blot results that there was no significant difference in p65 protein expression of the lung tissue among the three groups. In the NS control group, the expression of p-p65 protein in the nucleus of the lung tissue was extremely low but was significantly increased in the model group, and p-p65 protein in the treatment group was significantly decreased as compared with that in the model group. The expression tendency of p-IκBα protein was consistent with that of p-p65. Conclusion Epinephrine combined with acupuncture plays a therapeutic role in mice with anaphylactic shock by inhibiting the activation of NF-κB signaling pathway.
ABSTRACT
AIM:To investigate the regulatory effect of nuclear factor-κB (NF-κB) inhibitor, pyrrolidine di-thiocarbamate (PDTC), on nerve function and neural cell apoptosis in rats after intracerebral hemorrhage (ICH).METH-ODS:SPF Sprague-Dawley rats were randomly divided into 4 groups with 6 rats in each group: sham group, ICH group, PDTC at low concentration (Plow) group and PDTC at high concentration (Phigh) group.Autologous blood injection was used to establish ICH model.After 2 h of surgery, the rats in Plow group and Phigh group were intraperitoneal injected with PDTC at 100 mg/kg and 200 mg/kg, respectively , while rats in sham group and ICH group were injected with the same volume of saline .The neurological function score was classified with modified Longa grading method .TUNEL assay was used to detected the neural cell apoptosis , and the content of malondialdehyde ( MDA) and the activity of superoxide dis-mutase (SOD) were measured.Furthermore, the protein levels of p-P65 and cleaved caspase-3 in brain tissues were deter-mined by Western blot .RESULTS: Compared with sham group , the rats in ICH group had higher neurological function score (P<0.05).After treatment with PDTC, the neurological function score was decreased (P<0.05), but no signifi-cant difference between P low group and P high group was observed .Compared with sham group , the number of apoptotic cells in ICH group was increased ( P<0.05 ) .After treatment with PDTC , the neural cell apoptosis was restrained , and the number of apoptotic cells in Phigh group was lower than that in Plow group (P<0.05).Compared with sham group, the con-tent of MDA was increased and the activity of SOD was decreased in ICH group (P<0.05).After treatment with PDTC, the content of MDA was decreased while the activity of SOD was increased , and the variation trend was more obvious in Phigh group (P<0.05).Compared with sham group, the protein levels of p-P65 and cleaved caspase-3 in ICH group were increased (P<0.05).After treatment with PDTC, the protein levels of p-P65 and cleaved caspase-3 were decreased, and those in Phigh group were lower than those in P low group.CONCLUSION: NF-κB inhibitor PDTC plays a role in the se-condary brain injury after ICH , and the protective effect increases at the higher dose .The mechanism may be related to re-ducing MDA content and increasing SOD activity , and further inhibiting neural cell apoptosis .
ABSTRACT
Objective To explore the effects of Buyang Huanwu decoction(BYHWD)on expressions of nuclear factor-κB p65(NF-κBp65)and its inhibitor( I-κB)in signal transduction of NF-κB in brain tissue of rats with focal cerebral ischemia injury. Methods 180 Sprague-Dawley(SD)rats were randomly divided into normal group,sham-operated group,model group,pynolidine dithiocarbamate(PDTC)group,minocycline(MC)group and BYHWD treatment group,each group 30 rats. The rats of PDTC group were given PDTC 100 mg?kg-1?d-1 by intra-peritoneal injection. In MC group,MC was given by filling the stomach,the dose was 2.35 g?kg-1?d-1,the drug solution was prepared by adding the distilled water,and the total volume of drug solution to fill the stomach was kept at the same volume in various groups,thus the concentration of the drug was different. In BYHWD group,BYHWD was given,the dose was reduced to 5 g?kg-1?d-1 according to the body surface area dose conversion formula about people and animals. In sham-operated group and model group,the distilled water was given in the same volume as other drug solution. The protein expression levels of NF-κBp65 and I-κB in ischemic tissues were examined by using immunohistochemical method on the time points 7,14 and 21 days after treatment in each group. Results Compared with model group, the cell numbers with expression of NF-κBp65 in PDTC group,MC group and BYHWD group were significantly decreased along with the prolongation of therapy time,the decrease in number was more and more,until 21 days,it reached the valley level(cell/400 times HP:44.00±6.91,45.33±6.55,18.67±2.14 vs. 126.00±5.78,all P0.05). After treatment for 7 days,the number of cells with positive expression of I-κB protein in BYHWD group was less than that in MC group(cell/400 times HP:55.00±3.40 vs. 72.50±4.29,P0.05),and after treatment for 21 days,the number in BYHWD group was significantly higher than that in MC group(88.83±4.95 vs. 71.17±7.16, P<0.05). Conclusion BYHWD can regulate the expressions of inflammatory cytokine I-κB and NF-κB in signal transduction of NF-κB in ischemic brain tissue to inhibit the inflammatory reaction,thus it has the protective effect on cerebral ischemia.