Artesunate improves glucose and lipid metabolism in db/db mice by regulating the metabolic profile and the MAPK/PI3K/Akt signalling pathway.

BACKGROUND: Diabetes is a metabolic disorder characterized by chronic hyperglycaemia. Chronic metabolic abnormalities and long-term hyperglycaemia may result in a wide range of acute and chronic consequences. Previous studies have demonstrated that artesunate(ART) has antidiabetic, anti-inflammatory, antiatherosclerotic, and other beneficial effects, but the specific regulatory mechanism is not completely clear. AIM: This study investigated the effects of ART on metabolic disorders in type 2 diabetes mellitus (T2DM) model db/db mice and explored the underlying mechanisms involved. METHODS: C57BL/KsJ-db/db mice were used to identify the targets and molecular mechanism of ART. Metabolomic methods were used to evaluate the efficacy of ART in improving T2DM-related metabolic disorders. Network pharmacology and transcriptomic sequencing were used to analyse the targets and pathways of ART in T2DM. Finally, molecular biology experiments were performed to verify the key targets and pathways selected by network pharmacology and transcriptomic analyses. RESULTS: After a 7-week ART intervention (160 mg/kg), the glucose and lipid metabolism levels of the db/db mice improved. Additionally, the oxidative stress indices, namely, the MDA and SOD levels, significantly improved (p<0.01). Linoleic acid and glycerophospholipid metabolism, amino acid metabolism, bile acid synthesis, and purine metabolism disorders in db/db mice were partially corrected after ART treatment. Network pharmacology analysis identified important targets of ART for the treatment of metabolic disorders in T2DM . These targets are involved in key signalling pathways, including the highest scores observed for the PI3K/Akt signalling pathway. Transcriptomic analysis revealed that ART could activate the MAPK signalling pathway and two key gene targets, HGK and GADD45. Immunoblotting revealed that ART increases p-PI3K, p-AKT, Glut2, and IRS1 protein expression and suppresses the phosphorylation of p38, ERK1/2, and JNK, returning HGK and GADD45 to their preartesunate levels. CONCLUSION: Treatment of db/db mice with 160 mg/kg ART for 7 weeks significantly reduced fasting blood glucose and lipid levels. It also improved metabolic imbalances in amino acids, lipids, purines, and bile acids, thereby improving metabolic disorders. These effects are achieved by activating the PI3K/AKT pathway and inhibiting the MAPK pathway, thus demonstrating the efficacy of the drug.

FMRI connectivity analysis of acupuncture effects on the whole brain network in mild cognitive impairment patients.

The increased risk for the elderly with mild cognitive impairment (MCI) to progress to Alzheimer's disease makes it an appropriate condition for investigation. While the use of acupuncture as a complementary therapeutic method for treating MCI is popular in certain parts of the world, the underlying mechanism is still elusive. We sought to investigate the acupuncture effects on the functional connectivity throughout the entire brain in MCI patients compared to healthy controls (HC). The functional magnetic resonance imaging experiment was performed with two different paradigms, namely, deep acupuncture (DA) and superficial acupuncture (SA), at acupoint KI3. We first identified regions showing abnormal functional connectivity in the MCI group compared to HC during the resting state and subsequently tested whether these regions could be modulated by acupuncture. Then, we made the comparison of MCI vs. HC to test whether there were any specific modulatory patterns in the poststimulus resting brain between the two groups. Finally, we made the comparisons of DA vs. SA in each group to test the effect of acupuncture with different needling depths. We found the temporal regions (hippocampus, thalamus, fusiform gyrus) showing abnormal functional connectivity during the resting state. These regions are implicated in memory encoding and retrieving. Furthermore, we found significant changes in functional connectivity related with the abnormal regions in MCI patients following acupuncture. Compared to HC, the correlations related with the temporal regions were enhanced in the poststimulus resting brain in MCI patients. Compared to SA, significantly increased correlations related with the temporal regions were found for the DA condition. The enhanced correlations in the memory-related brain regions following acupuncture may be related to the purported therapeutically beneficial effects of acupuncture for the treatment of MCI. The heterogeneous modulatory patterns between DA and SA may suggest that deep muscle insertion of acupuncture is necessary to achieve the appreciable clinical effect.

Investigation of acupoint specificity by multivariate granger causality analysis from functional MRI data.

