Electroacupuncture Alleviates Neuroinflammation by Regulating Microglia Polarization via STAT6/PPARγ in Ischemic Stroke Rats.

Previous study showed that electroacupuncture (EA) produced a protective effect on cerebral ischemia-reperfusion injury (CIRI) in rats and may correlate with the anti-inflammatory effects of microglia. This study aimed to investigate further whether EA could modulate neuroinflammation by targeting the Signal Transducer and Activator of Transcription 6 (STAT6) and Peroxisome Proliferator-Activated Receptor γ (PPARγ) pathway, the key regulator of microglia. Middle cerebral artery occlusion (MCAO) rats were used, and 6 h after reperfusion, EA interventions were performed in Chize (LU 5), Hegu (LI 4), Sanyinjiao (SP 6), and Zusanli (ST 36) on the affected side of the rats, the group that received EA + STAT6 phosphorylation inhibitor AS1517499 was used as a parallel control. The degree of neurological impairment, infarct volume, microglia polarization, inflammation levels and activity of STAT6/PPARγ pathway were then assessed by neurological deficit score, triphenyl tetrazolium chloride (TTC) staining, immunofluorescence, western blotting (WB), quantitative real-time PCR (qPCR) and Enzyme linked immunosorbent assay (ELISA). The data showed that EA significantly alleviated nerve injury, reduced infarct volume, enhanced the expression and activity of STAT6/PPARγ pathway, inhibited NF-κB activity, increased M2 microglia numbers and anti-inflammatory factor release, and inhibited microglia M1-type polarization and pro-inflammatory factor expression. In contrast, inhibition of STAT6 phosphorylation exacerbated neural damage, inhibited STAT6/PPARγ pathway activity, promoted microglia M1-type polarization and exacerbated neuroinflammation, resulting in an attenuated positive effect of EA intervention. Therefore, we concluded that EA intervention could attenuate microglia-associated neuroinflammation by enhancing the expression and activity of STAT6/PPARγ pathway, thereby reducing CIRI in MCAO rats.

A Review of Neuroprotective Effects and Mechanisms of Ginsenosides From Panax Ginseng in Treating Ischemic Stroke.

Ischemic stroke has been considered one of the leading causes of mortality and disability worldwide, associated with a series of complex pathophysiological processes. However, effective therapeutic methods for ischemic stroke are still limited. Panax ginseng, a valuable traditional Chinese medicine, has been long used in eastern countries for various diseases. Ginsenosides, the main active ingredient of Panax ginseng, has demonstrated neuroprotective effects on ischemic stroke injury during the last decade. In this article, we summarized the pathophysiology of ischemic stroke and reviewed the literature on ginsenosides studies in preclinical and clinical ischemic stroke. Available findings showed that both major ginsenosides and minor ginsenosides (such as Rg3, Rg5, and Rh2) has a potential neuroprotective effect, mainly through attenuating the excitotoxicity, Ca2+ overload, mitochondria dysfunction, blood-brain barrier (BBB) permeability, anti-inflammation, anti-oxidative, anti-apoptosis, anti-pyroptosis, anti-autophagy, improving angiogenesis, and neurogenesis. Therefore, this review brings a current understanding of the mechanisms of ginsenosides in the treatment of ischemic stroke. Further studies, especially in clinical trials, will be important to confirm the clinical value of ginseng and ginsenosides.

Mechanism of Electroacupuncture Against Cerebral Ischemia-Reperfusion Injury: Reducing Inflammatory Response and Cell Pyroptosis by Inhibiting NLRP3 and Caspase-1.

Cell pyroptosis is one of the main forms of neuronal injury after cerebral ischemia-reperfusion. It is accompanied by an inflammatory reaction and regulated by the caspase gene family. Electroacupuncture (EA) can reduce neuronal injury caused by cerebral ischemia-reperfusion, and we speculated that EA can prevent neuronal pyroptosis after cerebral ischemia-reperfusion by regulating the nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3)/caspase-1 pathway. The cerebral ischemia-reperfusion injury model of C57 and caspase-1 gene knockout (Cas-1 ko) mice was established by Longa's method. EA was conducted at acupoints Chize (LU5), Hegu (LI4), Sanyinjiao (SP6), and Zusanli (ST36) for 1.5 h after cerebral ischemia-reperfusion injury for 20 min, and observation was carried out after 24 h. Neurological deficit scores evaluated the neurological function, cerebral infarction volume was observed by triphenyl tetrazolium chloride (TTC) staining, hematoxylin and eosin (H&E) staining, TUNEL and caspase-1 double-labeled fluorescence staining, and NLRP3 and caspase-1 double-labeled immunofluorescence staining that were used to observe the morphology of neurons in hippocampus, and the protein expression of NLRP3, pro-caspase-1, cleaved caspase-1 p20, pro-interleukin-1ß (IL-1ß), cleaved IL-1ß, and GSDMD was detected by Western blot assay. Results showed that EA could reduce the score of neurological deficit, reduce the volume of cerebral infarction and improve the degree of nerve cell injury, and inhibit NLRP3, pro-caspase-1, cleaved caspase-1 p20, pro-IL-1ß, cleaved IL-1ß, and GSDMD protein expression. In summary, EA plays a neuroprotective role by reducing the pyroptotic neurons that were caspase 1-mediated and inflammatory response after cerebral ischemia-reperfusion.

