[Electroacupuncture improves intestinal function by down-regulating expression of 5-HT1AR and c-fos proteins in the hypothalamus and colon tissues via brain-gut axis in rats with functional diarrhea].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on expression of 5-HT1A receptor (5-HT1AR) and c-fos proteins in the hypothalamus and colon tissues in functional diarrhea (FD) rats so as to explore its underlying mechanisms in improving intestinal function via brain-gut axis. METHODS: A total of 40 SD rats were randomized into blank control, model, EA Tianshu (ST25)-Dachangshu (BL25, ST25-BL25) and EA Quchi (LI11)- Shangjuxu (ST37, LI11-ST37) groups. The FD model was established by gavage of Folium Sennae (10 mg/kg) and constraint immobilization once daily for 29 days. EA (10 Hz/50 Hz, 1.5 mA) was applied to bilateral ST25 and BL25 in EA ST25-BL25 group, and bilateral LI11 and ST37 in EA LI11-ST37 group for 20 min, once daily for successive 10 days. The expression of 5-HT1AR and c-fos protein in the hypothalamus and colon tissues was determined by using Western blot. The state of stool was recorded for calculating the loose stool rate and diarrhea index. RESULTS: After modeling, the loose stool rate, diarrhea index and expression levels of 5-HT1AR and c-fos proteins in the colon and hypothalamus tissues were obviously increased in the model group in contrast with the blank control group(P<0.01，P<0.05). Following EA intervention, the loose stool rate and diarrhea index, the expression levels of 5-HT1AR and c-fos proteins in the hypothalamus and colon in the EA ST25-BL25 group, and the expression of 5-HT1AR in the colon in the EA LI11-ST37 group were significantly down-regulated relevant to the model group (P<0.01，P<0.05). No significant changes were found in loose stool rate and diarrhea index，and the expression levels of hypothalamic 5-HT1AR and hypothalamic and colonic c-fos proteins in the EA LI11-ST37 group (P>0.05). The expression levels of 5-HT1AR protein in the hypothalamus and c-fos protein in the hypothalamus and colon in the EA LI11-ST37 group were significantly up-regulated relevant to the EA ST25-BL25 group (P<0.05). CONCLUSION: EA of ST25-BL25 can down-regulate expression of 5-HT1AR and c-fos proteins in the hypothalamus and colon tissue in FD rats, which may contribute to its function in improving symptoms of FD possibly via brain-gut axis.

Silibinin exerts antidepressant effects by improving neurogenesis through BDNF/TrkB pathway.

Classic antidepressants benefit depression patients partially by improving neurogenesis and/or brain-derived neurotrophic factor (BDNF)/TrkB pathway which were impaired in depression. In this study, we demonstrated that Silibinin (SLB), a polyphenolic flavanoid from Silybum marianum, ameliorated reserpinized mouse depressant-like behaviors. The antidepressants of SLB administration was associated with increased neural stem cells (NSCs) proliferation and further confirmed in BDNF/TrkB signaling transduction. SLB treatment reversed the decreased expression levels of BDNF and its receptor TrkB, and the reduced activation of downstream target proteins including phosphorylated extracellular-regulated protein kinase (p-ERK) and phosphorylated cAMP-response element binding protein (p-CREB) in depressived hippocampus. Furthermore, intracerebroventricular injection of GNF5837, a TrkB antagonist, abrogated antidepressant-like effects of SLB in mice along with the improved NSC proliferation, as well as enhanced levels of p-ERK and p-CREB in mice hippocampus. Taken together, these results suggest that SLB may exert antidepressant effects through BDNF/TrkB signaling pathway to improve NSC proliferation in acute depression.

[Effect of "Xiusanzhen" on hippocampal muscarinic cholinergic receptor activity in Alzheimer disease rats].