PURPOSE: To investigate the acupoint specificity by exploring the effective connectivity patterns of the poststimulus resting brain networks modulated by acupuncture at the PC6, with the same meridian acupoint PC7 and different meridian acupoint GB37. MATERIALS AND METHODS: The functional MRI (fMRI) study was performed in 36 healthy right-handed subjects receiving acupuncture at three acupoints, respectively. Due to the sustained effects of acupuncture, a novel experimental paradigm using the nonrepeated event-related (NRER) design was adopted. Psychophysical responses (deqi sensations) were also assessed. Finally, a newly multivariate Granger causality analysis (mGCA) was used to analyze effective connectivity patterns of the resting fMRI data taken following acupuncture at three acupoints. RESULTS: Following acupuncture at PC6, the red nucleus and substantia nigra emerged as central hubs, in comparison with the fusiform gyrus following acupuncture at GB37. Red nucleus was also a target following acupuncture at PC7, but with fewer inputs than those of PC6. In addition, the most important target following acupuncture at PC7 was located at the parahippocampus. CONCLUSION: Our findings demonstrated that acupuncture at different acupoints may exert heterogeneous modulatory effects on the causal interactions of brain areas during the poststimulus resting state. These preliminary findings provided a clue to elucidate the relatively function-oriented specificity of acupuncture effects.

Neural specificity of acupuncture stimulation from support vector machine classification analysis.

Acupoint specificity, as a crucial issue in acupuncture neuroimaging studies, is still a controversial topic. Previous studies have generally adopted a block-based general linear model (GLM) approach, which predicts the temporal changes in the blood oxygenation level-dependent signal conforming to the "on-off" specifications. However, this method might become impractical since the precise timing and duration of acupuncture actions cannot be specified a priori. In the current study, we applied a data-driven multivariate classification approach, namely, support vector machine (SVM), to explore the neural specificity of acupuncture at gall bladder 40 (GB40) using kidney 3 (KI3) as a control condition (belonging to different meridians but the same nerve segment). In addition, to verify whether the typical GLM approach is sensitive enough in exploring the neural response patterns evoked by acupuncture, we also employed the GLM method to the same data sets. The SVM analysis detected distinct neural response patterns between GB40 and KI3--positive predominantly for the GB40, while negative following the KI3. By contrast, group analysis from the GLM showed that acupuncture at these different acupoints can both evoke similar widespread signal decreases in multiple brain regions, and most of these regions were spatially overlapped, mainly distributing in the limbic and subcortical structures. Our findings may provide additional evidence to support the specificity of acupuncture, relevant to its clinical efficacy. Moreover, we also proved that GLM analysis is prone to be susceptible to errors and is not appropriate for detecting neural response patterns evoked by acupuncture stimulation.

Investigation of the large-scale functional brain networks modulated by acupuncture.

Previous neuroimaging studies have primarily focused on the neural activities involving the acute effects of acupuncture. Considering that acupuncture can induce long-lasting effects, several researchers have begun to pay attention to the sustained effects of acupuncture on the resting brain. Most of these researchers adopted functional connectivity analysis based on one or a few preselected brain regions and demonstrated various function-guided brain networks underlying the specific effect of acupuncture. Few have investigated how these brain networks interacted at the whole-brain level. In this study, we sought to investigate the functional correlations throughout the entire brain following acupuncture at acupoint ST36 (ACUP) in comparison with acupuncture at nearby nonacupoint (SHAM). We divided the whole brain into 90 regions and constructed functional brain network for each condition. Then we examined the network hubs and identified statistically significant differences in functional correlations between the two conditions. Following ACUP, but not SHAM, the limbic/paralimbic regions such as the amygdala, hippocampus and anterior cingulate gyrus emerged as network hubs. For direct comparisons, increased correlations for ACUP compared to SHAM were primarily related with the limbic/paralimbic and subcortical regions such as the insula, amygdala, anterior cingulate gyrus, and thalamus, whereas decreased correlations were mainly related with the sensory and frontal cortex. The heterogeneous modulation patterns between the two conditions may relate to the functional specific modulatory effects of acupuncture. The preliminary findings may help us to better understand the long-lasting effects of acupuncture on the entire resting brain, as well as the neurophysiological mechanisms underlying acupuncture.

Investigation of acupoint specificity by whole brain functional connectivity analysis from fMRI data.