Dopaminergic signaling in prefrontal cortex contributes to the antidepressant effect of electroacupuncture: An iTRAQ-based proteomics analysis in a rat model of CUMS.

Electroacupuncture (EA) is used as an adjunctive treatment for depression. This study was conducted to evaluate the efficacy and mechanisms of EA in the depressive rat model induced by chronic unpredictable mild stress (CUMS) in male adult Wistar rats. The underlying mechanisms were explored by using isobaric tags for relative and absolute quantitation (iTRAQ)-based proteomic analysis of the proteins in the prefrontal cortex (PFC), and observing the number of the PFC neurons stained with hematoxylin and eosin (H&E) and synaptic morphological changes under transmission electron microscopy (TEM). The results showed that EA plus paroxetine (EA + Par) for 1 week significantly relieved depression-like anhedonia symptoms and improved anxiety-like behavior, accompanied by the improvements in synaptic morphology and a significant increase of PFC neurons. Moreover, EA or paroxetine alone significantly alleviated anhedonia symptoms after 2 weeks of intervention. Additionally, iTRAQ analysis showed that dopaminergic signaling was significantly altered in CUMS rats after 1 week of EA treatment. As the critical enzyme of this pathway, aromatic-l-amino-acid decarboxylase (DDC) was significantly upregulated after the treatment with EA + Par for 1 week. These findings suggested that the dopaminergic signaling pathway in PFC may be involved in the antidepressant mechanisms of EA.

Electroacupuncture alleviated the depression-like behavior by regulating FGF2 and astrocytes in the hippocampus of rats with chronic unpredictable mild stress.

Studies have shown that basic fibroblast growth factor (FGF2) is a neurotrophic factor associated with depression. Electroacupuncture (EA) has been shown to be an effective treatment for depression. In the current study, we observed the effects of EA on hippocampal FGF2 and astrocytes, and further investigated the mechanism underlying antidepressant effect of EA. The chronic unpredictable mild stress (CUMS) method were selected to induce depressive-like behaviors of rats. Paroxetine is a commonly used antidepressant and was used as a positive control drug in this experiment. The male adult Sprague Dawley (SD) rats were randomized to four experimental groups (normal control group, CUMS group, EA group and paroxetine group, n = 10/group). EA intervention was administered once daily for 14 days at acupuncture points Baihui (GV20) and Yintang (GV29). Rats in the paroxetine group received daily paroxetine administered intragastrical. Behavioral test, immunohistochemistry (IHC), western blot (WB) and quantitative real-time PCR (qPCR) were conducted to evaluate the intervene effect and the changes of FGF2 and astrocyte marker (glial fibrillary acidic protein, GFAP). The results showed that EA and paroxetine could improve depression-like behavior in CUMS rats, and up-regulated the expression level of FGF2 in the hippocampus, increased GFAP protein expression and the mean optical density of GFAP-immunoreactive astrocyte (GFAP-ir astrocyte). Our findings have identified that EA could ameliorate depressive-like behaviors possibly by regulating the expression of FGF2 in the hippocampus, and the mechanism might be related to the effect of FGF2 on astrocytes.

EA Ameliorated Depressive Behaviors in CUMS Rats and Was Related to Its Suppressing Autophagy in the Hippocampus.

