Effect of moxibustion on the SIRT1/FOXO3a signaling pathway in ApoE-/- mice with atherosclerosis. / 艾灸对ApoE-/-åŠ¨è„‰ç²¥æ ·ç¡¬åŒ–å°é¼ SIRT1/FOXO3a信号通路的影响.

OBJECTIVES: To observe the effects of moxibustion on blood lipid metabolism, pathological morphology of thoracic aorta, and the expression of silent information regulator 1 (SIRT1) and forkhead box transcription factor O3a (FOXO3a) in ApoE-/- atherosclerosis (AS) mice, so as to explore the potential mechanism of moxibustion in preventing and treating AS. METHODS: Ten C57BL/6J mice were fed a normal diet as the control group, and 30 ApoE-/- mice were fed a high-fat diet to establish the AS model, which were randomly divided into the model group, simvastatin group, and moxibustion group, with 10 mice in each group. From the first day of modeling, mice in the moxibustion group received mild moxibustion treatment at "Shenque"(CV8), "Yinlingquan"(SP9), bilateral "Neiguan"(PC6) and "Xuehai"(SP10) for 30 min per time；the mice in the simvastatin group were given simvastatin orally (2.5 mg·kg-1·d-1), with both treatments given once daily, 5 times a week, with a total intervention period of 12 weeks. The body weight and general condition of the mice were observed and recorded during the intervention period. After the intervention, the contents of serum total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) were measured using an automated biochemistry analyzer. Hematoxylin eosin (HE) staining was used to observe the pathological morphology of the thoracic aorta. ELISA was used to measure the contents of serum oxidized low-density lipoprotein (ox-LDL) and superoxide dismutase (SOD) activity. Western blot and real-time fluorescent quantitative PCR analysis were used to detect the expression levels of SIRT1 and FOXO3a protein and mRNA in the thoracic aorta. RESULTS: Compared with the control group, body weight at the 8th and 12th week, serum TC, TG, LDL-C, and ox-LDL contents of the model group mice were significantly increased(P<0.05, P<0.01), while the HDL-C contents, SOD activity, and the expression levels of SIRT1 protein and mRNA in the thoracic aorta were significantly decreased(P<0.05, P<0.01). HE staining showed thickening of the aortic intima, endothelial cell degeneration, swelling, and shedding. Compared with the model group, body weight at the 8th and 12th week, serum TC, TG, LDL-C, and ox-LDL contents of mice in the simvastatin group and moxibustion group were significantly decreased(P<0.01), while the serum SOD activity, expression levels of SIRT1 protein and mRNA in the thoracic aorta were significantly increased(P<0.01). The HDL-C contents were significantly increased in the simvastatin group(P<0.05). The thoracic aortic structure was more intact in both groups, with a more regular lumen and orderly arrangement of the elastic membrane in the media, and a slight amount of endothelial cell degeneration and swelling in the intima. There was no significant difference in the evaluated indexes between the moxibustion group and the simvastatin group and the pathological changes in the thoracic aorta were similar between the two groups. CONCLUSIONS: Moxibustion can reduce the body weight of AS model mice, regulate lipid levels, repair vascular intima, and alleviate endothelial damage. Its mechanism of action may be related to the regulation of the SIRT1/FOXO3a signaling pathway to improve oxidative damage.

Effect of acupoint catgut embedding on p38 MAPK pathway in the lung tissue of asthmatic rats. / 穴位埋线通过调节p38ä¸è£‚åŽŸæ´»åŒ–è›‹ç™½æ¿€é ¶é€šè·¯å¯¹å“®å–˜å¤§é¼ æ°”é“ä¸Šçš®çš„å½±å“.

OBJECTIVES: To observe the effect of catgut embedding at "Feishu"(BL13), "Dingchuan" (EX-B1) and "Danzhong" (CV17) on expression of phosphorylated p38 mitogen activated protein kinase (p-p38 MAPK), interleukin-4 (IL-4), interferon-γ (IFN-γ) and changes of airway epithelial cells (AEC) in the lung tissue of bronchial asthma (BA) rats, so as to explore its mechanisms underlying improvement of BA. METHODS: Forty male Wistar rats were randomly and equally divided into blank control, model, dexamethasone (DEX) and catgut embedding groups. The BA model was established by intraperitoneal injection of suspension of ovalbumin and aluminum hydroxide. Rats of the DEX group received intraperitoneal injection of DEX (1.5 mg/kg), once daily for 2 weeks, and those of the catgut embedding group received catgut embedding at BL13, EX-B1 and CV17 only one time. The rats' sneezing times per miniute in each group were recorded. H.E. staining was used to observe the histopathological changes of the lung tissue under light microscope. A transmission electron microscope (TEM) was used to observe the ultrastructural changes of AEC in the lung tissue, including the thickness of bronchial wall and bronchial smooth muscle by using an image analysis software. The protein expressions of p-p38 MAPK, IL-4 and INF-γ in the lung tissue were determined using Western blot. RESULTS: Morphological observation revealed that in the model group, light microscope showed deformed and swollen bronchial tube wall with increased folds and thickened bronchial smooth muscle；and TEM showed a large number of autophagy vesicles containing swollen and deformed organelles in the AEC, and apparent reduction of intracellular mitochondria, these situations were obviously milder in both DEX and catgut embedding groups. Compared with the blank control group, the sneezing times, thickness of bronchial wall and bronchial smooth muscle in the model group were significantly increased (P<0.01), and the expressions of p-p38 MAPK and IL-4 in lung tissue were significantly increased (P<0.01), while the expression of IFN-γ was significantly decreased (P<0.01) in the model group. In comparison with the model group, the sneezing times, thickness of bronchial wall and bronchial smooth muscle, protein expressions of p-p38 MAPK and IL-4 were significantly decreased (P<0.01), while the expression of IFN-γ was obviously increased (P<0.01) in both the DEX and catgut embedding groups. CONCLUSIONS: Acupoint catgut embedding can reduce the expression of IL-4 and increase the expression of IFN-γ by inhibiting p38 MAPK signal pathway of lung tissues in BA rats, which may contribute to its effect in alleviating the degree of airway epithelial cells damage.

