Neuroprotective Effect of Moxibustion on Cerebral Ischemia/Reperfusion Injury in Rats by Downregulating NR2B Expression

Objective. Stroke is a common and frequently occurring disease of the central nervous system, which is characterized by high mortality and a high disability rate. Moxibustion is a common method for treating stroke in traditional Chinese medicine, but its neuroprotective mechanism is unknown. N-Methyl-D-Aspartate Receptor Subunit 2B (NR2B) plays an important role in neuronal apoptosis. The objective of this study was to explore the mechanisms underlying the neuroprotective effect of moxibustion on cerebral ischemia/reperfusion (I/R) injury based on NR2B. Methods. Sprague–Dawley rats were randomly divided into 5 groups: the control group, I/R group, I/R + moxibustion group, I/R + Ro25-6981 (NR2B antagonist) group, and I/R + Ro25-6981 + moxibustion group. The cerebral ischemia/reperfusion model was induced by middle cerebral artery occlusion. Before the establishment of the model, the Ro25-6981 group received intraperitoneal injections of Ro25-6981, the moxibustion group received moxibustion, and the Ro25-6981 + moxibustion group received both interventions. The neurological dysfunction was evaluated by a neurological deficiency score (NDS). The infarct volume was examined by TTC (2,3,5-triphenyltetrazolium chloride) staining. The apoptosis rate of cerebral cells in the ischemic area was examined by TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling) staining, and the expression of Bcl-2, Bax, and caspase-3 was observed by western blot. NR2B and JNK were also observed by western blot. Results. Compared with the I/R group, moxibustion significantly decreased the neurological deficiency score ( P < 0.05) and the infarct rate ( P < 0.01) in I/R rats which were similar to those in the Ro25-6981 group. After moxibustion treatment, there was a significant decrease in the apoptosis rate ( P < 0.001) and the protein expression levels of Bax, caspase-3, and JNK ( P < 0.001) and an increase in the expression of Bcl-2 ( P < 0.01). Compared with the I/R group, moxibustion downregulated the expression of NR2B and decreased the activity of NR2B in the cerebral ischemia area ( P < 0.001). Conclusions. Moxibustion can improve neurological dysfunction and decrease infarction area and neuronal apoptosis caused by cerebral ischemia/reperfusion in rats. Its neuroprotective mechanism may be related to downregulating the expression of NR2B.

Dose-effect relationship of ginger interposed moxibustion for allergic rhinitis: study protocol for a randomised, placebo-controlled and parallel clinical trial

ABSTRACTIntroductionAllergic rhinitis has a severe impact on patients’ life quality, and the incidence rate keeps increasing. Moxibustion is widely used for treating allergic rhinitis, and quantity is the basis of moxibustion efficacy. The purpose of this study is to evaluate the relationship between the different quantities of moxibustion and the efficacy of moxibustion in the treatment of allergic rhinitis. This study may be conductive to the standardization of moxibustion and furnish the mechanism of dose-effect relationship of moxibustion with data and new ideas.Methods and analysisThis randomized, placebo-controlled clinical trial will include 33 patients with allergic rhinitis who will be randomly assigned into three groups in a 1:1:1 ratio: high-dose moxibustion group, low-dose moxibustion group and sham moxibustion control group. All groups will be treated once every other day, 20 days for one treatment course. And the patients will receive treatment for 2 courses with an interval of 2 days between courses. We will conduct a follow-up 30 days later after completion of treatments. The primary outcome measure is Total Nasal Symptom Score, carried out at baseline, 3, 6 and 10 weeks. Secondary outcome measure is Rhino Conjunctivitis Quality of Life Questionnaire, carried out at baseline, 6 and 10 weeks.Ethics and disseminationThis trail has been approved by the IRB of Shanghai TCM-Integrated Hospital, Shanghai University of TCM. The results of the trial will be disseminated in peer-reviewed journals.Trial registration numberChiCTR2100050373; Pre-results.Strengths and limitations of this studyA randomised, placebo-controlled and parallel clinical trial will be conducted to test if ginger interposed moxibustion would have curative effect on allergic rhinitis and reveal the preliminary dose-effect relationship of moxibustion.This study will set high-dose moxibustion group and low-dose moxibustion group to test if more quantities of ginger interposed moxibustion would achieve better therapeutic effects on allergic rhinitis.Further study should be carried out to test if the curative effect of ginger interposed moxibustion would be positively correlated with the quantities.

