[Effect of moxibustion on Nogo/neurotrophin receptor signaling pathway in rats with cerebral ischemia/reperfusion injury].

OBJECTIVE: To observe the effect of moxibustion treatment on the expression of Nogo-A, Nogo receptor (NgR), neurotrophin receptor p75 (p75NTR) and leucine rich repeat and Ig domain containing 1 (Lingo-1) in brain tissue of rats with cerebral ischemia/reperfusion injury (CI/RI), so as to analyze its mechanism underlying improvement of CI/RI. METHODS: Male SD rats were randomly divided into sham operation group (16 rats), model group (17 rats), NEP1-40 (extracellular peptide residues 1-40, a blocker targeting NgR) group (model+blocker, 17 rats) and moxibustion group (model+moxibustion, 17 rats). The CI/RI model was established by occlusion of the left middle cerebral artery (MCAO). Moxibustion was applied to "Baihui"(GV20), right "Quchi"(LI11) and "Zusanli"(ST36) for 20 min, once a day for 14 days, with 2 days' rest after the top 7 days' intervention. For rats of the NEP1-40 group, 30 µL PBS containing 18 µg NEP 1-40 was injected into the epidural inferior vena (L5-S1) via a polyvinyl chloride conduit. The neurological deficit state in each group was evaluated by Longa's 5-point scale and Feeney's 7-point scale of beam walking test (BWT). The cerebral infarct volume was assessed by 2,3,5-triphenyltetrazole chloride staining. The brain tissue between the central anterior and posterior sulcus was taken for observing the expression of NgR and Lingo-1 by fluorescence double-label method, and for determining the expression levels of Nogo-A, NgR, p75NTR and Lingo-1 mRNAs and proteins by real-time quantitative PCR and Western blot, respectively. RESULTS: After modeling, the Longa's score, infarct volu-me percent, expression levels of Nogo-A, NgR, Lingo-1 and p75NTR mRNAs and proteins were significantly increased (P<0.01) and BWT score was obviously decreased (P<0.01) in the model group relevant to the sham operation group. In comparison with the model group, the increase of Longa's score, infarct volume percentage, expression levels of Nogo-A, NgR, Lingo-1 and p75NTR mRNAs and proteins and decrease of BWT score in NEP1-40 and moxibustion groups were reversed (P<0.01) except Nogo-A protein in the NEP1-40 group. The effect of moxibustion was significantly superior to that of blocker NEP1-40 in redu-cing the infarct volume percentage, and down-regulating the expression of Nogo-A mRNA and protein, p75NTR mRNA and protein, NgR and Lingo-1 proteins (P<0.01, P<0.05). CONCLUSION: Moxibustion, similar to blocker NEP1-40 of NgR, can improve neurological dysfunction in CI/RI rats, which may be related to its functions in reducing cerebral infarction and down-regulating the activity of Nogo/neurotrophin receptor signaling pathway.

[Effect of scalp acupuncture stimulation on cerebral infarct volume and expression of IL-10, IL-6 and IL-1ß in para-hippocampal gyrus in acute ischemic cerebrovascular disease rats].

OBJECTIVE: To observe the influence of scalp acupuncture on cerebral infarct size and expression of IL-10, IL-6, and IL-1ß in the para-hippocampal gyrus in acute ischemic cerebrovascular disease(AICD) rats, so as to investigate its mechanisms underlying improvement of AICD. METHODS: Forty-eight male SD rats were randomly allocated to normal control (control), AICD model, medication, and scalp acupuncture groups (n＝12 per group). The AICD model was established by occlusion of the middle cerebral artery (MCAO). Rats of the medication group received intraperitoneal injection of Ammonium 1-Pyrrolidinedithiocarbamate (APDC, 100 mg•kg－1•d－1), once daily for 7 days. Scalp acupuncture stimulation was applied to bilateral "Dingnieqianxiexian" (MS6) once daily for 7 days. Before and after intervention, the neurologic deficit score (NDS) and the neurological score (NS) were evaluated according to Longa's and Schäbitz's methods, respectively. At the end of the intervention, the para-hippocampal gyrus and whole brain were collected respectively. The expression levels of IL-10, IL-6 and IL-1ß in the para-hippocampal gyrus tissue were detected by immunohistochemistry, and the cerebral infarct volume of the brain was detected by triphenyltetrazollium chloride (TTC) staining after sectioning. RESULTS: Following modeling, the NDS, NS and the expression of IL-10, IL-6 and IL-1ß in para-hippocampal gyrus were significantly increased in the model group compared with the control group (P<0.01). After the intervention, the NDS, NS and infarct volume, and the expression levels of IL-6 and IL-1ß in the para-hippocampal gyrus were significantly decreased in both medication and scalp acupuncture groups compared with the model group (P<0.05), and the expression of IL-10 was further obviously up-regulated in the scalp acupuncture group (P<0.05) rather than in the medication group (P>0.05). The effect of scalp acupuncture was obviously superior to that of medication in up-regulating IL-10 expression level (P<0.05). No significant differences were found between the medication and scalp acupuncture groups in the levels of NDS, NS, infarct volume, IL-6 and IL-1ß proteins (P>0.05). CONCLUSION: Scalp acupuncture can improve neurological function and reduce infarct volume in AICD rats, which may be associated with its function in up-regulating the expression of IL-10 and in inhibiting the expression of IL-6 and IL-1ß to reduce inflammation reaction.

