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Objective:To investigate the effect of Xiao Xumingtang combined with super-acupuncture along governor meridian on autophagy-related protein nuclear factor-kappa B p65 (NF-κB p65) in cerebral ischemia-reperfusion model rats, so as to study the relationship between autophagy-related protein NF-κB p65 and brain protection mechanism, and look for the best intervention time point of acupuncture. Method:A total of 152 adult SD male rats were randomly divided into sham operation group, model group, high-dose Xiao Xumingtang group (high-dose drug group), low-dose Xiao Xumingtang group (low-dose drug group) and acupuncture group. There were seven groups including high-dose Xiao Xumingtang + acupuncture group (high acupuncture group) and low-dose Xiao Xumingtang + acupuncture (low acupuncture group). Model group, high-dose drug group, low-dose drug group, and acupuncture group were divided into 4 subgroups according to 30 minutes, 2, 4， 6 h of ischemia-reperfusion, with 6 animals in each group. After successful modeling, according to Zea Longa's neural function score, eligible rats were included into the corresponding groups. The sham operation group only received carotid artery dissection, the model group was only modeled without any treatment, high and low-dose Xiao Xumingtang groups were calculated based on the body surface area of the animal and given 60 g·kg-1·d-1 and 15 g·kg-1·d-1 drug by gavage for treatment, acupuncture was performed to smooth governor meridian and regulate the mind. After 14 days of consecutive treatment, neurological function was scored. Western blot was used to detect the expression of autophagy-related protein NF-κB p65 in rat brain tissue. Result:Compared with the sham operation group, the neurological impairment scores of the model group, the high-dose drug group, the low-dose drug group, the acupuncture group, the high acupuncture group, and the low acupuncture group were significantly increased at each time point (P<0.01). The neurological impairment scores were significantly lower at each time point than those of the high-dose drug group, low-dose drug group, acupuncture group, high acupuncture group, and low acupuncture group (P<0.01), compared with the sham operation group, NF-κB p65 in model group, high-dose drug group, low-dose group, acupuncture group, high acupuncture group and low acupuncture group was significantly increased in the brain tissue at each time point (P<0.01), compared with the model group, the expression of NF-κB p65 protein in the brain tissue of model group, high-dose drug group, low-dose group, acupuncture group, high acupuncture group and low acupuncture group was decreased at each time point (P<0.05), particular in the high acupuncture group (P<0.01). Conclusion:Xiao Xumingtang combined with ultra-early acupuncture along governor meridian can significantly alleviate neurological impairment in rats with cerebral ischemia-reperfusion model. Xiao Xumingtang combined with ultra-early acupuncture along governor meridian can inhibit cerebral ischemia-reperfusion model rats. The activity of autophagy-related protein NF-κB P65 protects the brain function. There is no significant difference in the brain protective effect of Xiao Xumingtang combined with ultra-early acupuncture along governor meridian within 6 hours.
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Objective: To study the effect of modified Erchentang on expressions of Toll-like receptor 4 (TLR4), myeloid differentiation factor (MyD88) and nuclear factor-κB (NF-κB) genes in the lung tissue homogenate of rats with chronic obstructive pulmonary disease (COPD). Method: Forty SD rats were randomly divided into normal group, model group, modified Erchentang group and EVP4593 (NF-κB inhibitor) group. Rat COPD models were prepared through cigarette smoke and tracheal dripping with lipopolysaccharide (LPS). After the modeling, normal and model groups were intragastrically given normal saline solution, EVP4593 group was given EVP4593(1 mg · kg-1) through subcutaneous injection, and modified Erchentang group was given corresponding herbal drugs intragastrically (10 g · kg-1) for 14 days. The levels of high mobility group box 1(HMGB1), chemokines CXCL-2, CXCL-3 and monocyte chemoattractant protein-1 (MCP-1) in rats serum were detected by enzyme-linked immunosorbent assay in rats serum. The expressions of Toll-like receptors 4(TLR4), myeloid differentiation factor (MyD88) and nuclear factor-κB p65 (NF-κB p65) mRNA were detected by Real-time fluorescence quantitative PCR (Real-time PCR) method. Western blot were used to detect the levels of TLR4, MyD88, NF-κB p65 and p-NF-κB p65 protein. Immunohistochemistry (IHC) method was used to detect the localization and expressions of TLR4, MyD88 and p-NF-κB p65 protein in the lung tissue. Result: The mRNA and protein expressions of TLR4, MyD88 and NF-κB p65 were increased significantly (PPκB p65 mRNA and protein were decreased significantly (PConclusion: Modified Erchentang may inhibit the inflammatory response of COPD effectively. The mechanism may be related to the inhibition of the expressions of the signal molecule genes involved in the TLR4/MyD88/NF-κB pathway and the reduction of the release of HMGB1, CXCL-2, CXCL-3 and MCP-1.