Autophagy is confirmed to be involved in the onset and development of depression, and some antidepressants took effect by influencing the autophagic process. Electroacupuncture (EA), as a common complementary treatment for depression, may share the mechanism of influencing autophagy in the hippocampus like antidepressants. To investigate that, sixty Sprague-Dawley rats firstly went through chronic unpredictable mild stress (CUMS) model establishment, and 15 rats were assigned to a control group. After modeling, 45 successfully CUMS-induced rats were randomly divided to 3 groups: CUMS, selective serotonin reuptake inhibitor (SSRI), and EA groups (15 rats per group), to accept different interventions for 2 weeks. A sucrose preference test (SPT), weighing, and open field test (OFT) were measurement for depressive behaviors of rats. Transmission electron microscope (TEM), immunohistochemistry (IHC), and western blot analysis were used to evaluate the autophagic changes. After that, depression-like behaviors were successfully induced in CUMS models and reversed by SSRI and EA treatments (both p < 0.05), but these two therapies had nonsignificant difference between each other (p > 0.05). Autolysosomes observed through TEM in the CUMS group were more than that in the control group. Their number and size in the SSRI and EA groups also decreased significantly. From IHC, the CUMS group showed enhanced positive expression of both Beclin1 and LC3 in CA1 after modeling (p < 0.05), and the LC3 level declined after EA treatments, which was verified by decreased LC3-II/LC3-I in western blot analysis. We speculated that CUMS-induced depression-like behavior was interacted with an autophagy process in the hippocampus, and EA demonstrated antidepressant effects by partly inhibiting autophagy with a decreased number of autolysosomes and level of LC3 along with LC3-II/LC3-I.

Electroacupuncture improves synaptic plasticity by regulating the 5-HT1A receptor in hippocampus of rats with chronic unpredictable mild stress.

OBJECTIVES: To investigate the antidepressant effects of electroacupuncture (EA) on chronic unpredictable mild stress (CUMS) in rats, as well as the effects of EA on hippocampal neurons, synaptic morphology, and 5-hydroxytryptamine (HT) receptor expression. METHODS: Forty adult male Wistar rats were randomly divided into normal control, CUMS, EA, and paroxetine groups. CUMS modeling was performed for 21 days, followed by 14 days of intervention: rats in the EA group underwent stimulation of GV20 and GV29 acupuncture points for 30 minutes daily; rats in the paroxetine group were administered paroxetine daily. Behavioral tests, transmission electron microscopy, western blotting, and real-time quantitative polymerase chain reaction were used to evaluate the effects of the intervention. RESULTS: EA treatment reversed the behavioral changes observed in rats due to CUMS modeling; it also improved the pathological changes in organelles and synaptic structures of hippocampal neurons, and upregulated the protein and mRNA expression levels of 5-HT1A receptor. There were no significant differences in 5-HT1B receptor protein and mRNA expression levels among the groups. CONCLUSIONS: EA treatment can alleviate depression-like symptoms in CUMS rats. The underlying mechanism may include promoting the expression of 5-HT1A receptor mRNA and protein, thereby improving synaptic plasticity in the hippocampus.

Four new chemical constituents from Piper pleiocarpum.

Two new polyoxygenated cyclohexenes (1-2), one new benzoate derivative (3), and one new dineolignan (4) together with one known neolignan (5) were isolated from whole plants of Piper pleiocarpum. The structures of these compounds were determined by extensive spectroscopic methods including 1D, 2D NMR, HR-ESI-MS, and by comparison with the literature. The 13C NMR spectra of the known compound 5 were completely assigned for the first time. All isolated compounds (1-5) were evaluated for their cytotoxic activities against five human cancer cell lines (including A-549, SMMC-7721, HL-60, MCF-7, and SW-480), Only compound 4 showed inhibitory activity against MCF-7 cell line with IC50 value of 18.24 ± 0.69 µM.

iTRAQ-Based Protein Profiling in CUMS Rats Provides Insights into Hippocampal Ribosome Lesion and Ras Protein Changes Underlying Synaptic Plasticity in Depression.

Hippocampal atrophy is one of the key changes in the brain implicated in the biology of depression. However, the precise molecular mechanism remains poorly understood due to a lack of biomarkers. In this research, we used behavioral experiments to evaluate anxiety and anhedonia levels in depressed rats using chronic unpredictable mild stress (CUMS) modeling. We also used isobaric tag for relative and absolute quantitation (iTRAQ) to identify the differentially expressed hippocampal proteins between depressed and normal rats. Bioinformatics analyses were also performed for a better understanding. The results showed that CUMS rats had higher anxiety and anhedonia levels than control rats, along with hippocampal lesions. Through iTRAQ and bioinformatics analyses, we found that ribosome proteins were significantly downregulated and Ras proteins exhibited a mixed change in the hippocampus of depressed rats. These findings suggest that the expression of hippocampal ribosome lesions and Ras proteins is significantly different in depressed rats than in control rats, providing new insights into the neurobiology of depression.