Effect of acupuncture on the expression of neuropeptides and related inflammatory factors in rats with diarrhea-predominant irritable bowel syndrome. / é’ˆåˆºå¯¹è ¹æ³»åž‹è‚ æ˜“æ¿€ç»¼åˆå¾å¤§é¼ è„‘è‚ è‚½åŠç›¸å ³ç‚Žæ€§å› å­è¡¨è¾¾çš„å½±å“.

OBJECTIVES: To observe the effect of acupuncture on the expressions of neuropeptides and related inflammatory factors in rats with diarrhea-predominant irritable bowel syndrome(IBS-D), so as to explore the mechanism of acupuncture in the treatment of IBS-D. METHODS: Male Wistar rats were randomly divided into blank group, model group, medication group, and acupuncture group, with 6 rats in each group. Except for the blank group, the other groups were subjected to 14-day "acetic acid enema + restraint stress" to establish the IBS-D rat model. After successful modeling, the medication group received gavage of pinaverium bromide(15 mg/kg) once a day, and the acupuncture group received acupuncture at "Baihui"(GV20) and bilateral "Tianshu"(ST25), "Shangjuxu"(ST37), "Zusanli"(ST36), and "Taichong"(LR3) for 20 min every day, both groups were treated continuously for 14 days. The general state of the rats in each group was observed, and the body weight of the rats was measured. The open-field experiment was conducted to measure the horizontal and vertical movements, and the number of fecal pellets of rats. The histopathological morphology of hypothalamus and colon of rats was observed by HE staining. Toluidine blue staining was used to observe and count the mast cells(MCs) in the colon tissue of rats. ELISA was used to detect the serum contents of tumor necrosis factor-α(TNF-α) and interleukin(IL)-10. Real-time fluorescence quantitative PCR was performed to detect the mRNA expressions of calcitonin gene-related peptide(CGRP) in the hypothalamus and colon tissue. Western blot was used to detect the expressions of corticotropin-releasing factor(CRF) in the hypothalamus and colon tissue. RESULTS: HE staining showed that there was inflammatory cell infiltration in the lamina propria of colon in the model group, and it was reduced in the other groups. Compared with the blank group, the model group showed significantly decreased body weight, decreased walking distance and upright times in open field experiment, decreased serum IL-10 contents(P<0.05, P<0.01), increased fecal pellet number (P<0.01), increased MC number in the colon tissue, serum TNF-α contents, and CGRP mRNA expressions and CRF expressions in the hypothalamus and colon tissue(P<0.01). Compared with the model group, both medication and acupuncture groups showed significantly increased body weight, walking distance and upright times in the open-field experiment, and serum IL-10 contents(P<0.01, P<0.05), significantly decreased fecal pellet number (P<0.05), significantly decreased MC number in the colon tissue, serum TNF-α contents, and CGRP mRNA expressions in the hypothalamus and colon tissue(P<0.01)；at the same time, the acupuncture group showed significantly decreased CRF expressions in the hypothalamus and colon tissue(P<0.01, P<0.05). There was no significant difference in the above indicators between the medication group and the acupuncture group. CONCLUSIONS: Acupuncture can improve the general and emotional state, inflammatory response, and neuropeptide expression in rats with IBS-D, and alleviate the symptoms of IBS-D, which may be related to the regulation of neuropeptides and inflammatory factors levels.

UPLC-Q TOF-MS-Based metabolomics and anti-myocardial ischemia activity of Dioscoreae Nipponicae Rhizoma from different geographical origins.

The dried rhizome of Dioscorea nipponica Makino ("Chuanshanlong" in Chinese) is a medicinal herb with multiple major producing areas. The main objective of this study was the comparative profiling of Dioscoreae Nipponicae Rhizoma (DNR) from various geographical origins. A hypoxia/reoxygenation-induced H9c2 cell injury model was established, and the antimyocardial ischemia activity of DNR samples from different origins was detected using the cell counting kit-8 (CCK-8) method. The result showed that the antimyocardial ischemia potential of DNR samples from the Heilongjiang province was higher than that of the other studied samples. Subsequently, a plant metabolomics technique utilizing ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q TOF-MS) was used to determine the differences in DNR samples from various geographical origins. Forty compounds, including steroidal saponins, free fatty acids, and organic acids, were tentatively identified based on UPLC-Q TOF-MS fragmentation pathways and via comparison with available reference standards. Partial least squares discriminant analysis was performed to estimate the differences in DNR samples from different origins. Five compounds were significantly up-regulated and correlated with antimyocardial ischemia in DNR samples from Heilongjiang province. Molecular docking was used to discern the interactions of key markers with the active sites of the target protein. The findings signified that UPLC-Q TOF-MS metabolomics coupled with molecular docking is a powerful tool to rapidly identify the quality control characteristics of DNR samples and their products. The research provides a direction for the rational utilization of DNR.

