石菖蒲挥发油、β-细辛醚对高脂血症大鼠血管舒缩与抗血小板聚集的作用

目的研究石菖蒲挥发油、β-细辛醚对高脂血症模型动物血管舒缩与抗血小板聚集的作用。方法大鼠喂养高脂饲料造成高脂血症模型,采用流式细胞术检测血液中 CD_(62p)、CD_(63)表达率,并用放免法测定脑组织中内皮素(ET)、降钙素基因相关肽(CGRP)、神经肽 Y(NPY)的舍量。结果石菖蒲挥发油能降低脑ET 的含量而升高脑 CGRP 的浓度;β-细辛醚能降低血 CD_(62p)表达率、降低脑 ET、NPY 的含量。结论石菖蒲挥发油、β-细辛醚有舒张血管、抗血小板聚集活性的作用。

Investigation of Calcitonin and Calcitonin Gene-related Peptide in 88 Cases of Medullary Thyroid Carcinoma

Objective： To investigate the changes of calcitonin （CT） and calcitonin gene-related peptide （CGRP） in patients with medullary thyroid carcinoma （MTC）. Methods： Fifty-eight cases of MTC were selected and the relationship between the CT levels and metastasis was investigated. The immunohistochemical method was used to detect the expression of CT and CGRP in the 58 samples of MTC tissues. The CT and CGRP in 30 newly diagnosed MTC inpatients were measured before operation and in the first few days after operation using a radioimmunoassy. Results： （1） The rate of residual tumor had a significant difference between the normal serum CT group one month after operation and the elevated group at the same period （P〈0.01）. （2） Immunohistochemical study revealed the positive rate of CT was about 98%, and that of the CGRP was 87.8%. （3） Part of the patients had an elevated CGRP levels while CT levels was normal. （4） The serum CT levels were decreased to a stable range one week after operation. Conclusion： CT is a useful index to evaluate the efficacy of surgical treatment. The measurement of serum CGRP is helpful in the diagnosis of MTC, especially for those whose preoperative CT levels are normal.

Therapeutic effects of Clostridium butyricum on experimental colitis induced by oxazolone in rats

AIM： To evaluate the therapeutic effects of a probiotic supplement （Clostridium butyricum, CGMCC0313 ） in a chemically-induced rat model of experimental colitis. METHODS： An experimental ulcerative colitis model was established by rectal injection of oxazolone into the colon of 40 Wistar rats randomly divided into four groups. The positive control group was sacrificed 3 d after colitis onset. The remaining groups were fed daily with either 2 mL of C. butyricum （2.3 × 10^11 CFU/L）, 2 mL of mesalamine （100 g/L）, or 1 mL of sodium butyrate （50 mmol/L） for 21 d. The animals＇ body weight, behavior, and bowel movements were recorded weekly. After sacrifice, visual and microscopic observations of pathological changes of colon tissue were made, body weight and wet colon mass index were measured and recorded, and serum levels of interleukin-23 （IL-23） and TNF-α were measured using ELISA. Expression of calcitonin gene-related peptide in colon tissue was measured by RT-PCR. Finally, changes in rat intestinal microflora status were measured in all groups.lowered the serum levels of both IL-23 and tumor necrosis factor-α （TNF-α） with similar or even better efficiency than that of mesalamine or sodium butyrate. The rat intestinal flora appeared to recover more quickly in the group treated with C. butyricum than in the mesalamine and sodium butyrate groups. Finally, we found that the expression level of calcitonin gene related peptide was elevated in colon tissue in the sodium butyrate treated group but not in the C. butyricum or mesalamine treated groups, indicating a sensitization of colon following sodium butyrate treatment. CONCLUSION： In our experimental colitis model, treatment with C. butyricum CGMCC0313, a probiotic supplement, is at least as efficient as treatment with mesalamine.

Protection of Calcitonin Gene-Related Peptide-Induced Preconditioning against Myocardial Injury due to Adriamycin in the Isolated Rat Heart *

The protective effect of calcitonin gene-related peptide (CGRP)-induced preconditioning on myocardial injury due to adriamycin was studied in the isolated perfused rat heart. Adriamycin (100 and 200 (mol / L) caused a gradual decrease in coronary flow (CF) and cardiac function (LVP and LV dp/dtmax), and an increase in the level of MDA. Pretreatment with CGRP at the concentration of 5 nmol/L for 5 min markedly reduced the attenuation of CF and cardiac function and inhibited the elevation of MDA content induced by adriamycin. The findings suggest that the pretreatment with CGRP possesses a protection against myocardial injury elicited by adriamycin. The present results also suggest that the protection of CGRP may be related to a reduction in lipid peroxidation.

