[Effect of Tongdu Tiaoshen acupuncture on CREB/BDNF/TrkB signaling pathway of hippocampus in rats with post-stroke depression].

OBJECTIVE: To observe the regulative effect of Tongdu Tiaoshen acupuncture on the depression-like behavior and cAMP-response element binding protein (CREB)/brain-derived neurotrophic factor (BDNF)/tyrosine protein kinase B (TrkB) signaling pathway of hippocampus in rats with post-stroke depression (PSD), and to explore its possible mechanism on improving PSD. METHODS: A total of 36 SPF SD rats were randomized into a sham operation group, a model group and a Tongdu Tiaoshen group, 12 rats in each group. The compound method of Zea Longa suture-occlusion and chronic unpredictable mild stress (CUMS) was used to establish the PSD model in rats of the model group and the Tongdu Tiaoshen group. On the 4th day after modeling, acupuncture was applied at "Dazhui" (GV 14), "Shuigou" (GV 26), "Baihui" (GV 20) and "Shenting" (GV 24) in the Tongdu Tiaoshen group, 40 min every time, once a day, 6 times a week for 4 weeks consecutively. On the 2nd day after PSD modeling and after 4-week intervention, Zea Longa neurobehavioral score was evaluated, sucrose water consumption test and open-field test were performed; biochemical method was used to detect the SOD, CAT activity and MDA level in hippocampal CA1 area; ELISA method was used to detect the serum level of BDNF; real-time PCR was used to detect the mRNA expression of BDNF, TrkB and CREB in hippocampal CA1 area; Western blot was used to detect the protein expression of BDNF, TrkB, CREB and p-CREB in hippocampal CA1 area. RESULTS: Compared with the sham operation group, Zea Longa neurobehavioral scores were increased (P<0.05), percentage of sucrose water consumption, horizontal motion and vertical motion scores of open-field test were decreased after modeling and intervention in the model group and after modeling in the Tongdu Tiaoshen group (P<0.05). Compared with the model group, Zea Longa neurobehavioral score was decreased (P<0.05), percentage of sucrose water consumption, horizontal motion and vertical motion scores of open-field test were increased after intervention in the Tongdu Tiaoshen group (P<0.05). Compared with the sham operation group, the SOD and CAT activity in hippocampal CA1 area and serum level of BDNF were decreased (P<0.05), MDA level in hippocampal CA1 area was increased in the model group (P<0.05); compared with the model group, the SOD and CAT activity in hippocampal CA1 area and serum level of BDNF were increased (P<0.05), MDA level was decreased in the Tongdu Tiaoshen group (P<0.05). Compared with the sham operation group, the mRNA expression of BDNF, TrkB and CREB as well as the protein expression of BDNF, TrkB, CREB and p-CREB were decreased in hippocampal CA1 area in the model group (P<0.05); compared with the model group, the mRNA expression of BDNF, TrkB and CREB, the protein expression of BDNF, TrkB and p-CREB as well as the ratio of p-CREB/CREB were increased in the Tongdu Tiaoshen group (P<0.05). CONCLUSION: Tongdu Tiaoshen acupuncture can improve the depression-like behavior in PSD rats, the mechanism may be related to the inhibition of oxidative stress in hippocampal tissues and the enhanced activity of CREB/BDNF/TrkB signaling pathway.

[Effect of Tongdu Tiaoshen acupuncture on PI3K/Akt/mTOR signaling pathway and autophagy-related proteins of hippocampus in rats with post-stroke depression].