[Effect of mild moxibustion on SIRT1/NF-κB signaling in atherosclerotic rabbits].

OBJECTIVE: To observe the effect of mild moxibustion on blood lipid, histopathological structure of the aortic arch, thoracic aortic silent information regulator 1 (SIRT1)/nuclear factor κB (NF-κB) signaling pathway in atherosclerosis (AS) rabbits, so as to explore its underlying mechanisms in improving AS. METHODS: Sixty male rabbits were randomly divided into control group (n=12), model group(n=11), mild moxibustion group (n=11), mild moxibustion + blocker (blocker) group (n=12). The AS model was established by feeding the rabbits with high-fat forage for 8 weeks, followed by immune response damage. Mild moxibustion was applied to "Danzhong"(CV17), "Shenque"(CV8) and "Neiguan" (PC6, bilateral) and "Xuehai" (SP10, bilateral) for 30 min, once daily, 3 times a week for 4 weeks. Rabbits of the blocker group received intraperitoneal injection of EX527 (a selective inhibitor of SIRT1, 5 mg·kg-1·d-1) 30 min before moxibustion. Rabbits of the control and model groups were only grabbed and fixed without intervention. After the intervention, the contents of serum triglyceride (TG) and total cholesterol (TC) were determined by enzymatic method, and those of serum low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) were determined by colorimetric method. The Sudan Ⅳ staining was employed to observe the histopathological structure of the aortic arch, and Western blot and fluorescence quantitative real time-PCR were used to detect the expressions of SIRT1 and NF-κB proteins and mRNAs in the thoracic aorta, respectively. RESULTS: Compared with the control group, the contents of serum TG, TC and LDL-C and the expression levels of NF-κB protein and mRNA were significantly increased (P<0.01, P<0.05), whereas the content of HDL-C and the expression of SIRT1 mRNA markedly decreased in the model group (P<0.01). After mild moxibustion, the contents of serum TG, TC, and LDL-C and the expression of NF-κB protein and mRNA were significantly down-regulated (P<0.01, P<0.05), while the content of HDL-C and the expression levels of SIRT1 protein and mRNA significantly up-regulated in the mild moxibustion group (P<0.05, P<0.01). There were no significant differences between the blocker and model groups in all the indexes (P>0.05). Compared with the mild moxibustion group, the serum TG, TC, and LDL-C contents and NF-κB protein expression were significantly increased (P<0.01, P<0.05), and HDL-C content and the expression of SIRT1 protein and mRNA significantly decreased (P<0.05, P<0.01) in the blocker group. Sudan Ⅳ staining showed vague structure of the aortic arch with obvious lipid infiltration in the model group, which was relatively milder in the mild moxibustion. CONCLUSION: Mild-moxibustion can reduce blood lipid levels and endothelial damage in atherosclerotic rabbits, which may be related to its function in regulating SIRT1/NF-κB signaling pathway.

Herbal therapy for ameliorating nonalcoholic fatty liver disease via rebuilding the intestinal microecology.

The worldwide prevalence of nonalcoholic fatty liver disease (NAFLD) is increasing, and this metabolic disorder has been recognized as a severe threat to human health. A variety of chemical drugs have been approved for treating NAFLD, however, they always has serious side effects. Chinese herbal medicines (CHMs) have been widely used for preventing and treating a range of metabolic diseases with satisfactory safety and effective performance in clinical treatment of NAFLD. Recent studies indicated that imbanlance of the intestinal microbiota was closely associated with the occurrence and development of NAFLD, thus, the intestinal microbiota has been recognized as a promising target for treatment of NAFLD. In recent decades, a variety of CHMs have been reported to effectively prevent or treat NAFLD by modulating intestinal microbiota to further interfer the gut-liver axis. In this review, recent advances in CHMs for the treatment of NAFLD via rebuilding the intestinal microecology were systematically reviewed. The key roles of CHMs in the regulation of gut microbiota and the gut-liver axis along with their mechanisms (such as modulating intestinal permeability, reducing the inflammatory response, protecting liver cells, improving lipid metabolism, and modulating nuclear receptors), were well summarized. All the knowledge and information presented here will be very helpful for researchers to better understand the applications and mechanisms of CHMs for treatment of NAFLD.

Clinical efficacy evaluation of a traditional Miao technique of crossbow needle therapy in the treatment of knee osteoarthritis: a multi-center randomized controlled trial.