Effect of scalp acupuncture stimulation on cerebral infarct volume and expression of IL-10, IL-6 and IL-1β in para-hippocampal gyrus in acute ischemic cerebrovascular disease rats / 针刺研究

OBJECTIVE: To observe the influence of scalp acupuncture on cerebral infarct size and expression of IL-10, IL-6, and IL-1β in the para-hippocampal gyrus in acute ischemic cerebrovascular disease(AICD) rats, so as to investigate its mechanisms underlying improvement of AICD. METHODS: Forty-eight male SD rats were randomly allocated to normal control (control), AICD model, medication, and scalp acupuncture groups (n＝12 per group). The AICD model was established by occlusion of the middle cerebral artery (MCAO). Rats of the medication group received intraperitoneal injection of Ammonium 1-Pyrrolidinedithiocarbamate (APDC, 100 mg•kg－1•d－1), once daily for 7 days. Scalp acupuncture stimulation was applied to bilateral "Dingnieqianxiexian" (MS6) once daily for 7 days. Before and after intervention, the neurologic deficit score (NDS) and the neurological score (NS) were evaluated according to Longa's and Schäbitz's methods, respectively. At the end of the intervention, the para-hippocampal gyrus and whole brain were collected respectively. The expression levels of IL-10, IL-6 and IL-1β in the para-hippocampal gyrus tissue were detected by immunohistochemistry, and the cerebral infarct volume of the brain was detected by triphenyltetrazollium chloride (TTC) staining after sectioning. RESULTS: Following modeling, the NDS, NS and the expression of IL-10, IL-6 and IL-1β in para-hippocampal gyrus were significantly increased in the model group compared with the control group (P0.05). The effect of scalp acupuncture was obviously superior to that of medication in up-regulating IL-10 expression level (P0.05). CONCLUSION: Scalp acupuncture can improve neurological function and reduce infarct volume in AICD rats, which may be associated with its function in up-regulating the expression of IL-10 and in inhibiting the expression of IL-6 and IL-1β to reduce inflammation reaction.

Neuroprotective effect of baicalin on focal cerebral ischemia in rats.

Baicalin, a flavonoid compound from the root of the herb Scutellaria baicalensis Georgi, has been widely used to treat patients with inflammatory disease. The aim of this study was to assess the efficacy of baicalin in a rat model of focal cerebral ischemia. Adult male Sprague-Dawley rat models of cerebral artery occlusion were established and then randomly and equally divided into three groups: ischemia (cerebral ischemia and reperfusion), valproic acid (cerebral ischemia and reperfusion + three intraperitoneal injections of valproic acid; positive control), and baicalin (cerebral ischemia and reperfusion + intraperitoneal injection of baicalin for 21 days). Neurological deficits were assessed using the postural reflex test and forelimb placing test at 3, 7, 14, and 21 days after ischemia. Rat cerebral infarct volume was measured using 2,3,5-triphenyltetrazolium chloride (TTC) staining method. Pathological change of ischemic brain tissue was assessed using hematoxylin-eosin staining. In the baicalin group, rat neurological function was obviously improved, cerebral infarct volume was obviously reduced, and the pathological impairment of ischemic brain tissue was obviously alleviated compared to the ischemia group. Cerebral infarct volume was similar in the valproic acid and baicalin groups. These findings suggest that baicalin has a neuroprotective effect on cerebral ischemia.

[Acupuncture Reduces Acute Cerebral Ischemia by Suppressing Local and Peripheral Inflammation and Regulating Expression of Cerebral Protein Silent Information Regulator Factor 2-related Enzyme 1 and Nuclear Factor-κB in Rats].