OBJECTIVE: To explore the effect of "Xiusanzhen" [electroacupuncture (EA) of bilateral "Yingxiang" (LI 20) and "Yintang" (EX-HN 3)] on activities of hippocampal muscarinic cholinergic receptor (mAChR) and the involved neural path in Alzheimer Disease(AD)rats. METHODS: Forty Sprague Dawley rats were randomly divided into normal control, model, olfactory nerve severing (ONS)-EA of "Xiusanzhen" (ONS-EA) and EA of "Xiusanzhen" (EA) groups. AD model was established by intra-hippocampal injection (AP: 3.5 mm, L:2 mm, H: -2.8 mm) of Abeta(1-40) starch-like peptide (10 microg/2 microL) under the aid of a microsyringe installed in a brain stereotaxis instrument. For rats of the ONS-EA group, the olfactory nerve was severed by using a surgical knife after drilling a hole on the skull (5 mm anterior to the bregma, L, R: 2 mm). The mAChR density, and its maximum binding capacity (Bmax) and dissociation constant (Kd) of the hippocampus tissue were measured by using radio-ligand binding analysis and Lowry's microamount protein assay. RESULTS: In comparison with the normal control group, the hippocampal mAChR density and its Bmax in the model group were decreased remarkably (P < 0.05), while the Kd of M-receptor in the model group was increased remarkably (P < 0.05). In comparison with the model group, hippocampal mAChR density and its Bmax in the EA group were up-regulated obviously (P < 0.05), while the Kd of mAChR in the EA group was down-regulated significantly (P < 0.05). No significant differences were found between the model and ONS-EA groups in mAChR density and its Bmax and Kd (P > 0.05). CONCLUSION: "Xiusanzhen"-EA can effectively up-regulate hippocampal mAChR density and Bmax and down-regulate Kd of M-receptor of hippocampus in AD rats, which is dependent on the intact olfactory nerve pathway.

[Effect of "Xiusanzhen"on expression of hippocampal Bcl-2 and Bax proteins in Alzheimer disease rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) of "Xiusanzhen" [3 acupoints, i.e., bilateral "Yingxiang" (LI 20) and "Yintang" (EX-HN 3)] on expression of B-cell lymphoma/leukemia-2 (Bcl-2, an antiapoptosis protein) and Bax (a protein for promoting apoptosis) in the hippocampus in Alzheimer disease (AD) rats, so as to explore its clinical mechanisms underlying relieving AD. METHODS: A total of 40 Sprague Dawley rats were randomly divided into normal control, AD model, EA and olfactory nerve transaction(ONT) + EA groups, with 10 rats in each. AD model was established by injection of beta-amyloid (Abeta)(1-40) (2 microL containing 10 microg) into the hippocampus (Bregma: 3.5 mm, L: 2.0 mm, H: -2.8 mm below dura), and the ON was sectioned by using a fine needle. EA (80-100 Hz, 1-3 mA) was applied to bilateral LI 20 and EX-HN 3 for 10 min, once daily (except Saturdays and Sundays) for 6 weeks. The expression of Bcl-2 and Bax of hippocampus was measured by immunohisto-chemistry. RESULTS: Compared with the normal control group, the expression of Bcl-2 of hippocampus was significantly lower in the AD model group (P<0.05), and that of Bax in AD model group was much higher (P<0.01). In comparison with the model group, the expression of hippocampal Bcl-2 was up-regulated significantly (P<0.01), and that of hippocampal Bax protein downregulated considerably in the EA group (P<0.01). No significant differences were found between AD model and ONT + EA groups in the expression of hippocampal Bcl-2 and Bax proteins (P>0.05), suggesting a necessary premise condition for EA in regulating hippocampal Bcl-2 and Bax expression. CONCLUSION: "Xiusanzhen" can regulate the expression of hippocampal Bcl-2 and Bax proteins in AD rats, which may contribute to its clinical effect in relieving AD, and the therapeutic effect depends on the integrity of the olfactory nerve pathway.

[Micro-acupuncture system in the hairline region of the scalp].

Studying on the acupoints in the hairline region of the scalp, it is discovered that a group of acupoints are orderly arranged in the hairline region of the scalp. Furthermore, the arrangement is highly consistent with the meridian route of zang-fu organs. Manipulating needles at acupoints in the hairline regions of the scalp may treat certain diseases, including insomnia, migraine, and Alzheimer disease. The characteristics of the acupuncture treatment at acupoints in the hairline of scalp resemble the characteristics of ear acupuncture and eye acupuncture. Thus forms a special micro-acupuncture system in the hairline regions of the scalp. This paper states the theoretical basis, the arrangement of acupoints, clinical application and operating norms.

Study on perfume stimulating olfaction with volatile oil of Acorus gramineus for treatment of the Alzheimer's disease rat.