BACKGROUND: Knee osteoarthritis (KOA) seriously reduces quality of life and is a major threat to the health of the middle-aged and elderly. This study aimed to assess the efficacy of Miao crossbow needle therapy vs. acupuncture for KOA therapy. METHODS: This multicenter, randomized controlled trial was performed at three hospitals between April 2016 and December 2016. The patients were randomized to receive crossbow needle (CN) or acupuncture (AT). All treatments were completed within 46 days. Evaluation of treatment was conducted on the 46th, 62nd, and 77th days. The primary endpoint was change of Western Ontario and McMaster Osteoarthritis Index (WOMAC) score on the 46th day. The secondary endpoints included WOMAC score, the Lysholm knee score, the Japanese Orthopedic Association (JOA) knee score, visual analog scale (VAS), and the MOS 36-item short-form health survey (SF-36), on the 46th, 62nd, and 77th day. RESULTS: Finally, data of 301 participants were analyzed for the efficacy of treatment. Compared with AT, there was a larger change of WOMAC score in the CN group after treatment [- 25.0 (95% CI - 27.0, - 23.0) vs. - 18.8 (95% CI - 20.8, - 16.9), P < 0.001]. In the CN group, the WOMAC score was lower at all three time points (P = 0.008, P = 0.003, P < 0.001 respectively), while the Lysholm knee score (P = 0.03) and JOA score (P = 0.013) were higher and the VAS score (P = 0.011) was lower on the 77th day. CONCLUSION: Both Miao crossbow needle therapy and acupuncture reduced the WOMAC score. Miao crossbow needle therapy can be an alternative method for treating patients with knee osteoarthritis. TRIAL REGISTRATION: ChiCTR, ChiCTR-INR-16008032. Registered on 12 March 2016.

[Herbal-cake-partitioned moxibustion of "Shenque" (CV8) has a relative specific effect in relieving abdominal pain and in regulating neuroendocrine-immune network in primary dysmenorrhea rats].

OBJECTIVE: To observe the effect of herbal-cake-partitioned moxibustion (HCPM) of "Shenque" (CV8) and "Daheng" (SP15) on abdominal pain, plasma ß-endorphin (ß-EP), uterine prostaglandin E2 (PGE2) and prostaglandin F2α (PGF2α) levels, as well as splenetic natural killer cell (NK cell) activity in primary dysmenorrhea (PD) rats, so as to explore the specificity of acupoint function and the underlying mechanisms of moxibustion in relieving dysmenorrhea. METHODS: A total of 40 female rats were randomized into blank control, model, CV8-direct moxibustion, CV8-HCPM and SP15-HCPM groups (n＝8 rats in each). The PD model was established by subcutaneous injection of estradiol benzoate injection (0.2-0.5 mg/rat) for 10 consecutive days and intraperitoneal injection of oxytocin (2 U) 24 h after the last subcutaneous injection. Moxibustion or herbal-cake (composed of Radix Angelicae Sinensis, Rhizoma Chuanxiong, Radix Paeoniae Rubra, Cortex Cinnamomi, etc.)-partitioned moxibustion was applied to CV8, SP15 or umbilicus respectively for 7 moxa-cones every time, once daily for 10 successive days. The rats of the control and model groups were also restrained as those in the moxibustion groups. The writhing times within 30 minutes was recorded and the contents of plasma ß-EP, uterine PGE2 and PGF2α were detected by ELISA, and NK cell activity was detected using MTT. RESULTS: Compared with the control group, the writhing times and the content of PGF2α in the uterus tissue were significantly increased in the model group (P<0.01), while the contents of plasma ß-EP, uterine PGE2 and splenetic NK cell activity were significantly decreased (P<0.01). In comparison with the model group, the writhing times and uterine PGF2α content were obviously down-regulated in the SP15-HCPM, CV8-direct moxibustion and CV8-HCPM groups (P<0.05, P<0.01), and the contents of plasma ß-EP and uterine PGE2, and splenetic NK cell activity were significantly increased (P<0.05, P<0.01). The therapeutic effects of CV8-HCPM group were significantly superior to those of SP15-HCPM and CV8-direct moxibustion groups in lowering writhing times and PGF2α level, and in up-regulating ß-EP, PGE2 (P<0.05, P<0.01). The NK cell activity of CV8-HCPM group was significantly increased compared with that in the SP15-HCPM group(P<0.05). No significant differences were found between the SP15-HCPM and CV8-direct moxibustion groups in the levels of writhing times, plasma ß-EP and uterine PGE2, PGF2α contents and splenetic NK cell activity (P>0.05). CONCLUSION: Moxibustion of both CV8 and SP15 can relieve abdominal pain in PD rats, which may be closely associated with its effect in suppressing PD-induced decrease of plasma ß-EP and uterine PGE2 levels and splenetic NK cell activity and increase of uterine PGF2α. The therapeutic effect of CV8-HCPM is obviously better than that of SP15-HCPM and CV8-direct moxibustion.

[Effects of the Acupoint Catgut Embedding on Nerve-Endocrine-Immune Network in Dysmenorrhea Rats].

