NGF/PI3K/TRPV1 pathway mediates the regulation of visceral pain by acupuncture at "Shangjuxu" (ST37) in irritable bowel syndrome rats with chronic visceral hyperalgesia. / NGF/PI3K/TRPV1通路介导针刺"上巨虚"è°ƒèŠ‚è‚ æ˜“æ¿€ç»¼åˆå¾æ ¢æ€§å† è„ç—›æ•æ¨¡åž‹å¤§é¼ å† è„ç—›.

OBJECTIVES: To investigate the effect of manual acupuncture at "Shangjuxu"(ST37) on nerve growth factor(NGF)/phosphatidylinositol 3-kinase(PI3K)/transient receptor potential vanilloid 1(TRPV1) signaling pathway in rats with chronic visceral hyperalgesia of irritable bowel syndrome (IBS), so as to explore its underlying mechanism in treating IBS chronic visceral hyperalgesia. METHODS: IBS chronic visceral hyperalgesia model was established by colorectal dilation stimulation for 2 weeks for SD pups at 8 d after birth, which were fed until 8-week age after the stimulation. Then the verified successfully modeled adult rats were randomly divided into model, Shangjuxu, and non-acupoint groups, with 6 rats in each group, and 6 unmodeled rats were selected as normal group. On the next day of model evaluation, rats in the Shangjuxu group received acupuncture at right ST37 while rats in the non-acupoint group received acupuncture at the non-meridian and non-acupoint point in the right hypochondrium, both for 15 min, with manual twisting of mild reinforcing and reducing performed for 30 s at an interval of 5 min, once a day, for a total of 7 d. Abdominal withdrawal reflex(AWR) score was used to evaluate the degree of chronic visceral pain in rats. Western blot and real-time fluorescence quantitative PCR were used to detect the colonic protein and mRNA expressions of NGF, tropomyosin receptor kinase A (TrkA), PI3K and TRPV1. The positive expressions of PI3K and TRPV1 proteins in the colon of rats were detected by immunohistochemistry method. RESULTS: Compared with the normal group, AWR scores corresponding to 4 pressure levels of 20, 40, 60 and 80 mm Hg, mRNA and protein expressions of NGF, TrkA, PI3K and TRPV1 in colon tissue, and positive expressions of PI3K and TRPV1 in colon tissue were significantly increased(P<0.05) in the model group. After intervention, compared with the model group, rats in the Shangjuxu group had reduced AWR scores corresponding to 4 pressure levels of 20, 40, 60 and 80 mm Hg, lower colonic mRNA and protein expressions of NGF, TrkA, PI3K and TRPV1, and decreased positive expressions of PI3K and TRPV1 in colon tissue(P<0.05), while there were no significant differences in the above indexes of the non-acupoint group. CONCLUSIONS: Manual acupuncture at ST37 can alleviate IBS chronic visceral hyperalgesia in rat and its analgesic effect may be related to regulating NGF/PI3K/TRPV1 signaling pathway.

[Effect of transcutaneous electrical acupoint stimulation on learning and memory ability of chronic fatigue syndrome rats and its mechanisms].

OBJECTIVE: To observe the effect of transcutaneous electrical acupoint stimulation (TEAS) on the histomorphological manifestations of hippocampal CA1 region and the expression of extracellular regulatory protein kinase (ERK), cyclic adenosine response element binding protein (CREB) and brain-derived neurotrophic factor (BDNF) in chronic fatigue syndrome (CFS) rats, so as to explore the mechanisms of TEAS in improving the learning and memory abilities of CFS rats. METHODS: Forty male Wistar rats were randomly divided into normal group (10 rats) and modeling group (30 rats); then after modeling, they were selected and randomly divided into model group (10 rats) and TEAS group (10 rats). CFS rats model was prepared by sleep deprivation combined with weight-bearing swimming. Rats in the TEAS group were stimulated with Han's acupoint nerve stimulator at bilateral "Zusanli" (ST36) and "Shenshu" (BL23) (2 Hz/15 Hz, 1-2 mA), 20 min each time, once a day for 4 weeks with 1 d rest every 6 d. The score of general conditions of rats was evaluated. The learning and memory ability was tested with Morris water maze. The morphology and ultrastructure of hippocampal CA1 region were observed by HE staining and transmission electron microscopy. The expression levels of ERK, CREB and BDNF mRNAs and proteins in hippocampus were detected by real time quantitative PCR and Western blot, respectively. RESULTS: Compared with the normal group, the score of general condition was increased (P<0.01); the escape latency was prolonged (P<0.05, P<0.01) and the times of crossing the original platform was decreased (P<0.05); the expression levels of ERK, CREB and BDNF mRNAs and proteins in hippocampus were decreased (P<0.05, P<0.01) in the model group. Compared with the model group, the scores of general condition on the 42nd and 49th day were decreased (P<0.05, P<0.01); the escape latency was shortened (P<0.01, P<0.05)and the times of crossing the original platform were increased (P<0.05); the expression levels of ERK, CREB and BDNF mRNAs and proteins in hippocampus were increased (P<0.01, P<0.05) in the TEAS group. The morphology of neurons in hippocampal CA1 region was normal in the normal group. In the model group, the number of neurons in hippocampal CA1 region decreased, the arrangement of nerve cells was scattered, the number of apoptotic cells increased, some nuclear structures disappeared, nuclear heterochromatin increased, the cell membrane wrinkled, the chromatin appeared empty bright area, and the crista was incomplete. Compared with the model group, the nerve cells morphology in hippocampal CA1 region was more regular, the number of apoptotic cells decreased, the chromatin and the cytoplasm were uniformly distributed, and the crista was relatively intact in the TEAS group. CONCLUSION: TEAS can improve the learning and memory ability of CFS rats, the mechanisms may be related to improving the neural structure of hippocampal CA1 region and up-regulating the expression levels of ERK/CREB/BDNF.