Regulation of Mild Moxibustion on Uterine Vascular and Prostaglandin Contents in Primary Dysmenorrhea Rat Model

Objective. Primary dysmenorrhea (PD) is a common and high incidence disease in gynecology, which seriously affects the quality of life in young women. Our previous study found that mild moxibustion could treat abdominal pain of PD patients, but the mechanism is still unclear. Therefore, this study aims to partly investigate the treatment mechanism of moxibustion for PD, especially on uterine microcirculation. Methods. Forty 3-month-old Sprague Dawley female rats were randomly divided into four groups, including group A (saline control group, n = 10), group B (control plus moxibustion group, n = 10), group C (PD model group, n = 10), group D (PD. model plus moxibustion group, n = 10). The PD rat model was established by injecting estradiol benzoate and oxytocin. Mild moxibustion on Sanyinjiao (SP6) and Guanyuan (CV4) acupoints was once a day, 20 minutes per time, for 10 consecutive days. A vaginal smear was used to test the estrous cycle of rats. Uterine microvascular thickness was observed by stereomicroscope. And we detected the content of prostaglandin F2α (PGF2α) and prostaglandin E2 (PGE2) in uterine tissue by enzyme-linked immunosorbent assay. Results. Mild moxibustion can enlarge the microvessels, improve the microcirculation disturbance, and relieve the swelling of the uterus in PD rats. During the mild moxibustion intervention, the contents of PGF2α and PGE2 in uterus issues were synchronous increases or decreases and the changes of PGE2 were more obvious, but the changes of uterine microvasculature and morphology caused by the decrease of PGF2α were greater than PGE2. Conclusion. Mild moxibustion at SP6 and CV4 acupoints may relax uterine microvascular obstacle by reducing the content of PGF2α in uterine tissue, improve the microcirculation disorder, and then alleviate the PD rat’s uterine swelling.

Effect of Thunder-Fire Moxibustion for Lumbar Disc Herniation: Study Protocol for a Randomized Controlled Trial

BackgroundLumbar disc herniation (LDH) is one of the common diseases in orthopedics and traumatology, which is caused by nucleus pulposus herniation stimulating cauda equina nerve, nerve root and intervertebral disc degeneration. Its clinical manifestations are low back pain, radiation pain of lower limbs and cauda equina symptoms, which seriously affect the quality of life of patients. At present, oral analgesics are commonly used in clinical non drug therapy for LDH, but oral analgesics will produce gastrointestinal reactions and other side effects. Thunder-fire moxibustion is one kind of moxibustion method, which has been applied widely for treating pain syndromes in China. The aim of our research is to design a randomized controlled trial of thunder-fire moxibustion in the treatment of lumbar disc herniation to explore whether it is safer and more effective than oral analgesic drugs.Methods90 patients will be randomly divided into thunder-fire moxibustion group and acetaminophen group. The intervention included 10 days as a course of treatment, lasting for 20 days. The acetaminophen group took two acetaminophen sustained-release tablets every other day, while the thunder fire moxibustion group will be treated with thunder fire moxibustion once every other time for 30 minutes. Japanese Orthopaedic Association (JOA) score, visual analogue scale (VAS) and oswestry disability index (ODI) will be used as the main the observation indexes. Meanwhile, the occurrence of adverse events (AES) will also be recorded. The assessment will be conducted at baseline and at the end of the first and second course of treatment.DiscussionThe aim of this study is to determine whether thunder-fire moxibustion is more effective than acetaminophen in the treatment of patients with LDH.Trial registrationChinese Clinical Trial Registry (http://www.chictr.org.cn), ChiCTR2000036079. Registered on 21 August 2020.

