Homocysteine-Induced Disturbances in DNA Methylation Contribute to Development of Stress-Associated Cognitive Decline in Rats.

Chronic stress is generally accepted as the main risk factor in the development of cognitive decline; however, the underlying mechanisms remain unclear. Previous data have demonstrated that the levels of homocysteine (Hcy) are significantly elevated in the plasma of stressed animals, which suggests that Hcy is associated with stress and cognitive decline. To test this hypothesis, we analyzed the cognitive function, plasma concentrations of Hcy, and brain-derived neurotropic factor (BDNF) levels in rats undergoing chronic unpredicted mild stress (CUMS). The results showed that decreased cognitive behavioral performance and decreased BDNF transcription and protein expression were correlated with hyperhomocysteinemia (HHcy) levels in stressed rats. Diet-induced HHcy mimicked the cognitive decline and BDNF downregulation in the same manner as CUMS, while Hcy reduction (by means of vitamin B complex supplements) alleviated the cognitive deficits and BDNF reduction in CUMS rats. Furthermore, we also found that both stress and HHcy disturbed the DNA methylation process in the brain and induced DNA hypermethylation in the BDNF promoter. In contrast, control of Hcy blocked BDNF promoter methylation and upregulated BDNF levels in the brain. These results imply the possibility of a causal role of Hcy in stress-induced cognitive decline. We also used ten-eleven translocation (TET1), an enzyme that induces DNA demethylation, to verify the involvement of Hcy and DNA methylation in the regulation of BDNF expression and the development of stress-related cognitive decline. The data showed that TET1-expressing viral injection into the hippocampus inhibited BDNF promoter methylation and significantly mitigated the cognitive decline in HHcy rats. Taken together, novel evidence from the present study suggests that Hcy is likely involved in chronic stress-induced BDNF reduction and related cognitive deficits. In addition, the negative side-effects of HHcy may be associated with Hcy-induced DNA hypermethylation in the BDNF promoter. The results also suggest the possibility of Hcy as a target for therapy and the potential value of vitamin B intake in preventing stress-induced cognitive decline.

Perilla Leaf Extract Attenuates Asthma Airway Inflammation by Blocking the Syk Pathway.

Perilla frutescens (L.) Britton is a classic herbal plant used widely against asthma in China. But its mechanism of beneficial effect remains undermined. In the study, the antiallergic asthma effects of Perilla leaf extract (PLE) were investigated, and the underlying mechanism was also explored. Results showed that PLE treatment significantly attenuated airway inflammation in OVA-induced asthma mice, by ameliorating lung pathological changes, inhibiting recruitment of inflammatory cells in lung tissues and bronchoalveolar lavage fluid (BALF), decreasing the production of inflammatory cytokines in the BALF, and reducing the level of immunoglobulin in serum. PLE treatment suppressed inflammatory response in antigen-induced rat basophilic leukemia 2H3 (RBL-2H3) cells as well as in OVA-induced human peripheral blood mononuclear cells (PBMCs). Furthermore, PLE markedly inhibited the expression and phosphorylation of Syk, NF-κB, PKC, and cPLA2 both in vivo and in vitro. By cotreating with inhibitors (BAY61-3606, Rottlerin, BAY11-7082, and arachidonyl trifluoromethyl ketone) in vitro, results revealed that PLE's antiallergic inflammatory effects were associated with the inhibition of Syk and its downstream signals NF-κB, PKC, and cPLA2. Collectively, the present results suggested that PLE could attenuate allergic inflammation, and its mechanism might be partly mediated through inhibiting the Syk pathway.

Mallotus oblongifolius extracts ameliorate ischemic nerve damage by increasing endogenous neural stem cell proliferation through the Wnt/ß-catenin signaling pathway.

