Exploring the inhibitory impact of Mongolian medicinal He-Zi-3 soup on mammary gland hyperplasia in rats induced by estrogen and progestogen.

ETHNOPHARMACOLOGICAL RELEVANCE: Mammary gland hyperplasia, a prevalent benign breast condition, often serves as a precursor to various other breast diseases. He-Zi-3 soup (HZ-3), a traditional Mongolian remedy, is utilized for treating this condition. AIM OF THE STUDY: To explore the effect and underlying mechanism of HZ-3, a Mongolian medicinal preparation, on mammary gland hyperplasia. MATERIALS AND METHODS: This study aimed to assess the impact of different doses of HZ-3 in a rat model of mammary hyperplasia. The active components within HZ-3 drug serum were identified and analyzed through network pharmacology and target prediction. To elucidate the underlying mechanism of HZ-3 in addressing mammary hyperplasia, we conducted a series of investigations on estradiol-induced mammary hyperplasia in model rates. Assessments included measurements of papilla width and height, hematoxylin and eosin staining, Masson staining, reverse transcription-quantitative polymerase chain reaction (RT-qPCR), Western blot, and immunohistochemistry. RESULTS: Our investigation revealed the identification of 21 compounds, primarily terpenoids, through serum medicinal chemistry screening. Utilizing network pharmacological analysis, we observed predominant regulation through the estrogen pathway, closely associated with key genes including esr1,esr2, ncoa1, krt 19, ctsd, ebag 9, and bcl-2. Assessments encompassing nipple height and width, histological examination, immunohistochemical analysis, and serum hormone levels via enzyme-linked immunosorbent assay demonstrated the inhibitory effect of HZ-3 on mammary hyperplasia in rat models. RT-qPCR and Western blot analyses corroborated these findings, affirming the suppression of mammary hyperplasia by HZ-3 through the activation of estrogen pathway signaling.

Overview of Traditional Mongolian Medical Warm Acupuncture.

Mongolian medical warm acupuncture is a traditional therapy of Mongolian medicine and was developed by people living on the Mongolian Plateau. This kind of traditional oriental medicine has a long history. The main characteristics of Mongolian medical warm acupuncture are the acupoints and the needles used. Its theory is based on the human anatomical structure and the distinct local culture. Mongolian medical warm acupuncture has been practiced for centuries and proved to be very effective in the treatment of age-related diseases, including the musculoskeletal and nervous diseases. This paper aims to briefly introduce the history and scope of Mongolian medical warm acupuncture, with a particular focus on age-related diseases, where Mongolian medical warm acupuncture has shown significant beneficial effects.

[Effect of moxibustion on IL-6/STAT3 signaling in frontal cortex of fatigue rats].

OBJECTIVE: To observe the effect of moxibustion on interleukin-6(IL-6)/signal transduction and transcriptional activator 3(STAT3) signaling pathway in the frontal cortex of fatigue rats, so as to reveal its mechanisms underlying alleviation of fatigue. METHODS: Twenty-one male SD rats were randomly divided into normal control, model, and moxibustion groups (n＝7 rats in each group). The fatigue model was established by forcing the rats to have an exhausted swim under load condition, once daily for 21 days. Moxibustion was applied to bilateral "Zusanli"(ST36) for about 15 min, once every other day for 21 days. The level of IL-6 in the frontal cortex was detected by ELISA, and the expression of Janus kinase 2 (JAK2), phosphorylated JAK2 (p-JAK2), signal transduction and transcriptional activator 3(STAT3) and phosphorylated STAT3 (p-STAT3) proteins in the frontal cortex was detected by Western blot. RESULTS: After modeling, the levels of IL-6 content and p-STAT3 protein expression and ratio of p-STAT3/STAT3 were significantly increased in the model group relevant to the normal control group (P<0.01). Follo-wing moxibustion, the duration of load swimming on the 21st day was significantly prolonged (P<0.01), content of IL-6 and levels of p-STAT3 protein expression and ratio of p-STAT3/STAT3 were significantly down-regulated in the moxibustion group compared with the model group (P<0.05). No significant differences were found between the model and control, and between moxibustion and model groups in the expression levels of JAK2, p-JAK2, STAT3 and p-JAK2/JAK2 (P>0.05). CONCLUSION: Moxibustion intervention can relieve fatigue in fatigue rats, which is associated with its function in inhibiting IL-6/STAT3 signaling pathway to reduce inflammatory injury.

Moxibustion Reduces Inflammatory Response in the Hippocampus of a Chronic Exercise-Induced Fatigue Rat.