OBJECTIVE: To observe the effect of acupuncture on the expression of silent information regulator factor 2-related enzyme 1 (SIRT 1) and nuclear factor-κB (NF-κB) in rats with acute cerebral ischemia (ACI), so as to explore its mechanisms underlying improvement of ACI. METHODS: One hundred female SD rats were randomly divided into 5 groups: normal control (normal), sham-operation (sham), model, non-acupoint and acupoint, with 20 rats in each. The ACI model was established by occlusion (electric coagulation) of the middle cerebral artery after craniotomy. "Baihui" (GV 20) and "Shuigou" (GV 26) or non-acupoints were punctured with filiform needles which were retained for 30 min after rotating for 1 min. The treatment was conducted once after modeling and 24 h thereafter. The cerebral infarct volume was measured after 2,3,5-triphenyltetrazolium chloride (TTC) staining. The contents of interleukin-1 ß (IL-1 ß), IL-6 and IL-8 in the serum and ischemic brain tissue were detected by radioimmunoassay, and the expression of protein SIRT 1 and NF-κB p 65 in the ischemic brain tissue was detected by immunoblotting. RESULTS: The brain infarction volume was obvious in the model group in comparison with the normal group (P<0.01), but was markedly reduced in the acupoint group relevant to the model group (P<0.05). The contents of IL-1 ß, IL-6 and IL-8 in the serum and ischemic brain tissues, and the expression level of cerebral NF-κB p 65 protein were significantly increased (P<0.01), and the expression of cerebral SIRT 1 was obviously decreased in the model group compared with the normal group (P<0.01). Following acupuncture intervention, the increased IL-1 ß, IL-6 and IL-8 contents and NF-κB p 65 expression were considerably down-regulated, and the decreased SIRT 1 expression was markedly up-regulated in the acupuncture acupoint group (P<0.05). No significant changes were found in the levels of all the indexes mentioned above between the normal and sham groups (P>0.05). CONCLUSION: Acupuncture intervention at acupoints can reduce the ischemic infarction volume in ACI rats, which may be associated with its effects in down-regulating the levels of IL-1 ß, IL-6 and IL-8 in the serum and ischemic brain tissue, and in regulating cerebral SIRT 1/NF-κB signaling.

Acupuncture Reduces Acute Cerebral Ischemia by Suppressing Local and Peripheral Inflammation and Regulating Expression of Cerebral Protein Silent Information Regulator Factor 2-related Enzyme 1 and Nuclear Factor-κB in Rats / 针刺研究

OBJECTIVE: To observe the effect of acupuncture on the expression of silent information regulator factor 2-related enzyme 1 (SIRT 1) and nuclear factor-κB (NF-κB) in rats with acute cerebral ischemia (ACI), so as to explore its mechanisms underlying improvement of ACI. METHODS: One hundred female SD rats were randomly divided into 5 groups: normal control (normal), sham-operation (sham), model, non-acupoint and acupoint, with 20 rats in each. The ACI model was established by occlusion (electric coagulation) of the middle cerebral artery after craniotomy. "Baihui" (GV 20) and "Shuigou" (GV 26) or non-acupoints were punctured with filiform needles which were retained for 30 min after rotating for 1 min. The treatment was conducted once after modeling and 24 h thereafter. The cerebral infarct volume was measured after 2,3,5-triphenyltetrazolium chloride (TTC) staining. The contents of interleukin-1 β (IL-1 β), IL-6 and IL-8 in the serum and ischemic brain tissue were detected by radioimmunoassay, and the expression of protein SIRT 1 and NF-κB p 65 in the ischemic brain tissue was detected by immunoblotting. RESULTS: The brain infarction volume was obvious in the model group in comparison with the normal group (P0.05). CONCLUSION: Acupuncture intervention at acupoints can reduce the ischemic infarction volume in ACI rats, which may be associated with its effects in down-regulating the levels of IL-1 β, IL-6 and IL-8 in the serum and ischemic brain tissue, and in regulating cerebral SIRT 1/NF-κB signaling.

[Effects of electroacupuncture preconditioning on cerebral infarct volume and contents of TNF-α, IL-10 in serum of rats after cerebral ischemia-reperfusion injury].