Dose-Effect of Long-Snake-Like Moxibustion for Chronic Fatigue Syndrome: Study Protocol for a Randomized Controlled Trial

Background Chronic fatigue syndrome (CFS) is a disease with high incidence rate and great impact on life, but it lacks for specific pharmacological treatment and diagnostic indicators. In the theory of traditional Chinese medicine (TCM), fatigue is the result of Yang deficiency. Long-snake-like moxibustion, as one of a special moxa therapy, has been applied in Yang deficiency patients for thousands of years in China and it is used widely to relieve fatigue symptoms for its strong function of Yang warming. However, the does-effect relationship is unclear in the moxibustion research. Hence, we design this trial to assess the duration-effect of long-snake-like moxibustion through combining measurements of the subjective patient-reported scales with objective medical infrared imaging technology─Thermal Texture Maps (TTM). Methods This is a single-center, randomized controlled trial. Thirty healthy women and sixty female CFS patients will be recruited to receive the first TTM scanning, then CFS patients will be allocated to 60-minute long-snake-like moxibustion (Group A) and 30-minute long-snake-like moxibustion (Group B) equally. These two groups will receive corresponding treatment once a day, three sessions per week every other day for consecutive 4 weeks. The second TTM scanning will be employed for CFS patients after the end of treatment. The primary outcome will be the score improvement of the Fatigue scale-14 (FS-14). Secondary outcomes include the change of the Self-rating depression scale (SDS),the Self-rating anxiety scale (SAS), and the Symptoms Scale of spleen-kidney Yang deficiency. Discussion The trial will provide evidence for the choice of treatment duration for long-snake-like moxibustion in treating CFS. The results will contribute to explore the dose-effect relationship of moxibustion, and optimize the efficacy of moxibustion therapy. Trial registration: Chinese Clinical Trail Registry (No. ChiCTR2000041000), on 16th December, 2020, http://www.chictr.org.cn/showproj.aspx?proj=62488

A Urinary Metabolomics Analysis Based on UPLC-MS and Effects of Moxibustion in APP/PS1 Mice

Background: Alzheimer's disease (AD) is a common neurodegenerative disorder with the symptoms of cognitive impairment and decreased learning and memory abilities. Metabolomics can reflect the related functional status and physiological and pathological changes in the process of AD. Moxibustion is a unique method in traditional Chinese medicine, which has been used in the treatment and prevention of diseases for thousands of years. Methods: A total of 32 APP/PS1 mice were randomly divided into the model group, moxibustion group, moxa smoke group and smoke-free moxibustion group (n=8/group), using the random number table method, while eight C57BL/6 mice were used as the control group. The five groups were measured for 20 min/day, 6 days/week, for 4 weeks. After 4 weeks’ experiment, all the mice were placed in metabolic cages to collect urine continuously for 24 hours, for UPLC-MS analysis. Results: Principal component analysis (PCA) was used to identify the different metabolites among the five groups, and partial least squares discriminant analysis (PLS-DA) was performed to reveal the effects on the metabolic variance. Sixteen potential biomarkers were identified among the five groups, primarily related to amino acid metabolism, starch metabolism, sucrose metabolism, interconversion of pentose and glucuronate, and aminoacyl biosynthesis. There were 17 differences in the potential metabolites between the control and model groups, involving the metabolism of amino acid, purine, pyrimidine, nicotinic acid and nicotinamide, and biosynthesis of pantothenate and coenzyme A. Fifteen potential biomarkers were identified between the model and moxibustion groups, related to starch metabolism, sucrose metabolism, interconversion of pentose and glucuronate, glyoxylate, dicarboxylate anions and some amino acid metabolism. Conclusion: Moxibustion can regulate the metabolism of substance and energy by improving the synthesis and decomposition of carbohydrates and amino acids in APP/PS1 transgenic AD model mice.