Mallotus oblongifolius (MO), an edible medicinal plant from Hainan in China, shows a wide range of bioactivities. The daily consumption of MO or its extracts has been observed to ameliorate ischemic nerve injury. However the mechanisms remain unclear. In this study, the effects of MO both in vitro and in vivo were investigated. The results indicated that MO improved the motor ability, neurosensory ability, balance and grasping ability of mice with ischemic injuries, induced by bilateral common carotid artery ligation (BCCAL). In addition, MO improved the morphology of neurons, resisted the loss of neurons, and enhanced the content of the nestin protein in the cerebral cortex and subgranular zone (SGZ) area. Furthermore, in the oxygen-glucose deprivation and reperfusion (OGD/R) treated cell model, MO could effectively activate the Wnt/ß-catenin signaling pathway and promote the proliferation of neural stem cells (NSCs) and increase the protein expression levels of ß-catenin and CyclinD1. Our results suggest that Mallotus oblongifolius may be used as nutraceuticals or functional foods to alleviate ischemic nerve damage and promote recovery from ischemic stroke.

[The effect of baicalin on human brain microvascular endothelial cell under the glucose deprivation combined with hypoxia condition].

OBJECTIVE: To investigate the mechanisms of baicalin on anti-cerebral ischemic through observing the effect of baicalin on human brain microvascular endothelial cell under the glucose deprivation combined with hypoxia condition. METHODS: Human brain microvascular endothelial cells (HBMVECs) cultured in vitro were divided into the following groups: normal group, model group, baicalin high dose group, baicalin middle dose group, baicalin low dose group, nimodipine group. The kits were used to detect the cell viability, leakage rate of lactate dehydrogenase (LDH), Na(+)-K(+)-ATPase, Ca(2+)-ATPase, the concentration of Ca2+ in each group, and apoptosis rates of each group were analyzed by flow cytometry (FCM). RESULTS: Compared with the normal group, the cell viability, Na(+)-K(+)-ATPase and Ca(2+)-ATPase in model group decreased significantly (P < 0.01, P < 0.05). But the leakage rate of LDH, Ca2+ in cells and apoptosis rates increased remarkably (P < 0.01). Compared with the model group, the cell viability, Na(+)-K(+)-ATPase and Ca(2+)-ATPase increased obviously (P < 0.01, P < 0.05) in baicalin high dose group. But the leakage rate of LDH and Ca2+ in cells in baicalin high dose group decreased significantly comparing with that of model group (P < 0.05, P < 0.01). And the reduction of intracellular Ca2+ was superior to that of nimodipine group. Meanwhile, the apoptosis rates decreased significantly in both baicalin high and middle dose groups (P < 0.01). CONCLUSION: Baicalin could improve the cell viability of HBMVECs under the glucose deprivation combined with hypoxia condition. And the mechanisms were related with improving the energy metabolism, inhibiting intracellular calcium overload and decreasing the apoptosis rate of cells further.

[Effect of acupuncture therapy on appetite of obesity patients].

OBJECTIVE: To observe the effect of acupuncture intervention on the appetite of obesity patients. METHODS: A total of 118 obesity patients were randomized into acupuncture group (76 cases, treated by true acupuncture needles) and placebo group (42 cases, treated by placebo acupuncture needles) using single-blind method. All the patients of the two groups were ordered to control their diet during the treatment. The acupoints around the umbilicus [Zhongwan (CV 12), Zhongji (CV 3), Daheng (SP 15), Xiawan (CV 10), Shimen (CV 5) and Tianshu (ST 25), etc.] and Liangqiu (ST 34), Zusanli (ST 36), and Yin-lingquan (SP 9) were punctured with filiform needles which were manipulated with uniform reducing and reinforcing method for a while tijl "Deqi" and retained for 30 min. The treatment was conducted once every other day, 12 times altogether. Body mass index (BMI), and visual analogue scale (VAS) scores of eating-desire and hunger feeling and prospective food consumption were measured before and after the treatment. The gastric fluid survival rate (GFSR) was evaluated by using ultrasound scanning. RESULTS: The BMI in the acupuncture group was obviously declined after the treatment in comparison with the placebo group (P < 0.01). Compared to the placebo group, the VAS scores of eating-desire, hunger feeling and prospective food consumption were significantly decreased in the acupuncture group ( P < 0.05), but there are no significant difference between two groups in the VAS score of gastric fullness feeling (P > 0.05). The GFSR was obviously increased in the acupuncture group compared to the placebo group (P < 0.05). CONCLUSION: Acupuncture therapy can significantly decrease BMI and delay the digesting time and control the appetite in obesity patients, which may contribute to its effect in body weight reduction.

