The role of NO-cGMP pathway inhibition in vascular endothelial-dependent smooth muscle relaxation disorder of AT1-AA positive rats: protective effects of adiponectin.

Angiotensin II type 1 receptor autoantibodies (AT1-AA) cause endothelial-dependent smooth muscle relaxation disorder. It is well understood that impairment of the NO-cGMP signaling pathway is one of the mechanisms of endothelial-dependent smooth muscle relaxation disorder. However, it is still unclear whether AT1-AA induces endothelial-dependent smooth muscle relaxation disorder via the impairment of the NO-cGMP signaling pathway. In addition, adiponectin exerts vascular endothelial protection through the NO-cGMP signaling pathway. Therefore, the purpose of this investigation was to assess the mechanism of vascular endothelial-dependent smooth muscle relaxation disorder induced by AT1-AA and the role of adiponectin in attenuating this dysregulation. Serum endothelin-1 (ET-1), adiponectin and AT1-AA were detected by enzyme-linked immunosorbent assay. In preeclamptic patients, there was an increased level of AT1-AA, which was positively correlated with ET-1 and negatively correlated with adiponectin, as elevated levels of ET-1 suggested endothelial injury. AT1-AA-positive animal models were actively immunized with the second extracellular loop of the angiotensin II type 1 receptor (AT1R-ECII) for eight weeks. In thoracic aortas of AT1-AA positive rats, ET-1 was elevated, endothelium-dependent vasodilation was decreased. Paradoxically, as the upstream element of the NO-cGMP signaling pathway, NO production was not decreased but increased, and the ratio of p-VASP/VASP, an established biochemical endpoint of NO-cGMP signaling pathway, was reduced. Moreover, the levels of nitrotyrosine and gp91phox which indicate the presence of peroxynitrite (ONOO-) and superoxide anion (O2·-) were increased. Pretreatment with the ONOO- scavenger FeTMPyP or O2·-scavenger Tempol normalized vasorelaxation. Key enzymes responsible for NO synthesis were also assessed. iNOS protein expression was increased, but p-eNOS(Ser1177)/eNOS was reduced. Preincubation with the iNOS inhibitor 1400 W or eNOS agonist nebivolol restored vasorelaxation. Further experiments showed levels of p-AMPKα (Thr172)/AMPKα, which controls iNOS expression and eNOS activity, to have been reduced. Furthermore, levels of the upstream component of AMPK, adiponectin, was reduced in sera of AT1-AA positive rats and supplementation of adiponectin significantly decreased ET-1 contents, improved endothelial-dependent vasodilation, reduced NO production, elevated p-VASP/VASP, inhibited protein expression of nitrotyrosine and gp91phox, reduced iNOS overexpression, and increased eNOS phosphorylation at Ser1177 in the thoracic aorta of AT1-AA positive rats. These results established that impairment NO-cGMP pathway may aggravate the endothelial-dependent smooth muscle relaxation disorder in AT1-AA positive rats and adiponectin improved endothelial-dependent smooth muscle relaxation disorder by enhancing NO-cGMP pathway. This discovery may shed a novel light on clinical treatment of vascular diseases associated with AT1-AA.

Adiponectin-mediated promotion of CD44 suppresses diabetic vascular inflammatory effects.

While adiponectin (APN) was known to significantly abolish the diabetic endothelial inflammatory response, the specific mechanisms have yet to be elucidated. Aortic vascular tissues from mice fed normal and high-fat diets (HFD) were analyzed by transcriptome analysis. GO functional annotation showed that APN inhibited vascular endothelial inflammation in an APPL1-dependent manner. We confirmed that activation of the Wnt/ß-catenin signaling plays a key role in APN-mediated anti-inflammation. Mechanistically, APN promoted APPL1/reptin complex formation and ß-catenin nuclear translocation. Simultaneously, we identified APN promoted the expression of CD44 by activating TCF/LEF in an APPL1-mediated manner. Clinically, the serum levels of APN and CD44 were decreased in diabetes; the levels of these two proteins were positively correlated. Functionally, treatment with CD44 C-terminal polypeptides protected diabetes-induced vascular endothelial inflammation in vivo. Collectively, we provided a roadmap for APN-inhibited vascular inflammatory effects and CD44 might represent potential targets against the diabetic endothelial inflammatory effect.