Manual Acupuncture at ST37 Modulates TRPV1 in Rats with Acute Visceral Hyperalgesia via Phosphatidylinositol 3-Kinase/Akt Pathway.

Acupuncture can significantly ameliorate inflammatory pain in acute visceral hyperalgesia. Hyperalgesia is attenuated by inflammatory mediators that activate transient receptor potential vanilloid 1 (TRPV1), and TRPV1 is regulated by nerve growth factor (NGF)-induced phosphatidylinositol 3-kinase (PI3K)/Akt pathway. However, it is unknown whether NGF-induced PI3K/Akt pathway is associated with manual acupuncture (MA). In this study, the effect and mechanism of MA at Shangjuxu (ST37) and Quchi (LI11) were examined using an acetic acid-induced rat model with visceral hyperalgesia. We demonstrated that MA at ST37 significantly decreased abdominal withdrawal reflex (AWR) scores, proinflammatory cytokine expression (TNF-α, IL-1ß, and IL-6), and TRPV1 protein and mRNA expression in rats with acute visceral hyperalgesia compared with the untreated controls, while MA at LI11 showed no effect. The effects of MA at ST37 were reversed after treatment with the PI3K agonist IGF-1 30 min before MA. In rats with visceral hyperalgesia, the upregulation of NGF, tropomyosin-receptor-kinase A (TrkA), PI3K, and phosphorylation-Akt (p-Akt) was decreased by MA at ST37, indicating that TRPV1 regulation via the NGF-induced PI3K/Akt pathway plays a vital role in the effects of MA-mediated amelioration of acute visceral hyperalgesia.

[Analysis on the distribution of pressure- and heat-sensitive acupoints in patients with allergic rhinitis].

OBJECTIVE: To explore the distribution of pressure- and heat-sensitive acupoints of the Large Intestine of Hand-Yangming, Stomach Meridian of Foot-Yangming and the Lung Meridian of Hand-Taiyin below the elbow and knee joints in patients with allergic rhinitis, so as to explore the positive reaction points at the body surface of allergic rhinitis. METHODS: Thirty variant rhinitis volunteer subjects and 30 healthy subjects were recruited in the present study. The subcutaneous induration, reaction points of emptiness-like feeling or pressing pain below the elbow- and knee-joints along the Large Intestine Meridian, Stomach Meridian and the Lung Meridian were inspected first, then re-examined by using heat-sensitized moxibustion method and a hand-held pain threshold tester (mechanical pressure), respectively. The frequency and coincidence rate of pressing-pain sensitive and heat-sensitive points were analyzed. RESULTS: In the 30 patients with allergic rhinitis, 10 positive reaction points as Kongzui (LU6), Chize (LU5), Hegu (LI4), Quchi (LI11), Shousanli(LI10), Zusanli (ST36), Shangjuxu (ST37), Fenglong (ST40), Xiajuxu (ST39), and non-acupoint of the Lung Meridian were found. Among them, the duration of heat-sensitive reaction in the LU6, LI10, ST37, ST40, and ST39 regions was obviously shorter in patients than in healthy subjects (P<0.01,P<0.005). The heat-sensitive forms of the positive reaction points were mainly heat transmission, heat-penetration and local reaction. The mechanical pain thresholds of the positive reaction points (LI4, LU6, LI10, LI11 and non-acupoint of the Lung Meridian) at the upper limbs and those (ST36, ST37 and ST40) at the lower limbs were evidently lower in patients than in healthy volunteer subjects (P<0.05). The pressure- and heat-sensitive acupoints were basically coincide in the patients, but being higher in patients than in healthy subjects in the frequency of occurrence. The mechanical pain threshold of the pressure-sensitive acupoints in patients was remarkably lower on the left side than on the right side of the upper limb, and obviously higher on the left side than on the right side of the lower limb (P<0.05). No significant differences were found between the right and left sides in the pain threshold in healthy subjects (P>0.05). CONCLUSION: There is a high overlap in the distribution of pressure- and heat-sensitive acupoints of the Lung, Large Intestine and Stomach meridians below the elbow and knee joints in patients with allergic rhinitis, and there is an imbalance between the left and right limbs in the mechanical pain threshold of the pressure- and heat-sensitive acupoints, but needing being further confirmed.