OBJECTIVE: To probe into the therapeutic effect of perfume stimulating olfaction with volatile oil of Acorus Gramineus on the Alzheimer's disease (AD) rat. METHODS: Totally 50 adult SD rats, male,weighing 300 +/- 10 g, were randomly divided into 5 groups, normal group (group A), olfactory nerve severing model group (group B), AD model group (group C), AD model plus perfume stimulation group (group D), AD model olfactory nerve severing plus perfume stimulation group (group E), 10 rats in each group. After perfume stimulation, Morris maze test was conducted for valuating the learning and memory ability; Malondaldehyde (MDA) content, and superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities in the brain, and the brain weight were detected. RESULTS: Compared with the AD model group, the average escape latency and swimming distance in 6 days were significantly shorter than those in the group A, B, D (P < 0.01), with no significant differences between the group C and the group E (P > 0.05); Compared with the group A, B and D, MDA content in the group C significantly increased (P < 0.01), and SOD and GSH-Px activities significantly decreased (P < 0.01), and brain weight/body weight decreased significantly in the group C (P < 0.01), with no significant differences between the group C and the group E (P > 0.05). CONCLUSION: Perfume stimultating olfaction with volatile oil of Acorus Gramineus can significantly increase the learning-memory ability, decrease MDA content and increase SOD and GSH-Px activities and weight of brain in AD rats.

[Effect of electroacupuncture on cerebral cortex ultrastructure in rats with cerebral ischemia-reperfusion injury].

OBJECTIVE: To observe the effect of electroacupuncture (EA) of "Baihui" (GV 20) and "Dazhui" (GV 14) on the ultrastructure of cerebral cortex in rats with cerebral ischemia reperfusion (CI/R) injury. METHODS: Fifteen SD rats were equally randomized into sham-operation (sham), model and EA groups. CI/R model was established by right middle cerebral artery occlusion for 1 h and reperfusion for 24 h. EA (20 Hz/80 Hz, 1-3 mA) was applied to "Baihui" (GV 20) and "Dazhui" (GV 14) for 30 min. The ischemic cerebral cortex tissue was taken, fixed in 2.5% glutaral solution, embedded in araldite, cut into sections and stained for observing the ultrastructure changes of cortical pyramidal cells, astrocyte and blood brain barrier (BBB) under transmission electronic microscope. RESULTS: In sham group, the anatomical structure of the cerebral pyramidal cells was normal basically. In model group, the neuronal mitochondria and the capillary endothelium and the processes of the astrocyte got swelling, the mitochondrial cristae were broken, the capillary lumens became narrow or were blocked up. In EA group, the injury degree of the pyramidal cells, glial cells and BBB were lighter. CONCLUSION: EA can reduce ischemic injury of the cerebral cortical neurons and blood brain barrier in rats with CI/R injury.

[Effect of "Xiusanzhen" on learning-memory ability and hippocampal somatostatin and arginine vesopressin contents in vascular dementia rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) of "Xiusanzhen" [bilateral "Yingxiang" (LI 20) + "Yintang" (GV 29)] on learning-memory ability and hippocampal somatostatin (SS) and arginine vasopressin (AVP) contents in vascular dementia (VD) rats. METHODS: Forty SD rats were randomly divided into control, VD model, VD plus olfactory bulb lesion (OBL, destroyed by electro-coagulation) and EA groups, with 10 cases in each. VD model was established by 4-vessel occlusion. Morris maze tests were conducted for evaluating the rats' learning and memory ability. EA (1-3 mA, 80/100 Hz) was applied to bilateral LI 20 and GV 29 in VD + OBL and EA groups for 10 min, once daily (except Saturdays and Sundays) for 6 weeks. The contents of SS and AVP in hippocampus tissue were measured by radioimmunoassay. RESULTS: In comparison with control group, the average escape latency (AEL) prolonged significantly and the target-platform crossing times (TPCT) decreased remarkably in VD model group (P < 0.01). In comparison with VD model group, the AEL was shortened and TPCT increased remarkably in EA group (P < 0.01). No significant differences were found between VD model and VD + OBL groups in the AEL and the TPCT (P > 0.05). Hippocampal SS and AVP contents in VD model group were significantly lower than those in control group (P < 0.05), while those in EA group were markedly higher than those in VD model group (P < 0.05). No significant differences were found between VD model and VD + OBL groups in SS and AVP contents (P > 0.05). CONCLUSION: EA of "Xiusanzhen" can improve VD rats' learning-memory ability, which may be related to its effects in raising hippocampal SS and AVP contents and to the intact olfactory pathway.

[Effect of "Xiusanzhen" on learning-memory ability and hippocampal ChAT and AChE activity in Alzheimer disease rats].