Effects of Moxibustion on Amygdala -HPA Axis in Rats with Kidney-Yang Deficiency Syndrome Induced by Hydrocortisone Injection

Background: The syndrome of kidney-yang deficiency is one of the main syndromes in traditional Chinese medicine. Modern evidences prove that the hypothalamus - pituitary - adrenal axis (HPA axis) function disorder is the main material basis of kidney-yang deficiency syndrome. The upper regulating center, such as hippocampus and amygdala can affected HPA axis. Although moxibustion have a therapeutic effect for kidney-yang deficiency syndromes, the underlying mechanism remains unclear. This study investigated the effect of suspended moxibustion on amygdala and HPA axis and elucidated the possible molecular mechanism of moxibustion in improving kidney-yang deficiency syndrome.Methods: 60 male Sprague-Dawley rats were randomly divided into the normal group ( n=12) and the model-building group (n=48). Rats in the model-building group were given intramuscular injection of hydrocortisone to establish the model of kidney-yang deficiency. The 48 rats successfully modeled were then randomly divided into model group (model, n=12) , carbenoxolone intraperitoneal injection group (CBX, n=12), moxibustion group (moxi, n=12) and moxibustion plus carbenoxolone intraperitoneal injection group (moxi+CBX, n=12). In the moxibustion group, Shenshu (BL23) and Guanyuan (CV4) points were treated with moxibustion for 14 days. After treatment, the level of corticostesone (CORT), adrenocorticotropic hormone (ACTH) and corticotropin-releasing hormone (CRH) in serum, the expressions of mineralocorticoid receptor (MR), glucocorticoid receptor (GR), 11 beta-hydroxysteroid dehydrogenase type 1 (11β-HSD1), corticotropin releasing factor (CRF) and ACTH in rats’ amygdala and (or) hypothalamus, pituitary were detected. Data were analyzed by one-way analysis of variance.Results: Compared with the normal group, the level of CRH, ACTH and CORT in serum, and the mRNA and protein expressions of MR, GR and 11β-HSD1in amygdala, the mRNA and protein expressions of 11β-HSD1 in hypothalamus and CRF mRNA expression in amygdala and hypothalamus, and ACTH mRNA expression in pituitary of rats in the model group were all significantly decreased (P < 0.05 or 0.01). After treatment with moxibustion, except for 11β-HSD1 mRNA expression, the observation index mentioned above were increased to different degrees compared with those in the model group (P < 0.05 or 0.01). Conclusion: Suspended moxibustion can effectively improve the serum levels of ACTH, CRH and CORT, and can up-regulate the mRNA and protein expression of MR, GR , 11β-HSD1, CRF and ACTH in amygdala and hypothalamus of kidney-yang deficiency rats, which may be one of the molecular mechanisms of moxibustion in improving kidney-yang deficiency syndrome.

Effect of Moxibustion on Behavioral Changes and Expression of APP and BACE1 in Hippocampus of SAMP8 Mice

Objective. To observe the effect of moxibustion on cognitive function of aging mice, to observe the effect of moxibustion on protein and gene expression of APP metabolism pathway, and to explore the mechanism of action in moxibustion. Methods. 24 SAMP8 were randomly divided into 2 groups (12 in each group): moxibustion group and model group. 12 SAMR1 mice were used as blank controls. Mice in the moxibustion group were treated with moxibustion for 8 weeks, 10 minutes each time, 5 times a week, and for a total of 8 weeks. The model group and the blank group were treated with sham-moxibustion. Behavior tests were used to detect the learning and memory ability of each group of mice. Immunohistochemical, western blot, and RT-PCR were used to detect the protein and mRNA expression of APP and BACE1. Furthermore, the expressions of miR-29 and miR-101 were observed by RT-PCR method to explore the mechanism of moxibustion at the genetic level. Results. In this study, relative to normal mice, we found that aging mice showed behavioral changes consistent with the onset of AD. However, moxibustion interventions were able to mitigate these effects to some degree in aging mice. In addition, moxibustion was proved to regulate APP metabolism pathway at protein and gene level through molecular biology tests. Conclusion. The data suggest that the effect of moxibustion intervention on cognitive function in aging mice is related to the regulation of genes and proteins involved in APP metabolism pathway; this may be a potential target for treating Alzheimer’s disease.