OBJECTIVE: To observe the improvement of Tongdu Tiaoshen acupuncture on depression-like behavior in rats with post-stroke depression (PSD), and to explore its possible mechanism. METHODS: A total of 48 SD rats were randomized into a sham-operation group, a model group, a Tongdu Tiaoshen group and a non-meridian-non-acupoint group, 12 rats in each group. In the the model group, the Tongdu Tiaoshen group and the non-meridian-non-acupoint group, PSD model was established by the compound method of cerebral medial arterial occlusion, chronic unpredictable mild stress and isolation. Acupuncture was applied at "Dazhui"(GV 14), "Shuigou"(GV 26), "Baihui"(GV 20) and "Shenting" (GV 24) in the Tongdu Tiaoshen group, and at the bilateral dorsum hollows between 3rd and 4th metatarsal bones of the forelegs in the non-meridian-non-acupoint group, needles were retained for 40 min in the both groups, once a day, 6 times a week for 4 weeks. Before model establishment and after intervention, the ethology indice of Zea Longa neurological behavioral score, open-field test and sucrose preference test were evaluated in the 4 groups. The levels of phosphatidylinositol 3-kinase (PI3K), phosphorylated protein kinase B (p-Akt), phosphorylated mammalian target of rapamycin (p-mTOR), Beclin1 and LC3B-Ⅱ/Ⅰin the CA1 region of hippocampus were detected by western blot. The expressions of Beclin1, LC3B-ⅡmRNA in the CA1 region of hippocampus were detected by real-time PCR. RESULTS: After model established, compared with the sham-operation group, the Zea Longa neurological behavioral scores were increased (P<0.05), the scores of horizontal and vertical motion of the open-field test and the consumption of sucrose solution were decreased in the model group, the Tongdu Tiaoshen group and the non-meridian-non-acupoint group (P<0.05). After intervention, compared with the sham-operation group, the Zea Longa neurological behavioral score, the expression of Beclin1, LC3B-Ⅱ/Ⅰ and LC3B-Ⅱin the CA1 region of hippocampus were increased (P<0.05), the scores of horizontal and vertical motion of the open-field test, the consumption of sucrose solution and the expression of PI3K, p-Akt and p-mTOR were decreased in the model group (P<0.05). After intervention, compared with the model group and the non-meridian-non-acupoint group, the Zea Longa neurological behavioral score, the expression of Beclin1, LC3B-Ⅱ/Ⅰ and LC3B-Ⅱin the CA1 region of hippocampus were decreased (P<0.05), the scores of horizontal and vertical motion of the open-field test, the consumption of sucrose solution and the expression of PI3K, p-Akt and p-mTOR were increased in the Tongdu Tiaoshen group (P<0.05). CONCLUSION: Tongdu Tiaoshen acupuncture can effectively improve the depression-like behavior in PSD rats, its mechanism may relate to the activation of PI3K/Akt/mTOR signaling pathway and the inhibition of hippocampal neuron autophagy.

[Professor CHU Hao-ran's experience of acupuncture and moxibustion of regulating spleen-stomach and differentiating meridians-collaterals].

Professor CHU Hao-ran successively followed famous doctors of acupuncture and spleen-stomach disease, such as ZHOU Mei-sheng, inherited their academic thoughts and clinical experience, and studied the classics to summarize and propose the clinical academic proposition regulating spleen-stomach, differentiating meridians-collaterals. In clinical practice, professor CHU focuses on individual differences of patients, carefully examines syndrome and refines acupoint selection; he pays attention to regulating spleen-stomach, and actively uses Zusanli (ST 36); he simultaneously uses acupuncture-moxibustion and acupoints with efficacy; he inherits traditional reinforcing and reducing methods, and extends the scope of acupoint diagnosis and treatment, while he emphasizes keeping spirit and treating spirit, and the combination of mind and qi, in order to improve clinical diagnosis and treatment effect.

[Protective effects on hippocampal neurons and the influence on hippocampal monoamine neurotransmitters with acupuncture for promoting the circulation of the governor vessel and regulating the mental state in rats with post-stroke depression].