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Objective To observe the effect of electroacupuncture(EA)at Baihui(GV20)and Siguan(Hegu/LI4 and Taichong/LR3)of affected side on expression of Tax1-binding protein 1(TAX1BP1)in cerebral cortex in focal cerebral ischemia-reperfusion rats,so as to in-vestigate its protective mechanism in inhibiting nuclear factor kappa B(NF-κB)signaling pathway and promoting neurobehavioral recovery. Methods A total of 105 Sprague-Dawley rats were randomly assigned into sham group,model group and EA group.Each group was ran-domly assigned into reperfusion six hours,twelve hours,24 hours,48 hours,72 hours groups after two hours of ischemia.The model was es-tablished by right middle cerebral artery occlusion and reperfusion.EA group received electroacupuncture at Baihui and left Siguan(Hegu and Taichong)acupoints.Neurobehavioral evaluation,TAX1BP1 protein expression,TAX1BP1 positive cell count,zink finger protein A20 expression, and nuclear NF-κB p65 protein expression were tested in each group. Results There was no neurological deficit in the sham group. Compared with the model group, the neurological scores at 48 hours, 72 hours after reperfusion decreased in EA group (P<0.05). Compared with the control group,the TAX1BP1 expression at twelve hours,24 hours and 48 hours after reperfusion increased in the model group(P<0.05),and further increased at twelve hours,24 hours,48 hours and 72 hours after reperfusion in EA group(P<0.05),and peaked at 24 hours after reperfusion.Compared with the sham group,the expression of A20 and NF-κB p65,and the number of TAX1BP1 positive cells increased in the model group(P<0.05),and the expression of A20 and the number of TAX1BP1 positive cells further increased,(P<0.05)and the expression of NF-κB p65 decreased in EA group(P<0.05)at 24 hours after reperfusion.Immunofluorescence labeling indicat-ed that TAX1BP1 protein primarily expressed in the cytoplasm,TAX1BP1 protein and A20 protein co-expressed in the cytoplasm.Immuno-histochemistry showed indicated that NF-κB p65 mainly expressed in the nucleus in the model groupr,and mainly expressed in the cyto-plasm in the EA group.Conclusion Electroacupuncture could significantly inhibit neuronal NF-κB signaling pathway and promote neurobe-havioral recovery in focal cerebral ischemia-reperfusion,which may related with up-regulating TAX1BP1 protein expression.
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Objective To investigate the effect of different doses of dexmedetomidine (DEX) on myocardial ischemia-reperfusion injury (MIRI) in rats.Methods Seventy-five SD male rats weighing between 250 and 300g were divided into sham group,MIRI group,small-dose DEX group,medium-dose DEX group and high-dose group according to the random number table,with 15 rats per group.Threading the left anterior descending coronary artery was done only in sham group,but the MIRI model was produced in the rest groups by ligation of the artery for 30 minutes followed by 120 minutes of reperfusion.Fifteen minutes before the ligation,small-,medium-and high-dose DEX groups were injected 2.5,5 and 10 μg · kg-1 · h-1 of DEX respectively until the end of reperfusion.Instead,an equal volume of normal saline was given in sham and MIRI groups.At the end of reperfusion,five rats in each group were used to determine the myocardial infarct size,and arterial blood samples and myocardial tissues from ten rats in each group were used to measure serum levels of interleukin-1β (IL-β) and serum tumor necrosis factor-α (TNF-α),expression of myocardial nuclear factor kappa B p65 (NF-κB p65) and change of myocardial pathomorphology.Results Myocardial infract size,degree of myocardial pathomorphology structure damage,serum levels of IL-1 β and TNF-αt and expression of myocardial NF-κB p65 in sham group were significantly lower in sham group than other groups (P ＜ 0.05).Above mentioned parameters in small-,medium-and high-dose DEX groups were all significantly decreased compared to MIRI group (P ＜ 0.05),and the decrease was most significant in medium-dose DEX group (P ＜ 0.05).Conclusions DEX can attenuate the MIRI in rats and the possible mechanism is suppressing the release of NF-κB p65,which can reduce serum pro-inflammatory cytokines like TNF-α and IL-1β.And mediumdose DEX exhibits better protective effect.
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Objective To investigate the effect of recombinant human growth hormone(rhGH)on the protein expression of NF-?B P65 in ischemic reperfusion injury of rat liver.Methods One hundred male rats models were constructed by Pringle's method and were randomly divided into two groups: the rhGH group and the control group.In rhGH group, the rats were injected(0.2IU/100g weight)seven days before the ischemic reperfusion injury,while in control group,rats was replaced by normal sodium.The levels of protein expression of NF-?B P65 were detected by Western blotting and immunohistochemistry.Results Compared with control group,the expression of NF-?B P65 and NF-?B activity(P