Accumulating data indicates that brain inflammation plays an important role in the pathophysiology of chronic exercise-induced fatigue. Moxibustion in traditional Chinese medicine has been found to alleviate exercise-induced fatigue. However, it remains unclear whether the effect of moxibustion is related to its anti-inflammatory properties. In this study, rats were exposed to 3-week exhaustive swimming to induce chronic exercise-induced fatigue. The body weight, exhaustive swimming time, tail suspension test and open-field test were observed. Real-time polymerase chain reaction (RT-PCR) was used to determine the mRNA expression of proinflammatory cytokines (interleukin-1ß [IL-1ß], interleukin-6 [IL-6], and tumor necrosis factor-α[TNF-α]), and enzyme-linked immunosorbent assay (ELISA) was used to detect IL-1ß, IL-6, and TNF-α concentrations. Chronic exhaustive exercise significantly reduced the body weight and exhaustive swimming time, and increased tail suspension immobility time, which were reversed by moxibustion treatment. Compared with control rats, the mRNA and protein expression of IL-1ß, IL-6, and TNF-α in the hippocampus was significantly increased in exhaustive swimming trained rats. Moxibustion significantly decreased the level of IL-6 in the hippocampus, but not affected IL-1ß and TNF-α level significantly. Our results suggested that a potential inflammatory damage in the brain may be involved during chronic exhaustive exercise-induced fatigue. Moxibustion could attenuate the inflammatory impairment in exercise-induced fatigue, which might be mediated by inhibition of the proinflammatory cytokine IL-6 levels in the brain region.

Effect of Mongolian warm acupuncture on the gene expression profile of rats with insomnia.

BACKGROUND: The mechanism of Mongolian warm acupuncture (MWA) for the treatment of insomnia has not been previously reported. OBJECTIVE: To investigate the effect of MWA on gene expression profile in the p-chlorophenylalanine (PCPA)-induced rat model of insomnia. METHODS: A rat model of insomnia was established and the animals were divided into five groups: control, PCPA (untreated), PCPA+estazolam, PCPA+MA (manual acupuncture), and PCPA+MWA. The rats were euthanased at 7 days after treatment, and hypothalamic tissue was harvested to extract total RNA for the analysis of gene expression profile. Micro-array and Partek Genomics Suite analysis system were used to analyse differential expression of genes between groups. Furthermore, ingenuity pathways analysis was used to analyse the main regulators. RESULTS: After treatment, in rats with improved sleep, micro-array data from the follow-up phase compared with baseline showed that MWA down-regulated 11 genes compared with the control group and 16 genes compared with the PCPA group. Six genes were selected following the micro-array detection to perform quantitative polymerase chain reaction (qPCR) verification, and the results showed that the coincidence rate was up to 90%, which verified the reliability of the microarray results. Compared with the PCPA group, transcription levels of Egr 1, Btg2 and BDNF in the PCPA+MWA group were up-regulated (P<0.05). CONCLUSION: In combination, the findings of this study suggests that MWA is efficacious at improving sleep in an experimental rat model of insomnia.

[Effect of moxibustion on inflammatory cytokine levels in hippocampus and hypothalamus in rats with fatigue].

OBJECTIVE: To investigate the effect of moxibustion of "Zusanli "(ST36) on levels of inflammatory cytokines in the hippocampus and hypothalamus regions of rats with fatigue, so as to reveal its anti-fatigue mechanism. METHODS: Twenty-four male Sprague-Dawley rats were randomly divided into normal control, fatigue model, and moxibustion groups, with 8 rats in each group. The fatigue model was established by chronic weight-loaded exhaustive swimming. During modeling, the rats in the moxibustion group were given moxibustion (3 moxa cones, about 8 min in duration) at bilateral ST36, once every other day for 11 times in total. The duration of exhausted swimming was observed, and ELISA was used to measure the contents of interleukin-1ß (IL-1ß), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and interferon gamma (IFN-γ) in the hippocampus and hypothalamus tissues. RESULTS: The duration of exhausted swimming was obviously prolonged on day 14 and 21 after moxibustion intervention relevant to the model group (P<0.05, P<0.01). The contents of IL-1ß， IL-6 and TNF-α in the hippocampus were significantly increased in the model group than in the normal group (P<0.01), and that of hippocampal IL-6 was considerably decreased in the moxibustion group than in the model group (P<0.05). The content of hippocampal IFN-γ was significantly lower in the model group than in the normal group (P<0.05) and was increased mildly after moxibustion (P>0.05). No significantly changes were found in the levels of hypothalamic IL-1ß， IL-6, TNF-α and IFN-γ after modeling and moxibustion (P>0.05), and in the levels of hippocampal IL-1ß， TNF-α and IFN-γ after moxibustion (P>0.05)．. CONCLUSION: Moxibustion can significantly down-regulate hippocampal IL-6 content in fatigue rats, which maybe contribute to its effect in alleviating fatigue. Further studies need being conducted to identify this conclusion.

Effects of Mongolian Warm Acupuncture on iNOS/NO and Inflammatory Cytokines in the Hippocampus of Chronic Fatigue Rats.