Previous neuroimaging studies on acupuncture have primarily adopted functional connectivity analysis associated with one or a few preselected brain regions. Few have investigated how these brain regions interacted at the whole brain level. In this study, we sought to investigate the acupoint specificity by exploring the whole brain functional connectivity analysis on the post-stimulus resting brain modulated by acupuncture at acupoint PC6, with the same meridian acupoint PC7 and different meridian acupoint GB37. We divided the whole brain into 90 regions and analyzed functional connectivity for each condition. Then we identified statistically significant differences in functional correlations throughout the entire brain following acupuncture at PC6 in comparison with PC7 as well as GB37. For direct comparisons, increased correlations for PC6 compared to PC7 were primarily between the prefrontal regions and the limbic/paralimbic and subcortical regions, whereas decreased correlations were mainly between the parietal regions and the limbic/paralimbic and subcortical regions. On the other hand, increased correlations for PC6 compared to GB37 were primarily between the prefrontal regions and somatosensory regions, whereas decreased correlations were mainly related with the occipital regions. Our findings demonstrated that acupuncture at different acupoints may exert heterogeneous modulatory effects on the post-stimulus resting brain, providing new evidences for the relatively function-oriented specificity of acupuncture effects.

Investigation of acupoint specificity by functional connectivity analysis based on graph theory.

Acupoint specificity is still a contentious issue and remains to be verified whether brain, response is as specific as the purported indication of different acupoints. Previous fMRI acupuncture, studies based on multiple-block design may not be able to fully disclose acupuncture effects. Both, recent studies and certain clinical reports have indicated that there exists time-sustainability during, acupuncture even after the stimulus being terminated. Further understanding of how such external, intervention interacts with post-stimulus resting brain networks may enlighten us to gain an, appreciation of the physiological function and integrated mechanisms involved in acupuncture. In our, study, we adopted a modified non-repeated event-related (NRER) design, and utilized the graph theory, based functional connectivity analysis to investigate the neural specificity of the PC6, with the same, meridian acupoint PC7 and a different meridian acupoint GB37 as separate controls. Under the, construct of this network model, some brain regions with a larger degree of connectivity indicated, stronger interactions with other brain regions and were considered to be important nodes in this, network. We identified that the two most important brain areas were the right nodule and right uvula, following acupuncture at PC6, and the right amygdala and right inferior parietal lobe following, acupuncture at PC7. Following the GB37, the two regions with the larger degree of connectivity were, the posterior cingulate cortex (PCC) and middle occipital gyrus. These specific regions may mediate the, specific effects of acupuncture. Results showed that different modulatory brain networks may support, the point specificity of acupuncture.

The salient characteristics of the central effects of acupuncture needling: limbic-paralimbic-neocortical network modulation.

Human and animal studies suggest that acupuncture produces many beneficial effects through the central nervous system. However, the neural substrates of acupuncture actions are not completely clear to date. fMRI studies at Hegu (LI4) and Zusanli (ST36) indicated that the limbic system may play an important role for acupuncture effects. To test if this finding applies to other major classical acupoints, fMRI was performed on 10 healthy adults during manual acupuncture at Taichong (LV3), Xingjian (LV2), Neiting (ST44), and a sham point on the dorsum of the left foot. Although certain differences could be observed between real and sham points, the hemodynamic response (BOLD signal changes) and psychophysical response (sensory experience) to acupuncture were generally similar for all four points. Acupuncture produced extensive deactivation of the limbic-paralimbic-neocortical system. Clusters of deactivated regions were seen in the medial prefrontal cortex (frontal pole, pregenual cingulate), the temporal lobe (amygdala, hippocampus, and parahippocampus) and the posterior medial cortex (precuneus, posterior cingulate). The sensorimotor cortices (somatosensory cortices, supplementary motor cortex), thalamus and occasional paralimbic structures such as the insula and anterior middle cingulate cortex showed activation. Our results provide additional evidence in support of previous reports that acupuncture modulates the limbic-paralimbic-neocortical network. We hypothesize that acupuncture may mediate its antipain, antianxiety, and other therapeutic effects via this intrinsic neural circuit that plays a central role in the affective and cognitive dimensions of pain as well as in the regulation and integration of emotion, memory processing, autonomic, endocrine, immunological, and sensorimotor functions.