OBJECTIVE: To explore the underlying mechanism of acupoint catgut embedding in improving primary dysme-norrhea (PD) in rats based on functional activities of the neuro-endocrine-immune (NEI) network. METHODS: Forty female rats were equally randomized into blank control, PD model, medication, and acupoint catgut embedding groups. The PD model was established by subcutaneous injection of estradiol benzoate (0.5 mg/rat on the 1st and 10th d, and 0.2 mg/rat from 2nd to 9th d) and oxytocin (2 U/rat, i．p.). Rats of the medication group were treated by intragastric perfusion of fenbid (0.8 mL/rat, 125 mg/100 mL), once daily for 10 days. The catgut embedding was applied to bilateral "Ciliao" (BL 32), "Sanyinjiao" (SP 6) and "Guanyuan" (CV 4) before modeling. The body writhing times in 30 minutes were recorded, plasma ß-endorphin(ß-EP) content, and prostaglandin E 2 (PGE2) and prostaglandin F 2 α (PGF2α) contents in the uterus tissue were assayed using ELISA, and the activity of natural killer cell (NK cell) in the spleen tissue was detected using 3-(4，5-dimethyl-2-thiazolyl)-2，5-diphenyl-2-H-tetrazolium bromide (MTT) method after isolation and co-culture with K 562 cells. RESULTS: The body writhing times were no-tably more in the model group than in the control group (P<0.01), and obviously fewer in both medication and catgut embedding groups than in the model group (P<0.01). After modeling, the plasma ß-EP and uterus PGE2 contents and splenic NK cell activity were significantly decreased (P<0.01), while the uterus PGF2α content was evidently increased in the model group relevant to the control group (P<0.01). Following the treatment, plasma ß-EP and uterus PGE2 contents and splenic NK cell activity were considerably up-regulated (P<0.01), and uterus PGF2α content was markedly down-regulated in both medication and acupoint catgut embedding groups (P<0.01), suggesting an involvement of the NEI network in catgut embedding-induced improvement of PD. The therapeutic effect of catgut embedment was markedly superior to that of medication in up-regulating splenic NK cell activity (P<0.01). No significant differences were found between the medication and catgut embedding groups in the body writhing times within 30 min, and in the levels of plasma ß-EP and uterus PGE2 and PGF2α (P>0.05). CONCLUSION: The acupoint catgut embedding has a significant efficacy in relieving PD in rats, which may be related to its effect in up-regulating plasma ß-EP, uterus PGE2 contents and splenic NK cell activity and in down-regulating uterus PGF2α level.

[Effect of medicinal-cake-separated moxibustion on functional activity of back-leg and plasma substance P level in patients with lumbar disc herniation].

OBJECTIVE: To observe the effect of medicinal-cake-separated moxibustion combined with acupuncture on back-leg activities and plasma substance P (SP) levels in patients with lumbar disc herniation, so as to reveal its mechanism underlying pain relief. METHODS: A total of 114 patients with lumbar disc herniation were randomly divided into control group (n=56) and treatment group (n=58) according to a random digits table. Patients of the control group were treated by manual acupuncture stimulation of main acupoints Jiaji (EX-B 2), Huantiao (GB 30, affected side), Chengshan (BL 57, affected side), Kunlun (BL 60, affected side), and supplemented acupoints Yanglingquan (GB 34), Weizhong (BL 40) and Zusanli (ST 36) in combination with wheat-flour-cake separated moxibustion at the main acupoints, and patients of the treatment group were treated by medicinal-cake [Chuanwu (Radix Aconiti), Caowu (Radix Aconiti Kusnezoffii), Ruxiang (Olibanum), etc. ]-separated moxibustion in combination with manual acupuncture stimulation of the same acupoints mentioned above. Acupuncture treatment was conducted for 30 min, followed by moxibustion for 15 min. The treatment was given once daily for 10 days. The patients' back-leg functional activity ability was assessed using straight-leg raising test, and the pain state assessed using visual analogue scale (VAS) and Japanese Orthopaedic Association (JOA) scores, respectively. The therapeutic effect was evaluated by using "Crite- ria for Diagnosis and Outcome Evaluation of Clinical Disorders or Syndromes of Chinese Medicine" issued in 1994 and plasma SP content was detected by radioimmunoassay. RESULTS: After the therapy, the back-leg activity score and JOA score of both groups were significantly higher than those of pre-treatment in the same one group (P<0. 05, P<0. 01), and those of the treatment group were significantly higher than those of the control group (P<0.05). The VAS score of the treatment group was re- markably lower than that of the control group (P<0. 01). After the treatment, the plasma SP content was markedly lower in the treatment group than in the control group (.P<0O. 05 ) . CONCLUSION: Medicinal-cake-separated moxibustion therapy can ame- liorate pain severity and functional activity of the back-leg pain patients with lumbar disc hernia, which may be related to its effect in reducing blood SP level.

Curcumin serves as a human kv1.3 blocker to inhibit effector memory T lymphocyte activities.

Curcumin, the principal active component of turmeric, has long been used to treat various diseases in India and China. Recent studies show that curcumin can serve as a therapeutic agent for autoimmune diseases via a variety of mechanisms. Effector memory T cells (T(EM), CCR7⁻ CD45RO⁺ T lymphocyte) have been demonstrated to play a crucial role in the pathogenesis of T cell-mediated autoimmune diseases, such as multiple sclerosis (MS) or rheumatoid arthritis (RA). Kv1.3 channels are predominantly expressed in T(EM) cells and control T(EM) activities. In the present study, we examined the effect of curcumin on human Kv1.3 (hKv1.3) channels stably expressed in HEK-293 cells and its ability to inhibit proliferation and cytokine secretion of T(EM) cells isolated from patients with MS or RA. Curcumin exhibited a direct blockage of hKv1.3 channels in a time-dependent and concentration-dependent manner. Moreover, the activation curve was shifted to a more positive potential, which was consistent with an open-channel blockade. Paralleling hKv1.3 inhibition, curcumin significantly inhibited proliferation and interferon-γ secretion of T(EM) cells. Our findings demonstrate that curcumin is able to inhibit proliferation and proinflammatory cytokine secretion of T(EM) cells probably through inhibition of hKv1.3 channels, which contributes to the potency of curcumin for the treatment of autoimmune diseases. This is probably one of pharmacological mechanisms of curcumin used to treat autoimmune diseases.