OBJECTIVE: To explore the effect of electroacupuncture(EA) preconditioning on cerebral infarct volume and the contents of TNF-α, IL-10 in serum of rats with cerebral ischemia-reperfusion injury. METHODS: Thirty-six rats were randomly divided into a sham operation group, a model group and an EA preconditioning group, 12 rats in each group, which were further divided into 12 h and 24 h after reperfusion subgroups, 6 rats in each one. EA was used before model establishment for 2 weeks in the EA preconditioning group. The model of cerebral ischemia-reperfusion injury in rats was established with modified Longa suture method. 12 h and 24 h after reperfusion, the degree of neurological deficit was assessed by the modified behavioral scoring scale; the cerebral infarct volume was measured by TTC method and the contents of TNF-α, IL-10 in serum were detected by ELISA method. RESULTS: Compared with the model group, the neurological severity scores in the EA preconditioning group significantly reduced 12 h and 24 h after reperfusion (both P<0.05), the cerebral infarct volume in the EA preconditioning group significantly reduced 12 h and 24 h after reperfusion (both P<0.05). Compared with the sham operation group, the serum TNF-α, IL-10 contents in the model group increased 12 h and 24 h after reperfusion (both P<0.05). Compared with the model group, the serum TNF-α content reduced, while the serum IL-10 content increased in the EA preconditioning group 12 h after reperfusion (both P<0.05). Compared with the model group, the serum TNF-α, IL-10 contents reduced in the EA preconditioning group 24 h after reperfusion (both P<0.05). CONCLUSION: EA preconditioning can improve neurological deficit, reduce cerebral infarct volume after cerebral ischemia-reperfusion injury in rats. The mechanism may be related to the regulation of EA on the dynamic balance between pro-inflammatory cytokine TNF-α and anti-inflammatory cytokine IL-10 in peripheral blood of cerebral ischemia-reperfusion injury in acute phase, thus alleviate acute cerebral ischemia-reperfusion inflammatory response.

Effects of electroacupuncture preconditioning on cerebral infarct volume and contents of TNF-α, IL-10 in serum of rats after cerebral ischemia-reperfusion injury / 中国针灸

<p><b>OBJECTIVE</b>To explore the effect of electroacupuncture(EA) preconditioning on cerebral infarct volume and the contents of TNF-α, IL-10 in serum of rats with cerebral ischemia-reperfusion injury.</p><p><b>METHODS</b>Thirty-six rats were randomly divided into a sham operation group, a model group and an EA preconditioning group, 12 rats in each group, which were further divided into 12 h and 24 h after reperfusion subgroups, 6 rats in each one. EA was used before model establishment for 2 weeks in the EA preconditioning group. The model of cerebral ischemia-reperfusion injury in rats was established with modified Longa suture method. 12 h and 24 h after reperfusion, the degree of neurological deficit was assessed by the modified behavioral scoring scale; the cerebral infarct volume was measured by TTC method and the contents of TNF-α, IL-10 in serum were detected by ELISA method.</p><p><b>RESULTS</b>Compared with the model group, the neurological severity scores in the EA preconditioning group significantly reduced 12 h and 24 h after reperfusion (both<0.05), the cerebral infarct volume in the EA preconditioning group significantly reduced 12 h and 24 h after reperfusion (both<0.05). Compared with the sham operation group, the serum TNF-α, IL-10 contents in the model group increased 12 h and 24 h after reperfusion (both<0.05). Compared with the model group, the serum TNF-α content reduced, while the serum IL-10 content increased in the EA preconditioning group 12 h after reperfusion (both<0.05). Compared with the model group, the serum TNF-α, IL-10 contents reduced in the EA preconditioning group 24 h after reperfusion (both<0.05).</p><p><b>CONCLUSION</b>EA preconditioning can improve neurological deficit, reduce cerebral infarct volume after cerebral ischemia-reperfusion injury in rats. The mechanism may be related to the regulation of EA on the dynamic balance between pro-inflammatory cytokine TNF-α and anti-inflammatory cytokine IL-10 in peripheral blood of cerebral ischemia-reperfusion injury in acute phase, thus alleviate acute cerebral ischemia-reperfusion inflammatory response.</p>

Influence of Buyanghuanwu Decoction on Cerebral Infarct Volume and Pathological Changes after Cerebral Ischemia and Reperfusion / 中国中医药信息杂志

Objective To investigate the influence of Buyanghuanwu (BYHW) Decoction on cerebral infarct volume and pathological changes after cerebral ischemia and reperfusion. Methods The model of focal cerebral ischemia and reperfusion was made by thread embolism method. Healthy male SD rats were divided randomly into sham-operated group, model group and BYHW Decoction group. Infarct volume was detected by triphenyltetrazolium chloride (TTC) staining and pathological changes were observed microscopically in HE stained sections. Results In BYHW Decoction group, infarct volume was significantly smaller than that in model group (P