Observation of the effects of moxibustion on acute and chronic radiation enteritis in cervical patients

BackgroundThe present prospective, observational study evaluated the safety and efficacy of moxibustion in preventing acute and chronic radiation enteritis post-radiation in patients with cervical cancer.MethodsBetween March 2015 and October 2018, 90 patients with stage I-IV cervical cancer were treated with radiation at the Guangzhou University of Chinese Medicine Second Affiliated Hospital (Guangdong Provincial Hospital of Chinese Medicine). The patients were randomly divided into a moxibustion group and a control group. The moxibustion group received moxibustion on Guānyuán (RN4), Qìhǎi (RN6), Shénquē (RN8) and Tiānshū (ST25) during radiation. The control group received radiotherapy only. We observed and compared the occurrence of acute radiation enteritis (ARE) between the two groups of patients during radiotherapy. Then, the patients were followed-up 1-1.5-year after radiation, and we observed and compared the occurrence of chronic radiation enteritis (CRE) between the two groups.ResultsThe incidence of ARE was significantly lower in the moxibustion group (10.9%) than the control group (36.4%; P < 0.05). At follow-up post-radiotherapy, the incidence of CRE was significantly reduced (15.2%) in the moxibustion group compared with the control group (35.7%; P < 0.05).ConclusionsMoxibustion significantly improved physical function and reduce the gastrointestinal response caused by radiotherapy, thus the occurrence of acute and chronic radiation enteritis was reduced. These results can improve the therapeutic effect of radiation and improve the patient’s quality of life.Trial registrationProspective clinical trial for the effects of moxibustion on acute and chronic radiation enteritis in cervical patients. ChiCTR2000029650. Registered: February 9, 2020-Retrospectively registered.

Effect of Moxibustion on Testosterone Secretion and Apoptosis of Spermatogenic Cells in Aging Rats

Analysis of androgen secretion and sperm production was conducted in the testis to investigate the efficacy of moxibustion on testicular function in aging rats. Male Sprague–Dawley rats were randomly divided into the aging group (N = 8), the mild-warm moxibustion group (N = 8), and the youth control group (N = 8). Rats in the mild-warm moxibustion group (MWMG) were exposed to mild-warm moxibustion at the Zusanli (ST36) and Shenshu (BL23) acupuncture points daily, from the age of 12 months until the age of 24 months. After the intervention, testicular tissue was harvested from all rats across groups. Changes in testicular structure were examined by hematoxylin and eosin (H&E) stain. Detection of the apoptosis of spermatogenic cells was performed by the TUNEL assay. Testosterone level in the testis was analyzed by the ELISA assay, and the expression of Bax, Bcl-2, and androgen receptor (AR) in the testis was evaluated by immunohistochemistry. AR expression analysis was subsequently performed by the western blotting assay, and the detection of telomerase activity of the testis and the expression of Bax, Bcl-2, and AR mRNA were performed by real-time PCR. Compared with the youth controls, telomerase activity in the testis, testosterone levels, expression of AR, and expression of antiapoptosis factor Bcl-2 protein and mRNA were significantly decreased (P<0.01) in the aging group. Spermatogenic cell apoptosis (P<0.01) and proapoptotic factor Bax expression were significantly increased (P<0.01) in the aging rats compared with the youth control group. The MWMG exhibited significant increases in testicular telomerase activity, testosterone level, AR expression, and antiapoptosis factor Bcl-2 expression (P<0.05 or P<0.01) compared with the aging group. In this experimental group, spermatogenic apoptosis was inhibited (P<0.01) and proapoptotic factor Bax expression significantly reduced (P<0.01). Mild-warm moxibustion can inhibit reproductive senescence by improving telomerase activity, improving AR expression, restoring testosterone, and inhibiting spermatogenic apoptosis via regulation of Bcl-2/Bax.