OBJECTIVE: To explore the effect of "Xiusanzhen" [bilateral "Yingxiang" (LI 20) + "Yintang" (EX-HN 3)] on learning-memory ability and choline acetyl transferase (ChAT) and acetyl cholinesterase (AChE) activity of hippocampus tissue in Alzheimer Disease (AD) rats. METHODS: A total of 40 Sprague Dawley rats were randomly divided into control, AD model, AD + olfactory-bulb lesion (OBL, induced by electrocoagulation method) + EA and "Xiusanzhen" (AD + EA) groups. AD model was established by 4-vessel occlusion. Morris water maze tests were conducted for evaluating the rat's learning-memory ability. The activity of ChAT and AChE of hippocampus tissue was detected by colorimetric method. RESULTS: In comparison with control group, the average escape latency (AEL) prolonged significantly and the target-platform crossing times decreased remarkably in AD model group (P<0.01). In comparison with AD model group, the AEL shortened and the target-platform crossing times increased remarkably in AD + EA group (P<0.01). No significant differences were found between AD model and AD + OBL + EA groups in AEL and the target-platform crossing times (P>0.05). The ChAT and AChE activity of hippocampus in AD model group was significantly lower than that of control group (P<0.05), while that of ChAT and AChE in AD + EA group was significantly higher than that in AD model group (P<0.05). No significant difference was found between AD model and AD + OBL + EA groups in ChAT and AChE activity of hippocampus (P>0.05). CONCLUSION: EA of "Xiusanzhen" can markedly raise the learning-memory ability in AD rats, which may be related to its effect in increasing the ChAT and AChE activity of hippocampus in AD rats. Its therapeutical effect also depends on the intact of the olfactory pathway.

[Involvement of melatonin in the adjusting effect of electroacupuncture in resisting oxygen stress in cerebral ischemia-reperfusion injury rats].

OBJECTIVE: To explore the effect of electroacupuncture (EA) in resisting acute cerebral ischemia-reperfusion injury (CI-RI) via anti-oxidation of melatonin (MT). METHODS: A total of 52 male SD rats were randomized into Sham-operation (sham, n=8), CI-RI (model, n=8), EA (n=8), MT (3.2 mg/Kg, i.p., n=8), EA+MT (n=8), EA+Luzindole (L, a blocker of MT, 1 mg/100 microl/rat, n=6), EA+PD 98059 (a blocker of mitogen-activated protein kinase, MAPK, 1 mg 100 microl/rat, n=6) groups. EA (3 Hz,1-3 mA, continuous waves) was applied to "Dazhui" (GV 14) and "Baihui" (GV 20) for 30 min. CI-RI model was induced by right middle cerebral artery occlusion (MCAO) for 60 min and reperfusion for 24 h except sham-operation group. MT content of pineal gland tissue was determined by high performance liquid chromatography (HPLC) and neuroethological scores of the rats were evaluated by Kuluz's and Julio's methods. Bax and Bcl-2 gene protein expression of the striate body was detected by immunohistochemistry. RESULTS: In comparison with sham group, the number of Bax immunoreactive (IR) positive cells and Bax/Bcl-2 in striate body increased significantly (P<0.001), and neuroethological score and Bcl-2 IR-positive cells decreased considerably in model group (P<0.01). Compared with model group, pineal MT content, neuroethological score and Bcl-2 IR-positive cells in EA group, neuroethological scores in EA+L and EA+P groups, and Bcl-2 expression in MT and EA+MT groups increased considerably (P<0.01, 0.001); while Bax IR-positive cells and Bax/Bcl-2 in EA, MT and EA+MT groups decreased very significantly (P<0.001). It showed that EA could obviously increase pineal MT content, improve CI-RI rats' activity score, upregulate Bcl-2 expression and down-regulate Bax expression in striate body. In comparison with simple EA group, neuroethological scores of EA+L and EA+P groups, Bax IR-positive cells and Bax/Bcl-2 of EA+MT group were significant lower (P<0.05, 0.01). No significant differences were found between EA+L and EA+P groups in behavior scores, and between MT and EA+MT groups in Bax and Bcl-2 IR-positive cell number and Bax/Bcl-2 (P>0.05). CONCLUSION: EA has a good effect of anti-oxygen stress, protecting the brain from ischemic damage directly or indirectly, which may be related to its effect in upregulating the synthesis and release of MT.

[Clinical investigation on electroacupuncture treatment of vascular dementia with "Xiusanzhen"].