Effectiveness and safety of warm needle acupuncture on insomnia in climacteric women: Protocol for a systematic review and meta-analysis.

BACKGROUND: Warm needle acupuncture (WNA) is a traditional Chinese medicine (TCM) therapy which combines technical advantages of acupuncture and moxibustion. Climacteric insomnia is a common symptom in climacteric women, which can seriously affect the physical and mental health of patients. Relevant studies have been reported that WNA can improve insomnia in climacteric women. In this protocol, the effectiveness and safety of WNA on insomnia in climacteric women will be explored. METHODS: Seven electronic databases include 3 English databases [Excerpta Medica database (EMBASE), PubMed, the Cochrane Central Register of Controlled Trials (Cochrane Library)] and 4 Chinese databases [Chinese VIP Information, Chinese National Knowledge Infrastructure (CNKI), Chinese Biomedical Literature Database (CBM) and Wanfang Database] for randomised controlled trials (RCT) of WNA on insomnia in climacteric women will be searched. The changes of the Pittsburgh sleep quality index was used as the main outcome, and the secondary outcome includes the changes of the Kupperman score, serum hormone level, and TCM syndrome score, as well as the adverse events caused by WNA. We will use RevMan software V5.3 to help us to analyze all data and use a Cochrane risk of bias tool to help us to assess the methodological quality for RCTs. RESULT: This study will provide reliable evidence for WNA on insomnia in climacteric women CONCLUSION:: The findings will be an available reference to evaluate the effectiveness and safety of WNA on insomnia in climacteric women. REGISTRATION: PROS-PERO CRD42019125743.

Long-term effects of high lipid and high energy diet on serum lipid, brain fatty acid composition, and memory and learning ability in mice.

OBJECTIVE: It is well known that high lipid and high energy diet is harmful to health. But the different effects of high lipid diet composed of either saturated fatty acids or unsaturated fatty acids have not been distinguished. METHOD: Eighteen pregnant C57BL/6j (22-25g) mice were randomly divided into three groups of six each and fed with chow or high lipid diet composed of either flaxseed oil (chow diet 84%, cholesterol 0.2%, flaxseed oil 15.8%) or lard fat (chow diet 84%, cholesterol 0.2%, lard fat 15.8%). After weaning, the offspring were fed the same diet as their mothers were fed during the experiment, and their spatial memory and learning ability were evaluated by Morris water maze when they were 8 weeks old. Next, the blood and tissues were sampled when they were 9 weeks old. Serum lipids were determined using kits, and brain fatty acids were measured using a gas chromatograph. RESULTS: Compared to chow diet (control), high flaxseed oil diet (HFO) increased high density lipoprotein cholesterol level (HDL-C) in the mothers but not in offspring; high lard fat diet (HLF) increased serum total cholesterol level (TC) and low density lipoprotein cholesterol level (LDL-C) both in mothers and offspring. Brain fatty acids profile was altered by HLF compared with chow diet. Polyunsaturated fatty acids and long-chain polyunsaturated fatty acids content were significantly lower in the HLF group than in the control group, but saturated fatty acids content were significantly higher in HLF group than those in control group. The changed fatty acids composition affected the spatial memory and learning ability of adult offspring. CONCLUSIONS: A long-term high lard diet increased offspring serum TC and LDL-C levels and affected the brain's fatty acid composition, and memory and learning ability. The polyunsaturated fatty acid content of the brain may be correlated with serum cholesterol levels.