The inducible nitric oxide synthase/nitric oxide (iNOS/NO) signaling pathway and inflammatory cytokines play important roles in the pathogenesis of exercise-induced fatigue. Studies have found that Mongolian warm acupuncture (WA) could alleviate exercise-induced fatigue. However, the exact mechanisms underlying its effects remain unclear. In the present study, we investigated the effects of Mongolian WA on iNOS/NO signaling pathway and proinflammatory cytokines in a chronic exhaustive swimming-induced fatigue rat model. Animals were randomly divided into Control group, Ctrl + WA group, Model group, and Model + WA group. The body weight, exhaustive swimming time test, and Morris water maze test were performed before and after the chronic exhaustive swimming. The serum levels of interleukin-1ß (IL-1ß), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), and iNOS were detected by enzyme linked immunosorbent assay (ELISA). The mRNA expressions of IL-1ß, IL-6, TNF-α, IFN-γ, and iNOS in the hippocampus were measured by real-time polymerase chain reaction (RT-PCR). Moreover, the protein expression of iNOS in the hippocampus was measured by western blot, and the NO productions in the serum and hippocampus were detected by Griess reaction system. Chronic exhaustive exercise significantly reduced the body weight and exhaustive swimming time, and induced impairment in learning and memory, and which were reversed by WA treatment. Chronic exhaustive exercise also increased the expressions of iNOS and proinflammatory cytokines, while WA treatment significantly decreased the level of iNOS and proinflammatory cytokines. However, chronic exhaustive exercise did not affect the NO production. These findings demonstrated that WA could alleviate the chronic exhaustive swimming-induced fatigue and improve the learning and memory ability, and the actions might be related to the reduction of inflammatory response and iNOS expression.

[Effects of electro-warmed needle of inner-Mongolian medicine on serum TNF-alpha, ACTH and corticosterone contents in fatigue rats].

OBJECTIVE: To observe the effect of electro-warmed needle (EWN, of Inner-Mongolian medicine) on fatigue rats' behavior, hypothalamic-pituitary-adrenocortex (HPA) axis activity and immune system so as to reveal its neuro-endocrino-immune mechanism. METHODS: Male SD rats were randomized into control (n=20), model (n=20) and EWN (n=19) groups. Fatigue model was established by forcing the rat to swim in a water pool till exhaustion, once daily, continuously for 21 days. "Dinghui" (central spot over the bregmatic bone) and "Xinxue" (the center of the depression beneath the 7th thoracic vertebra) were punctured with silver needles which were warmed electrically by using a MLY-I Electrical Needle-warming Apparatus, once every 3 days, 7 sessions altogether. On the 21st day of modeling, swim-exhaustion duration (SED), and immobility time and struggle times in tail suspension test were measured. Twenty-four hours after the last swim, the rats' serum TNF-alpha, ACTH an corticosterone contents were detected by enzyme linked immunosorbent assay (ELISA). RESULTS: Compared with control group SED, immobility time and struggle times in tail suspension test in model group decreased, increased and lowered respectively and significantly (P < 0.01, 0.05); while in comparison with model group, the first 2 indexes of EWN group increased and lowered respectively and significantly (P < 0.05). No significant difference was found between EWN and model groups in struggle times (P > 0.05). Compared with control group, serum TNF-alpha, ACTH and corticosterone contents in model group increased significantly (P < 0.01), while in comparison with model group, the 3 indexes of EWN group were significantly lower (P < 0.01, 0.05). CONCLUSION: EWN treatment can reduce fatigue-induced increase of serum TNF-alpha, ACTH and corticosterone levels, and raise motor ability, suggesting a favorable regulation of HPA axis and immune function after EWN and improvement of fatigue in fatigue

[Effects of electric mildly-warmed needle of inner mongolian medicine on liver MDA and GSH content, GSH-Px and SOD activity in fatigue rats].

OBJECTIVE: To investigate the effect of electric mildly-warmed needle of inner Mongolian medicine on changes of free radical metabolism in the liver tissue of rats with fatigue. METHODS: A total of 33 male SD rats were evenly randomized into control, model and electro-mildly-warmed needle (EMWN) groups. Fatigue model was established by forcing the rat to swim in a water pool till exhaustion, once daily, continuously for 21 days. "Dinghui" (central spot of the bregmatic bone) and "Xinxue" (the center of the depression beneath the 7th thoracic vertebra) were punctured with silver needle which was warmed electrically by using a MLY-I Electrical Needle-warming Apparatus, once every 3 days. 7 sessions altogether. After decapitation, the rat's liver was taken, homogenated and centrifuged for detecting malondialdehyde (MDA) content with thio-barbituric acid, reduced glutathione (GSH) contents, glutathione peroxidase (GSH-Px) activity with chromatometry and superoxide dismutase (SOD) activity with xanthine oxidase method respectively. RESULTS: Compared with control group, liver GSH content, GSH-Px and SOD activity in model group lowered significantly (P < 0.05, 0.01), while in comparison with model group, these 3 indexes of EMWN group increased considerably (P < 0.05, 0.01). No significant differences were found among 3 groups in MDA levels (P > 0.05). The results showed that the effect of mildly-warmed needle treatment could resist fatigue induced decline of activity of antioxidant enzymes. CONCLUSION: Electric mildly-warmed needle therapy of inner Mongolian medicine can raise the activity of antioxidant enzymes in liver tissue of fatigue rats, which may underlie the action of mildly-warmed needle in improving fatigue.