OBJECTIVE: To observe the effect of electroacupuncture (EA) in the treatment of vascular dementia (VD) so as to explore an effective therapy for it. METHODS: Sixty cases of VD were randomly divided into acupuncture group and medication (Duxil, 40 mg/time, 2 times/d for 10 weeks) group, with 30 cases in each group. "Xiusanzhen" means that two acupuncture needles were penetrated subcutaneously from bilateral Yingxiang (LI 20) to Shangyingxiang (EX-HN 8) respectively, and the 3rd needle was penetrated from the midpoint between two Yangbai (GB 14) through Yintang (EX-HN 3) to the nasal root, followed by applying EA stimulation to the acupoints. Hasegawa's Dementia Scale (HDS), Mini-mental state examination (MMSE), and Functional Activities Questionnaire (FAQ) were used for assessment of the VD patient's state before and after the treatment. RESULTS: After the treatment, the total effective rates of acupuncture and medication groups were 80.00% and 73.33% respectively. The effective rate of acupuncture group was significantly superior to that of medication group (P<0.05). Before treatment, scores of HDS, MMSE and FAQ in both groups had no significant differences (P>0.05); after the treatment, the scores of HDS and MMSE increased significantly, and that of FAQ decreased remarkably in both groups (P<0.01), and the difference of scores (or absolute value of FAQ) between post-treatment and pre-treatment in EA group was obviously higher than that in medication group (P<0.05). CONCLUSION: Acupuncture of "Xiusanzhen" is effective in the treatment of VD patients.

[Study on the morphological evidence of five-shu-point].

OBJECTIVE: To search for the morphological evidence of five-shu-point so as to explain the theory of the five-shu-point more reasonably. METHODS: Carefully study on Yellow Emperor's Internal Classic and compare with the modern anatomic knowledge. CONCLUSION: The theory of five-shu-point is closely related with the theory of blood vessels. The distributional pattern of the superficial vein on the remote parts of the limbs (below the elbow or the knee joints) is the morphological basis for the theory of five-shu-point.

[Observation on therapeutic effect of puncturing with needles in alignment at the hairline region on vascular dementia].

OBJECTIVE: To search for an effective therapy for vascular dementia (VD). METHODS: One hundred and twenty cases of VD were randomly divided into 2 groups, 60 cases in each group. The observation group were treated with puncturing with needles in alignment at the hairline region and the control group with oral administration of Duxil. Hasegawa's dementia scale (HDS), mini-mental state examination (MMSE), functional activity questionnaire (FAQ), nerve function defect (NFD) and main symptoms (MS) scales were used for assessment of symptoms and signs before and after treatment. RESULTS: Changes of scores for HDS, MMSE, FAQ, NFD and MS before and after treatment had statistically very significant differences (P < 0.01) in the two groups, with significant difference between the two groups (P < 0.05). The total effective rate was 88.33% in the observation group and 81.67% in the control group with a significant difference between the two groups (P < 0.05). CONCLUSION: Puncturing with needles in alignment at the hairline region has definite therapeutic effect on vascular dementia.

[Protective effect of electroacupuncture of "xiusanzhen" on neuronal mitochondria in rats with acute cerebral ischemia-reperfusion injury].

OBJECTIVE: To explore the protective effect of electroacupuncture (EA) of "Xiusanzhen"([Chinese characters: see text]) on mitochondria of cortical neurons in acute cerebral ischemia-reperfusion (CI/R) rats. METHODS: A total of 30 cases of SD rats were divided into control, model and EA groups. CI/R model was established by occlusion of the middle cerebral artery and reperfusion. EA (80-100 Hz, 1-3 mA) was applied to "Xiusanzhen" (the median line of the nose root, 2 mm to the lateral median line on the bilateral sides) for 60 min. Content of serum reduced glutathione (GSH) and glutathion peroxidase (GSH-Px) activity were detected by using Tietze's reducase assay and dithio-bis-nitrobenzoic acid (DTNB) direct method respectively. The surface density (Sv), numerical density (Nv) and perimeter/area (Pv) of the mitochondria of the cerebral neurons were detected using stereological technique and electronic microscope. RESULTS: Compared with control group, Sv and Pv of model group increased significantly (P < 0.05), while serum GSH content and GSH-Px activity of model group decreased considerably (P < 0.05). While in comparison with model group, Sv and Pv of EA group lowered obviously (P < 0.05), and GSH and GSH-Px both increased pronouncedly (P < 0.05). No significant difference was found among 3 groups in Nv (P > 0.05). CONCLUSION: EA of "Xiusanzhen" has a definite protective effect on mitochondria of cortical neurons in acute focal cerebral ischemia-reperfusion rats, which may be related with its antioxidation.