Nitric oxide-releasing aspirin but not conventional aspirin improves healing of experimental colitis

AIM:To determine the effect of non-selective cyclooxygenase (COX) inhibitors,selective COX-2 inhibitors and nitric oxide (NO)-releasing aspirin in the healing of ulcerative colitis.METHODS:Rats with 2,4,6 trinitrobenzenesulfonic acid (TNBS)-induced colitis received intragastric (ig) treatment with vehicle,aspirin (ASA) (a nonselective COX inhibitor),celecoxib (a selective COX-2 inhibitor) or NO-releasing ASA for a period of ten days.The area of colonic lesions,colonic blood flow (CBF),myeloperoxidase (MPO) activity and expression of proinflammatory markers COX-2,inducible form of nitric oxide synthase (iNOS),IL-1β and tumor necrosis factor (TNF)-α were assessed.The effects of glyceryl trinitrate (GTN),a NO donor,and 2-(4-carboxyphenyl)-4,5-dihydro-4,4,5,5-tetramethyl-1H-imidazolyl-1-oxy-3-oxide,onopotassium salt (carboxy-PTIO),a NO scavenger,administered without and with ASA or NO-ASA,and the involvement of capsaicin-sensitive afferent nerves in the mechanism of healing the experimental colitis was also determined.RESULTS:Rats with colitis developed macroscopic and microscopic colonic lesions accompanied by a significant decrease in the CBF,a significant rise in colonic weight,MPO activity and plasma IL-1β and TNF-α levels.These effects were aggravated by ASA and 5-(4-chlorophenyl)-1-(4-methoxyphenyl)3-(trifluoromethyl)-1H-pyrazole (SC-560),but not celecoxib and counteracted by concurrent treatment with a synthetic prostaglandin E 2 (PGE 2) analog.Treatment with NO-ASA dose-dependently accelerated colonic healing followed by a rise in plasma NO x content and CBF,suppression of MPO and downregulation of COX-2,iNOS,IL-1β and TNF-α mRNAs.Treatment with GTN,the NO donor,significantly inhibited the ASA-induced colonic lesions and increased CBF,while carboxy-PTIO or capsaicin-denervation counteracted the NO-ASAinduced improvement of colonic healing and the accompanying increase in the CBF.These effects were restored by co-treatment with calcitonin gene related peptide (CGRP) and NO-ASA in capsaicin-denervated animals.CONCLUSION:NO-releasing ASA,in contrast to ASA,COX-1 inhibitors,and SC-560,accelerated the healing of colitis via a mechanism involving NO mediated improvement of microcirculation and activation of sensory nerves releasing CGRP.

EFFECT OF ACUPUNCTURE OF "ZUSANLI" AND "WEISHU" ACUPOINTS ON EXPERIMENTAL GASTRIC ULCER

Objective: To observe the therapeutic effect of electroacupuncture (EA) of "Zusanli"(ST 36) and "Weishu"(BL 21)on the experimental gastric ulcer in Xu period of a day for analyzing the best opportunity of acupuncture treatment of gastric ulcer. Methods:49 Wistar rats were randomly divided into ① control group, ② ST 36 Xu group, ③ BL 21 Xu group, ④ ST 36 Chen group, ⑤ BL 21 Chen group, ⑥ ST 36 IT group, and ⑦ BL 21 IT group, with 7 rats in each group. Rat gastric ulcer model was established using intra gastric administration of glacial acetic acid. Gastric mucosal thickness,mucosal muscular defect width,superficial mucosal mucus index (MI),intramucosal MI ,Total MI,plasma 6 keto PGF1α ,serum NO and gastric tissue CGRP concentrations were used as the indexes. The EA treatment was conducted once daily at Xu period (7-9 o’clock in the evening), Chen period (8-10 o’clock in the morning) and irregular time (IT) with 6 days being a therapeutic course, two courses altogether. Results:After EA treatment,①the regenerated mucosal thickness values of the aforementioned 6 EA groups, particularly ST 36 Xu group and BL 21 Xu group, were all apparently higher than that of control group (P<0.05-0.01), while the mucosal muscle layer defect width values of different EA groups, particularly ST 36 Xu group and BL 21 Xu group, were strikingly lower than that of control group (P<0.05-0.01); ②the 3 MI of the 6 EA groups, particularly those of ST 36 Xu group were all significantly higher than those of control group (P<0.05-0.01); ③plasma 6 keto PGF1α contents of the 6 EA groups, particularly those of ST 36 Xu and BL 21 Xu groups, were all significantly higher than that of control group (P<0.05-0.01); ④serum NO and gastric tissue CGRP concentrations of the 6 EA groups, particularly those of ST 36 Xu and BL 21 Xu groups and CGRP concentration of ST 36 Chen group, were all significantly higher than those of control group (P<0.05-0.01);and ⑤there were positive correlation between superficial MI or total MI and NO level (r=0.858, 0.987, P<0.05,and 0.01),and between the superficial mucus index and CGRP concentration (r=0.9051, t=4.68, P<0.01), suggesting that the increase of the secretion function of the regenerated mucosal mucus is related to acupuncture induced increase of release of both NO and CGRP.Conclusion:The therapeutic effect of EA of ST 36 and BL 21 in Xu period is better in the treatment of gastric ulcer,and that of EA of ST 36 is the best.