Electroacupuncture Improves Antidepressant Effects in CUMS Rats by Protecting Hippocampal Synapse and Mitochondrion: An Ultrastructural and iTRAQ Proteomic Study.

Electroacupuncture (EA) is considered a complementary therapy for depression. Trials also found that EA has additive benefits when combined with medication compared with medication alone. It is revealed that EA restores altered hippocampal synaptic plasticity in depressed brain. But precise molecular mechanism is poorly understood. Here, we evaluated the therapeutic effects of EA and EA combined with selective serotonin reuptake inhibitor (SSRI) on depressed (CUMS) rats. Then a new proteomics approach, isobaric tag for relative and absolute quantitation (iTRAQ), was used to explore the differential expressed synaptic protein in hippocampus between CUMS and EA-treated rats to identify the possible target molecular mechanism of its effects. We found that EA had additive benefit against depressive behaviors when combined with SSRI. Ultrastructure study on neuron showed significant change in postsynapse density (PSD) and mitochondrion. Through iTRAQ, it is found that synaptic and mitochondrial proteins were significantly changed after EA, consisting with ultrastructure study results. These findings suggest that EA improves antidepressant performance in depressed rats through protecting synaptic and mitochondrial functions in hippocampus.

Different Brain Activation after Acupuncture at Combined Acupoints and Single Acupoint in Hypertension Patients: An Rs-fMRI Study Based on ReHo Analysis.

BACKGROUND: Acupuncture is proved to be effective on hypertension by numerous studies and resting-state functional magnetic resonance imaging (Rs-fMRI) is a widely used technique to study its mechanism. Along with lower blood pressure, patients with hypertension receiving acupuncture also presented improvement in function of cognition, emotion, language, sematic sensation, and so on. This study was a primary study to explore the acting path of acupuncture at combined acupoints in stimulated brain areas related to such functions. METHODS: In this research, regional homogeneity (ReHo) was applied to analyze the Rs-fMRI image data of brain activities after acupuncture at LR3, KI3, and LR3+KI3 and to compare the differences of functional brain activities between stimulating combined acupoints and single acupoint under pathological conditions. A total of thirty hypertension patients underwent Rs-fMRI scanning before acupuncture treatment and then were randomly divided into three groups following random number table, the LR3 group (3 males and 7 females), the KI3 group (3 males and 7 females), and the LR3+ KI3 group (4 males and 6 females) for needling, respectively. When the 30-min treatment finished, they received a further Rs-fMRI scanning. The Rs-fMRI data before and after the acupuncture treatment were analyzed through ReHo. RESULTS: Compared with preacupuncture, respectively, ReHo values increased in Brodmann areas (BAs) 3, 18, and 40 and decreased in BAs 7 and 31 in LR3+ KI3 group. However, ReHo values only decreased in BA7 of KI3 group while the results showed no significant difference of brain regions in LR3 group between pre- and postacupuncture. Compared with LR3 group, LR3+KI3 group exhibited decreased ReHo values in BAs 7, 9, and 31. Meanwhile, compared with KI3 group, LR3+KI3 group exhibited increased ReHo values in the BAs 2, 18, 30, and 40 and decreased ReHo values in BA13. CONCLUSION: Combined acupoints of LR3 and KI3 could act on wider brain areas than the sum of single acupoints, whose functions include emotional processing, cognition, somatic sensation, spatial orientation, language production, and vision.

Equilibrium, kinetics and thermodynamics of Cu(II) biosorption on Chinese chestnut shell pretreated with steam explosion.

The shells of Chinese chestnuts (Castanea mollissima) are an agricultural residue. This work aimed to evaluate this feasibility of using steam explosion to modify this residue for Cu(II) biosorption from aqueous solutions. Equilibrium, kinetic and thermodynamic parameters were evaluated. The steam-explosion pretreatment increased the surface area of the chestnut shell and exposed more hydroxyl and carboxyl groups, which are binding sites for Cu(II). It changed the sorption from a spontaneous process driven by enthalpy to a nonspontaneous one driven by entropy. It increased the Cu(II) sorption capacity at higher temperatures while it decreased the capacity at lower ones. Compared with untreated chestnut shell, the steam-exploded shell is preferable for Cu(II) sorption at higher temperatures.

Insolubilization of Chestnut Shell Pigment for Cu(II) Adsorption from Water.