OBJECTIVE: To observe the effects on hippocampal neuronal structure and hippocampal monoamine neurotransmitters such as norepinephrine (NE), 5-hydroxytryptamine (5-HT) and dopamine (DA) with acupuncture for promoting the circulation of the governor vessel and regulating the mental state (Tongdu Tiaoshen acupuncture) in rats with post-stroke depression and explore the underlying effect mechanism of Tongdu Tiaoshen acupuncture on post-stroke depression. METHODS: A total of 48 clean-grade SD rats were randomized into a sham-operation group, a model group, a Tongdu Tiaoshen acupuncture group and a non-meridian-non-acupoint group, 12 rats in each one. In the model group, the Tongdu Tiaoshen acupuncture group and the non-meridian-non-acupoint group, the compound method of cerebral medial arterial occlusion and the chronic unpredictable mild stress was adopted to prepare the model of post-stroke depression. In the sham-operation group, skin was opened and the fascia muscle was separated, after artery exposed, no ligation was operated and the incision was sutured directly. In the sham-operation group and the model group, no any intervention was performed. In the Tongdu Tiaoshen acupuncture group, acupuncture was applied to "Baihui" (GV 20), "Shuigou" (GV 26), "Shenting" (CV 24) and "Dazhui" (GV 14) for 40 min in each treatment and the needles were manipulated at the acupoints once in 20 min. The treatment was given once a day, 6 times a week for 4 weeks consecutively. In the non-meridian-non-acupoint group, the depression between the 3rd and 4th metatarsal bones on the dorsum of the forelimb was selected in intervention and the needling technique was same as the Tongdu Tiaoshen acupuncture group. The behavioral changes were observed after intervention in each group. Using transmission electron microscopy, the ultrastructure of hippocampal CA 1 neurons was observed in the rats of each group. Using high performance liquid chromatography, the contents of NE, 5-HT and DA were determined. RESULTS: Compared with the sham-operation group, the Zea-longa neurobehavioral score was increased (P<0.01), the sucrose water consumption was reduced (P<0.01), the number of times of horizontal movement and vertical movement in open-field test were reduced (both P<0.01), the damage of the ultrastructure of hippocampal CA 1 neurons was aggravated and the contents of NE, 5-HT and DA were reduced in the model group (P<0.01). Compared with the model group, the Zea-Longa neurobehavioral score was reduced (P<0.01), the sucrose water consumption was increased (P<0.01), the number of times of horizontal movement and vertical movement in open-field test were increased (both P<0.01), the damage of the ultrastructure of hippocampal CA 1 neurons was alleviated and the contents of NE, 5-HT and DA were increased in the Tongdu Tiaoshen acupuncture group (P<0.01, P<0.05). Compared with the non-meridian-non-acupoint group, the Zea-Longa neurobehavioral score was reduced (P<0.01), the sucrose water consumption was increased (P<0.01), the number of times of horizontal movement and vertical movement in open-field test were increased (both P<0.01), the damage of the ultrastructure of hippocampal CA 1 neurons was alleviated and the contents of NE, 5-HT and DA were increased in the Tongdu Tiaoshen acupuncture group (P<0.01, P<0.05). CONCLUSION: The Tongdu Tiaoshen acupuncture improves the behavior of the model rats with post-stroke depression, repairs hippocampal neuronal damage, which is probably related to the contents of hippocampal monoamine neurotransmitters (NE, 5-HT and DA).

Retracted Article: A high-throughput metabolomics strategy for discovering the influence of differential metabolites and metabolic pathways of huaxian capsules on sepsis-associated Qi deficiency and blood stasis syndrome.

High-throughput metabolomics can be used to investigate the therapeutic targets and metabolic mechanisms of traditional Chinese medicine (TCM) formulae, which have multiple targets in disease therapy, but it is a great challenge to explore their mechanism of action. Huaxian capsule (HXC) is a classical formula in TCM that has therapeutic effects on a sepsis-associated Qi deficiency and blood stasis syndrome (SQBS). However, its targets and metabolic mechanisms need more investigation. To investigate the therapeutic effects of HXC in the treatment of SQBS and elucidate the potential mechanism, we used a high-throughput metabolomics strategy based on the ultraperformance liquid chromatography/mass spectrometry combined with chemometrics to analyze and identify differential metabolites and pathways. The pathological examination of organs and biochemical indices was also performed to verify the successful establishment of the rat model and protective effects of HXC. Pathological symptoms and biochemical indicators of SQBS rats were reversed by the HXC treatment. A total of 24 potential biomarkers were identified to indicate the difference between the control and model groups; they were closely associated with ten metabolic pathways and regulated by the HXC administration. From the pathway analysis, we further understood the protective activity of HXC against SQBS, which affected amino acid metabolism, molecular transport, small molecule biochemistry and cell signaling as well as vitamin and mineral metabolism. In conclusion, HXC protects against SQBS by modulating the metabolic biomarkers and functional pathways.