[Clinical observation on simple obesity treated by acupuncture].

OBJECTIVE: To evaluate the effectiveness and safety of simple obesity treated by acupuncture. METHODS: By randomized single-blind clinical trial, one hundred and eighteen cases of simple obesity were divided into an acupuncture group (76 cases) and a placebo-acupuncture control group (42 cases), additionally, health control group (30 cases) was included. In acupuncture group and placebo-acupuncture control group, all the patients received a restricted diet; Zhongwan (CV 12) and Zhongji (CV 3) etc. at abdomen and Liangqiu (ST 34) and Zusanli (ST 36) etc. at limbs were selected; body mass index (BMI), Serum Total Cholesterol, triglyceride (TG), Glucose, Creatinine, urea nitrogen (BUN), Uric Acid and adverse reactions scores were observed. RESULTS: After treatment the BMI in acupuncture grown was lower than that in placebo-acupuncture control group (P < 0.01). In metabolism indices, the serum Total Cholesterol and Glucose after treatment were reduced obviously than those before treatment in acupuncture group (all P < 0.01), and there was no significant differences in other metabolism indices (all P > 0.05) in two groups. After treatment, in adverse reactions scores, the hunger sensation scores in acupuncture group was reduced than that in placebo-acupuncture control group (P < 0.05), and there was no significant differences in other indices (all P > 0.05). CONCLUSION: BMI of simple obesity was reduced by acupuncture, and the Serum Total Cholesterol and Glucose were reduced accordingly. The adverse reac tions such as weakness, nervosa and diarrhea, etc. doesn't appear after acupuncture treatment. Acupuncture therapy is one of the safe and effective methods for simple obesity.

[Effect of acupoint application therapy on expression of IL-4 and IFN-gamma in lung tissue of asthma rats].

OBJECTIVE: To observe the effect of acupoint application of Chinese herbal medicines on pathological changes of pulmonary tissue in asthma rats so as to explore its underlying mechanism. METHODS: A total of 33 Wistar rats were randomized into normal control (n=6), model (n=9), dexamethasone (DX, n=9) and acupoint-application (A-A, n=9) groups. Asthma model was established by intraperitoneal injection of 10% ovalbumin(OVA, 1 mL)and forced inhalation of atomized 2% OVA (25 mL) for 40 min, once daily for 2 weeks. "Dazhui" (GV 14). "Pishu" (BL 20), "Feishu" (BL 13) and "Shenshu" (BL 23) were selected for external application of Chinese medicinal herbs (pricklyash peel, white mustard seeds, asarum herb, etc.) for rats of A-A group, and that of control drugs (ang-kak, black rice, ginger juice, etc) for rats of DX group. Infiltration degrees of eosinophils (Eos), lymphocytes (L) and macrophages (MO) in the pulmonary tissue were observed under microscope. IL-4 and IFN-gamma expression was displayed by using immunohistochemical method. RESULTS: In comparison with control group, the infiltration degrees of E(OS). L and Mphi increased significantly in model, DX and acupoint-application groups (P < 0.05, 0.01); while compared with model group, the infiltration of E(OS), L and Mphi in DX group, E(OS) in A-A group decreased significantly (P < 0.05, 0.01). No significant differences were found between DX and A-A groups in these 3 indexes (P > 0.05). Results of immunohisto chemical staining in the lung tissue indicated that compared with control group, IL-4 expression in model increased significantly (P < 0.01), IFN-gamma expression decreased considerably in model and DX group (P < 0.01); when compared with model group, IL-4 in both A-A group and DX group decreased significantly, and IFN-gamma expression in A-A group increased considerably (P < 0.05, 0.01). No significant differences were found between DX and A-A groups in IL-4 expression (P > 0.05). CONCLUSION: Acupoint application of Chinese herbal medicines can balance Thl/Th2 and effectively reduce the infiltration of eosinophils, which may contribute to its therapeutic effect in relieving asthma.