[Effect of acupuncture-moxibustion on negative emotions and plasma tryptophan metabolism in patients with Crohn's disease at active stage].

OBJECTIVE: To observe the effect of acupuncture-moxibustion on negative emotions and plasma tryptophan (Trip)-kynurenine (Kyn) metabolism in the patients with Crohn's disease (CD) at the mild and moderate active stage. METHODS: A total of 66 CD patients were randomized into an observation group (33 cases, 1 case dropped off) and a control group (33 cases, 2 cases dropped off). In the observation group, acupuncture was applied in combination with moxibustion. In the control group, the sham-acupuncture was used in combination with sham-moxibustion. In both of the observation group and the control group, acupuncture was applied to Zhongwan (CV 12), Shangjuxu (ST 37), Sanyinjiao (SP 6), Gongsun (SP 4), Hegu (LI 4), Quchi (LI 11), Taixi (KI 3) and Taichong (LR 3), and moxibustion was applied to Tianshu (ST 25) and Zusanli (ST 36). The treatment was given once every two days, 3 times a week, totally for 12 weeks. Separately, before and after treatment, the score of the hospital anxiety-depression scale (HADS) and the score of intestinal core symptoms (degree of abdominal pain and frequency of diarrhea) were observed in the patients of the two groups. The concentration of plasma indoleamine 2,3-dioxygenase 1 (IDO1) and the ratios of Kyn/Trp, QuinA/Kyn, KynA/Kyn and KynA/QuinA were compared between the two groups. RESULTS: Compared with before treatment, the scores of HADS-A and HADS-D in the observation group and the score of HADS-A in the control group were all reduced after treatment (P<0.01, P<0.05). The scores of abdominal pain degree in the two groups and score of diarrhea frequency in the observation group were all reduced after treatment (P<0.001). After treatment, the reducing ranges of the score of HADS-A and the scores of abdominal pain degree and diarrhea frequency in the observation group were all larger than the control group (P<0.01, P<0.05). Compared with before treatment, the plasma IDO1 concentration in the two groups and the ratios of plasma Kyn/Trp and QuinA/Kyn in the observation group were all reduced after treatment (P<0.001, P<0.05, P<0.01), the ratios of plasma KynA/Kyn and KynA/QuinA were increased after treatment in the observation group (P<0.05, P<0.01). After treatment, the changes in IDO1 concentration and the ratios of plasma QuinA/Kyn and KynA/QuinA in the observation were larger than the control group (P≤0.01, P<0.05). In the observation group, the difference in the ratio of plasma KynA/Kyn before and after treatment was negatively related to the improvement value of HADS-D (r =-0.67, P<0.05). After treatment, plasma IDO1 concentration was positively related to HADS-A in the observation group (r =0.65, P<0.05). CONCLUSION: Acupuncture and moxibustion relieve the negative emotions of anxiety and depression in CD patients at mild and moderate active stage, which is probably related to the regulation of plasma Trp-Kyn metabolic pathway.

[Efficacy of electroacupuncture nerve stimulation therapy for interstitial cystitis/bladder pain syndrome].

OBJECTIVE: To explore the clinical efficacy of electroacupuncture nerve stimulation therapy (ENST) for interstitial cystitis/painful bladder syndrome (IC/PBS). METHODS: A total of 68 patients with IC/PBS were randomly divided into an observation group and a control group, 34 cases in each one. The patients in the observation group were treated with ENST; abdominal four acupoints and sacral four acupoints were connected with a pair of electrodes and treated alternately every other day. The ENST was given 50 min per times, three times a week for 3 months. The patients in the control group were treated with perfusion therapy of four-medication combination (heparin sodinm, lidocaine, sodium bicarbonate, gentamicin sulfate), twice a week for the first 6-8 weeks, followed by twice per month for 3 months. The infusion fluid remained for 1 h before discharging. The O' Leary-Sant score, including interstitial cystitis symptom index (ICSI) and interstitial cystitis problem index (ICPI), 24 h urination frequency, visual analogue scale (VAS) and maximum bladder volume were observed before treatment and treatment of 1 month, 3 months and 6 months after treatment respectively; the adverse events during the treatment were also recorded. RESULTS: Compared before treatment, the O'Leary-Sant score (ICSI, ICPI), 24 h urination frequency, VAS and maximum bladder volume in the two groups were improved after 1, 3 months treatment and 6 months after treatment (all P<0.05). The scores of ICSI, ICPI, VAS and 24 h urination frequency in the observation group were significantly lower than those in the control group (P<0.05). The maximum bladder volume in the observation group was significantly higher than that in the control group (P<0.05). Six months after treatment, the total effective rate in the observation group was 87.5% (28/32), which was higher than 69.7% (23/33) in the control group (P<0.01). No significant adverse events occurred during the treatment. CONCLUSION: ENST could effectively relieve the clinical symptoms of IC/PBS, but its long-term efficacy needs further observation.