[Controlled study on different acupoint-prescription for the acupoint catgut embedding therapy in treatment of bronchial asthma].

OBJECTIVE: To probe into the best acupoint-prescription for the simple acupoint catgut embedding therapy in treatment of bronchial asthma. METHODS: Forty-eight Wistar rats were randomly divided into six groups: the simple acupoint catgut embedding at Feishu (BL 13), Danzhong (CV 17) and Shenshu (BL 23) group (group A), the simple acupoint catgut embedding at Shenshu (BL 23) group (group B), the simple acupoint catgut embedding at Feishu (BL 13) and Danzhong (CV 17) group (group C), Dexamethasone group (group D), model group (group E), and control group (group F), 8 rats in each group. The asthmatic models were established by Ovalbumin (OVA) except group F. Rats in group A, B and C were treated with catgut embedding at the corresponding acupoints from the first experimental day. In group E and D, Dexamethasone and sterile were intraperitoneal injected respectively from the 15th experimental day, once a day for 2 weeks consistantly. No interventions were added on group F. For the six groups, the symptoms of asthmatic attack were observed and the pathologic changes of lung tissue were examined. RESULTS: (1) The times of sneeze and rhinocnesmus scratching nose in group E were increased significantly compared with those in group F (both P < 0.01), but there was no significant difference between group D and F (both P > 0.05). As compared with that in group E, except the times of sneeze in group C, the times mentioned above were decreased significantly in all the treatment groups (all P < 0.01). There was no significant difference between group A and D (both P > 0.05), but the times mentioned above were increased significantly in group B and C as compared with that in group A (all P < 0.01). (2) The symptoms of asthmatic attack and the pathologic changes of airway tissue were all alleviated in group A, B and C, but a better amelioration was observed in group A, with no mucus epistom in the bronchial lumen and few infiltrations of inflammatory cells around the bronchi. CONCLUSION: The improvement of the simple acupoint catgut embedding at "Feishu" (BL 13), "Danzhong" (CV 17)and "Shenshu" (BL 23) on the airway inflammation in asthmatic rats is better than that of catgut embedding at "Feishu" (BL 13) and "Danzhong" (CV 17) or at "Shenshu" (BL 23) only.

[Study on changes in cytokines of infertile women with mycoplasma infection and intervention with traditional Chinese medicines].

OBJECTIVE: To investigate the changes in cytokines (IL-1beta, IL-2, TNF-alpha) of peripheral blood and cervical mucous of infertile women with mycoplasma infection and the effect of intervention of traditional Chinese medicines (TCMs). METHOD: According to the results of culture of mycoplasma from genital tracts, 72 patients with positive mycoplasma were randomly divided into the TCM group (38 cases) and the western medicine group (34 cases). The western medicine group was treated with 0.5 g azithromycin for 3 days and consecutively treated for six courses of treatment, each course of treatment of 4 days. The TCM group were treated with Xiaozhi decoction twice every day for 6 weeks. The IL-1beta, IL-2 and TNF-alpha levels of the peripheral blood and cervical mucous of the two groups were measured by the Ria testing before and after the treatment, and the mycoplasma culture (-) of 32 infertile women as set for control. RESULT: Before the treatment, TNF-alpha and IL-1beta in levels of the two treatment groups were higher than those of the control group (P < 0.01). In the TCM group, TNF-alpha and IL-1beta levels showed significant differences compared with those before the treatment (P < 0.05) and those of the western group after the treatment (P < 0.01); and IL-2 level didn't have significant change before and after the treatment. The cytokines in peripheral blood of the two treatment groups showed notable difference compared with those of the control group (P < 0.01). In TCM group, IL-2 level had remarkable difference compared with that before the treatment (P < 0.01) and compared with the control group after the treatment (P < 0.01). CONCLUSION: Cytokines (IL-1beta, IL-2, TNF-alpha) in the peripheral blood and cervical mucous increase in infertile women with the mycoplasma infection, suggesting that TCMs can effectively inhibit the levels of IL-1beta, IL-2, TNF-alpha in the peripheral blood and IL-1beta, TNF-alpha in cervical mucous. It is proved that Xiaozhi decoction can be used to treat infertile women with mycoplasma infection.

[Effect of needle-implantation at "Neiguan" (PC 6) on the expression of myocardial TGF-beta3 and its mRNA in the Chinese miniswine with myocardial ischemia injury].

OBJECTIVE: To observe the effect of needle-implantation (NI) at "Neiguan" (PC 6) on the expression of myocardial transforming growth factor-beta 3 Protein(TGF-beta 3) and mRNA in Chinese miniswine with myocardial ischemia (MI) injury. METHODS: A total of 32 Chinese Guizhou miniswine were randomly and equally divided into Sham-operation group, model group, NI-PC 6 group and NI-Geshu (BL17, NI-BL17) group. MI injury model was established by occlusion of the descending anterior branch of the left coronary artery. Two acupuncture needles (veterinary use) were separately and subcutaneously implanted into "Neiguan" (PC 6) and "Geshu" (BL17) areas for 7 days. TGF-beta 3 protein and mRNA expressions were determined by Western blot and real-time PCR techniques, separately. RESULTS: In comparison with the sham-operation (sham) group, TGF-beta 3 protein and mRNA expressions in model group were upregulated significantly (P < 0.05). While compared with the model group, myocardial TGF-beta3 mRNA expression was upregulated considerably in NI-PC 6 group (P < 0.01), rather than in NI-BL17 group (P > 0.05), and myocardial TGF-beta 3 expression in both NI-PC 6 and NI-BL17 groups was upregulated obviously (P < 0.01). Comparison between NI-PC 6 and NI-BL17 groups showed that the expression levels of myocardial TGF-beta 3 protein and mRNA were significantly higher in NI-PC 6 group than in NI-BL17 group (P < 0.05). CONCLUSION: Needle-implantation of "Neiguan" (PC 6) can upregulate myocardial TGF-beta 3 protein and mRNA expression in MI Chinese miniswine, which may contribute to its effect in improving the ischemic myocardial injury by way of enhancing the angiopoiesis.