Chestnut shell pigment (CSP) is melanin from an agricultural waste. It has potential as an adsorbent for wastewater treatment but cannot be used in its original state because of its solubility in water. We developed a new method to convert CSP to insolubilized chestnut shell pigment (ICSP) by heating, and the Cu(II) adsorption performance of ICSP was evaluated. The conversion was characterized, and the thermal treatment caused dehydration and loss of carboxyl groups and aliphatic structures in CSP. The kinetic adsorption behavior obeyed the pseudo-second-order rate law, and the equilibrium adsorption data were well described with both the Langmuir and the Freundlich isotherms. ICSP can be used as a renewable, readily-available, easily-producible, environmentally-friendly, inexpensive and effective adsorbent to remove heavy-metal from aquatic environments.

Comparison of Antioxidant Activities of Melanin Fractions from Chestnut Shell.

Chestnut shell melanin can be used as a colorant and antioxidant, and fractionated into three fractions (Fr. 1, Fr. 2, and Fr. 3) with different physicochemical properties. Antioxidant activities of the fractions were comparatively evaluated for the first time. The fractions exhibited different antioxidative potential in different evaluation systems. Fr. 1, which is only soluble in alkaline water, had the strongest peroxidation inhibition and superoxide anion scavenging activity; Fr. 2, which is soluble in alkaline water and hydrophilic organic solvents but insoluble in neutral and acidic water, had the greatest power to chelate ferrous ions; and Fr. 3, which is soluble both in hydrophilic organic solvents and in water at any pH conditions, had the greatest hydroxyl (·OH) and 1,1-diphenyl-2-picryl-hydrazyl (DPPH·) radicals scavenging abilities, reducing power, and phenolic content. The pigment fractions were superior to butylated hydroxytolune (BHT) in ·OH and DPPH· scavenging and to ethylene diamine tetraacetic acid (EDTA) in the Fe(2+)-chelation. They were inferior to BHT in peroxidation inhibition and O2·(-) scavenging and reducing power. However, BHT is a synthetic antioxidant and cannot play the colorant role. The melanin fractions might be used as effective biological antioxidant colorants.

Isolation, fractionation and characterization of melanin-like pigments from chestnut (Castanea mollissima) shells.

UNLABELLED: Melanins are known as versatile biopolymers, but the utilizations are restricted by their poor solubilities. Therefore, well soluble ones or their analogs are much desired. In this article, a new procedure was developed for fractionation of the pigments isolated from chestnut (Castanea mollissima) shells, and 3 fractions (Fr. 1, Fr. 2, and Fr. 3) were obtained. The solubilities of all the fractions in waters of different pH and in common organic solvents were studied. The physicochemical properties of the fractions were characterized for the first time on the basis of combined chemical analyses and spectroscopic methods including ultraviolet-visible (UV-Vis), Fourier transform infrared (FT-IR), electron spin resonance (ESR), and solid-state ¹³C nuclear magnetic resonance (¹³C-NMR). All the fractions could be bleached by NaOCl and H2O2 and give a positive reaction for polyphenols, which are usually used as typical tests for allomelanins. Their UV-Vis, FT-IR, and ESR spectra resembled those of synthetic and some natural melanins. Elemental data and quantitative analyses of ¹³C-NMR spectra revealed that pigment-bound proteins and polysaccharides were the most abundant in Fr. 1, while Fr. 2 was presented with the highest aromaticity. PRACTICAL APPLICATION: We provided a new, simple, and inexpensive method to fractionate the melanin-like pigments from chestnut shells. This technique can be used to produce natural melanin-like food colorants with different solubilities from chestnut shells.

Histological observation of somatic embryogenesis from cultured embryos of Quercus variabilis Bl.

Immature zygotic embryos of Quercus variabilis Bl. were excised and cultured on MS basal medium containing 0.25 mg/L 2,4-dichlorophenoxyacetic acid and 0.5 mg/L 6-benzyl aminopurine. Callus was initiated from these embryos within 6 weeks. Two types of embryogenic calluses were formed: one was the white mucilaginous callus and the other was light yellow to translucent, glossy, mucilaginous. Histological examination showed that the non-embryogenic cells derived from zygotic embryos were large in size, with small nuclei, thin cytoplasm, and the embryogenic cells were small in size, with large nuclei, thick cytoplasm and denser arrangement. Somatic embryos were induced from embryogenic masses (EMS) cultured on a medium not containing plant growth regulators (PGRs). The histological origin of somatic embryos was single epidermal or subepidermal cells. Somatic embryos displayed a standard development pattern, from globular ones to heart-like and torpedo-shaped ones and finally to embryo with two cotyledons, which was similar to the development of zygotic embryo in vivo. Secondary proembryos were formed on the axis and cotyledon of existing embryos and originated from single, densely stained cells of the epidermis at all stages.