[Discussion on the direction of moxibustion].

The types of moxibustion are various and its therapeutic effect could be influenced by many factors. The clinical manipulation of moxibustion usually involves sequential order, indicating the significance of direction in moxibustion. From the relationship between moxibustion direction and propagated sensation along channel, moxibustion direction and directional supplementation and reducing and others, this article proposes moxibustion direction could be one of the key factors to influence the clinical efficacy of moxibustion. It is hoped to improve moxibustion and increase clinical efficacy.

[Active Crohn's disease treated with acupuncture and moxibustion:a randomized controlled trial].

OBJECTIVE: To observe the effects and safety of clinical common symptoms of active Crohn's disease at the mild and moderate stages treated with acupuncture and moxibustion. METHODS: One hundred and two patients were randomly divided into an observation group and a control group, 51 cases in each one. Herb-partition moxibustion and acupuncture were used in the observation group. Herbal cakes with Huanglian (coptis chinensis), Fuzi (radix aconiti carmichaeli), Rougui (cinnamomum cassia), Muxiang (radices saussureae), Honghua (carthamus tinctorious), Danshen (the root of red-rooted salvia) and Danggui (angelica sinensis) as the main ingredients were used on Tianshu (ST 25), Qihai (CV 6) and Zhongwan (CV 12); acupuncture was used at Zusanli (ST 36), Shangjuxu (ST 37), Sanyinjiao (SP 6), Taixi (KI 3), Gongsun (SP 4) and Taichong (LR 3). Bran-partition moxibustion and shallow acupuncture were applied in the control group. Wheat bran-partition moxibustion in the shape of cake was implemented on the same acupoints as the observation group; shallow acupuncture was used 1~2 cm next to the acupoints. Treatment was given three times a week for 12 weeks, 36 times totally. Clinical effects of eight symptoms were evaluated by TCM symptom score, and the symptoms included abdominal pain (degree, frequency, time), diarrhoea (number, shape and frequency), fatigue, anorexia, borborygmus andflatus, fear of cold, soreness and weakness of waist and knees, tenesmus. RESULTS: After 12-week treatment, the eight symptoms were improved apparently (all P<0.05), with the scores of abdominal pain (degree, frequency, time), diarrhoea (shape and frequency), fatigue and anorexia in the observation group decreasing more obviously (all P<0.01). There was no statistical significance about the other symptom scores between the two groups (all P>0.05). CONCLUSIONS: Herb-partition moxibustion combined with acupuncture achieve safety and efficacy for mild and moderate Crohn's disease, and can improve the clinical common symptoms. Furthermore, their effects on abdominal pain (degree, frequency, time), diarrhoea (shape and frequency), fatigue and anorexia are better than those of bran-partition moxibustion and shallow acupuncture.

[Advances and thinking about prevention and treatment of inflammatory bowel diseases by acupuncture and moxibustion].

OBJECTIVE: To provide scientific foundations for clinical and acupoints researches on acupuncture for treatment of inflammatory bowel diseases. METHODS: Review the correlated documents of clinical experimental researches issued from 1995 to 2005, and the clinical and experimental documents about remedy of inflammatory bowel diseases by acupuncture and moxibustion were summarized and analyzed. RESULTS: Acupuncture and moxibustion had significant therapeutic effects on inflammatory bowel diseases, and the studies on the mechanisms have achieved some progresses, but the designs and the methods of these studies need to be improved. CONCLUSION: Afterwards, the specificity of acupoints, and factors of influencing the specificity should be studied via effective diseases treated with acupuncture and moxibustion, so as sum up common regularity, rich and develop the theory about specificity of acupoints to guide clinical treatment of acupuncture and moxibustion.