[Influence of electroacupuncture on myocardial NO and NOS and intracellular Ca2+ contents in myocardial ischemia-reperfusion injury rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on contents of myocardial nitric oxide (NO), nitric oxide synthase (NOS), and intracellular Ca2+ levels in experimental myocardial ischemia-reperfusion injury (MI/RI) rats, so as to provide some evidence for acupuncture treatment of cardiovascular disorders and for the correlation between the acupoints and the internal organs. METHODS: Fifty male Wistar rats were randomly divided into sham-operation (sham), model, Neiguan (PC 6), Lieque (LU 7), and Hegu (LI 4) groups (n = 10/group). MI/RI model was established by occlusion of the descending anterior branch of the left coronary artery for 40 min and reperfusion for 60 min. EA (30 Hz/100 Hz, 2-4 mA) was applied to the bilateral PC 6, LU 7 and LI 4 for 20 min. The myocardial tissue of the ischemic area was taken, homogenated and centrifuged, separated for assaying NO and NOS activity by using spectrophotometer and their contents with nitrate reductase colorimetric assay. Another part of the myocardium tissue was processed to be cell suspension (added with Fluo-3/AM dye) for assaying intracellular free [Ca2+]i concentration under the laser confocal microscope. RESULTS: Compared with sham group, myocardial NO and NOS contents of model group decreased mildly and significantly (P < 0.05), while myocardial fluorescence intensity (content) of intracellular Ca2+ increased evidently (P < 0.01). In comparison with model group, the contents of myocardial NO and NOS were upregulated considerably in PC 6 group (P < 0.05), and intracellular Ca2+ content in PC 6 group reduced significantly (P < 0.01). No significant differences were found between LU 7 and model groups and between LI 4 and model groups in the above mentioned 3 indexes (P > 0.05). CONCLUSION: EA of PC 6 can upregulate myocardial NO and NOS contents and downregulate myocardial intracellular Ca2+ level in MI/RI rats, which may contribute to its effect in relieving myocardial injury. Neiguan (PC 6) and the heart have a relatively specific correlation.

[Effect of the simple acupoint catgut embedding on the expression of ICAM-1, NF-kappaB and airway inflammation in rats with asthma].

OBJECTIVE: To explore the underlying mechanism of the simple acupoint catgut emhedding on the treatment of the asthma in rats. METHODS: Thirty two Wistar rats with asthma were estahlished hy ovalhumin (OVA), and randomly divided into four groups: the simple acupoint catgut emhedding group (A), Dexamethasone group (B), model group (C), control group (D), 8 cases in each group. In group A, emhed thread was carried out on the first experimental day, "Feishu (BL 13)" "Danzhong (CV 17)" "Shenshu (BL 23)" were selected. In the group B and the group C, 1.5 mg/kg Dexamethasone and 20 mg/kg sterile water were intraperitoneal injected respectively from the 15th experimental day, once a day for 2 weeks consistently. There was no treatment in the group D. For the four groups, asthmatic attack was ohserved, the cells in hronchoalveolar lavage fluid were counted and classified, the pathologic change of lung tissue was examined, the level of ICAM-1 and NF-kappaB in hronchioles was detected with immunohistochemistry. RESULTS: (1) In the group A, the symptom of asthmatic attack was alleviated and the infiltration of inflammatory cells around the hronchi were remarkahly decreased. In the group C, the total amount of cells in hronchoalveolar lavage fluid and other inflammatory cells were ohviously higher than those of group A. On the other hand, inflammatory effusion in airway and infiltration of inflammatory cells in airway wall in the group A were lower than those of group C. (2) The expression of ICAM-1 and NP-kappaB in lung tissue of group C was higher than those of group D (P < 0.01). (3) The expression of ICAM-1 and NF-kappaB in lung tissue of group B and group A was ohviously lower than those of group C (P < 0.05, P < 0.01). CONCLUSION: The simple acupoint catgut emhedding can inhahit the expression of ICAM-1 and NP-kappaB and alleviate airway inflammation in the asthmatic rats.

Protective effect of Astragalus polysaccharides on ATP binding cassette transporter A1 in THP-1 derived foam cells exposed to tumor necrosis factor-alpha.

Astragalus polysaccharide (APS), the main extract from the traditional Chinese medicinal herb Astragalus membranaceus, has been reported to benefit the treatment of immune-inflammatory diseases and metabolic disorders. In atherosclerotic plaques, proinflammatory cytokines exert adverse effects on lipids thereby aggravating atherosclerosis. Recent evidence shows that tumor necrosis factor-alpha (TNF-alpha) can down-regulate the expression of ATP-binding cassette transporter A1 (ABCA1), which plays a vital role in reverse cholesterol transport and determines the process of atherosclerosis. In the present study, the effects of APS on ABCA1 expression, cholesterol effluent rate and total cholesterol content of THP-1 derived foam cells exposed to TNF-alpha were investigated. Compared with the foam cells exposed to TNF-alpha, ABCA1 expression was promoted in the presence of APS. Consequently the cholesterol effluent rate increased and the total cholesterol content decreased significantly. TNF-alpha could enhance the activity of nuclear factor-kappa B (NF-kappaB) in the foam cells. This effect could be attenuated by APS. These findings suggest that APS could protect ABCA1 against the lesion of TNF-alpha in THP-1 derived foam cells, which may contribute to its antiatherosclerotic properties.

[Effect of electroacupuncture of "Neiguan" (PC 6) on Na+-K+-ATPase activity and its gene expression in cardiocyte membrane in rats with myocardial ischemia-reperfusion injury].

OBJECTIVE: To compare the effects of electroacupuncture (EA) of "Neiguan" and other acupoints on Na+ -K+ -ATPase activity and its gene expression in ischemic myocrardial cell membrane in rats. METHODS: A total of 50 Wistar rats were randomly divided into pseudo-operation (control), model, Neiguan (PC6), Shenmen (HT7) and Hegu (LI4) groups, with 10 rats in each group. Myocardial ischemia-reperfusion (I/R) model was duplicated by occlusion of the descending anterior branch of the left coronary artery and reperfusion. EA (30/100 Hz, 2-4 mA) was applied to the above-mentioned acupoints for 20 min respectively. Na+ -K+ -ATPase activity was determined by inorganic phosphorus colorimetry, and Na+ -K+ -ATPase gene expression was detected using reverse transcription polymerase chain reaction (RT-PCR) technique. RESULTS: Compared with control group, the activity of Na+ -K+ -ATPase in model, PC6, HT7 and LI4 groups decreased significantly after myocardial I/R (P<0.05, 0.01); while in comparison with model, the activity of Na+ -K+ -ATPase in PC6 group increased considerably (P<0.01). Regarding the expression of Na+ -K+ -ATPase mRNA, compared with control group, it was down-regulated significantly in model, HT7 and LI4 groups (P<0.01); while in PC6 group, Na+ -K+ -ATPase mRNA expression was up-regulated markedly compared with model group (P<0.05); The activity of Na+ -K+ -ATPase and the expression of Na+ -K+ -ATPase mRNA in HT7 and LI4 groups were significantly lower than those in PC6 group (P<0.01, 0.05). No significant differences were found between HT7 and LI4 groups in the activity of Na+ -K+ -ATPase and the expression of Na+ -K+ -ATPase mRNA (P>0.05). CONCLUSION: EA of "Neiguan" (PC6) may potentiate the activity of Na+ -K+ -ATPase and up-regulate Na+ -K+ -ATPase mRNA expression, which may contribute to its protective effect on ischemic cardiocytes.

[Comparative study on effects of static pressure stimulation on release of PGE2 and IL-6 in fibroblasts in the rat "Zusanli" and its adjacent areas].

OBJECTIVE: To provide some possible theoretical and experimental basis for the modern biomedical mechanism of "receiving stimulation, preventing and treating diseases", by exploring the effects of pressure signal's biological transformation of rat "Zusanli" (ST 36) fascia tissue fibroblasts under mechanical stimulation. METHODS: Pressure was given on rat "Zusanli" (ST 36) and its adjacent area's fascia tissue cells cultured in vitro and identified by morphology. The contents of prostaglandin E2 (PGE2) and interleukin-6 (IL-6) in culture medium were detected. RESULTS: The fascia tissue cells in "Zusanli" and its adjacent area are almost consisted of fibroblasts. The pressure stimulation significantly accelerated the synthesis and release of PGE2 and IL-6. CONCLUSION: Acupoint and non-acupoint fibroblasts can directly receive mechanical stimulation, and then the mechanical signals were transformed as biological ones.

[Experimental study on action of acupoint embedding thread on ANCA in the rat of ulcerative colitis].

OBJECTIVE: To observe therapeutic effect of acupoint embedding thread therapy on ulcerative colitis and to study the mechanism. METHODS: The rat model of ulcerative colitis was developed with 2,4-dinitrochlorobenzene method. The 20th day after the end of model development, they were randomly divided into a model group, a salicylazosulfapyridine (SASP) treatment group and an acupoint embedding thread therapy group. The SASP treatment group were treated with intragastrical perfusion of 4.5% SASP suspension, and the acupoint embedding thread therapy group were treated with embedding thread at "Zusanli" (ST 36) and "Shangjuxu" (ST 37). Anti-neutrophil cytoplasmic antibody (ANCA) was detected with indirect immunofluorescence, and the pathological changes were investigated with pathological tissue section technique. RESULTS: The positive rate of ANCA reached to 62.5% in the rat with ulcerative colitis with significant differences among the 3 groups. After treatment, the positive rate of ANCA was decreased in the two treatment groups (P < 0.05, P < 0.01). CONCLUSION: Acupoint embedding thread therapy has definite therapeutic effect on ulcerative colitis and the mechanism is possibly related with the decrease of ANCA.