Regulatory effects of insulin-like growth factor-1 on the expression of sensory neuropeptide mRNAs in cultured dorsal root ganglion neurons with excitotoxicity induced by glutamate

Objective To determine the effects of insulin-like growth factor-1 （IGF-1） on the expression of preprotachykinin （PPT） mRNA encoding substance P （SP） and calcitonin gene-related peptide （CGRP） mRNA in cultured dorsal root ganglion （DRG） neurons with excitotoxicity induced by glutamate （Glu）. Methods DRGs were dissected from embryonic day 15 Wistar rats. DRG neurons were dissociated and cultured for 48 h and then exposed to Glu （0.2 mmol/L） or Glu （0.2 mmol/L） plus IGF- 1 （5 nmol/L, 10 nmol/L and 20 nmol/L） for 12 h. The DRG neurons in control group were exposed to only growth media throughout the experiment. After that, the living DRG neurons were observed under inverted phase contrast microscope and microphotographs were taken. The expression levels of PPT and CGRP mRNAs were detected by reverse transcriptionpolymerase chain reaction （RT-PCR）. Results IGF-1 could inhibit Glu-induced shortening of neurite. Besides, IGF-1 could significantly increase the levels ofPPT mRNA and CGRP mRNA in primary cultured DRG neurons with Glu-induced excitotoxicity, in a dose-dependent manner. Conclusion IGF-1 may exert neuroprotective effects on DRG neurons against Glu-induced excitotoxicity, probably through regulating the expression levels of PPT and CGRP mRNAs.

Effects of electroacupuncture at 2 and 100 Hz on rat type 2 diabetic neuropathic pain and hyperalgesia-related protein expression in the dorsal root ganglion

Objective： To investigate the analgesic effects of electroacupuncture （EA） at 2 and 100 Hz on type 2 diabetic neuropathic pain （DNP） and on the expressions of the P2X3 receptor and calcitonin gene-related peptide （CGRP） in the dorsal root ganglion （DRG）. Methods： Rat type 2 DNP was induced by a high calorie and high sugar diet fed for 7 weeks, plus a single intraperitoneal injection of streptozotocin （STZ） after 5 weeks. EA at 2 and 100 Hz was carried out once every day after 7 weeks for 7 consecutive days. Body weight, serum fasting insulin （FINS）, fasting blood glucose （FBG）, insulin sensitivity index （ISI）, and paw withdrawal latency （PWL） were measured. The expressions of L4-L6 DRG P2X3 receptors and CGRP were assessed by immunofluorescence. Results： Type 2 DNP was successfully induced as shown by the increased body weight, FINS, and FBG, as well as the reduced ISI and PWL. Expressions of P2X3 receptors and CGRP in L4-L6 DRGs increased. EA at both 2 and 100 Hz relieved type 2 DNP, but the analgesic effect of EA was stronger at 2 Hz. P2X3 receptor expression decreased in L4-L6 DRGs following EA at 2 Hz and in L5 and L6 DRGs following EA at 100 Hz. EA at both 2 and 100 Hz down-regulated CGRP overexpression in L4-L6 DRGs. Conclusions： These findings indicate that EA at 2 Hz is a good option for the management of type 2 DNP. The EA effect may be related to its down-regulation of the overexpressions of the DRG P2X3 receptors and CGRP in this condition.

Intermedin/adrenomedullin2: an autocrine/paracrine factor in vascular homeostasis and disease

Intermedin(IMD)or adrenomedullin 2 is a novel peptide related to the calcitonin gene-related peptide(CGRP)family.Via calcitonin receptor-like receptor/receptor activity modifying proteins,the common receptor complexes of CGRP,IMD exerts a wide range of biological effects,especially regulation of cardiovascular homeostasis.Proteolytic processing of a larger IMD precursor yields a series of biologically active C-terminal fragments,IMD1–53,IMD1–47 and IMD8–47.IMD and its receptors are present in the cardiovascular system,and IMD is present at low levels in plasma.In the cardiovascular system,IMD has multiple functions such as regulation of blood pressure and cardiac function,pro-angiogenesis,endothelial barrier function protection,anti-oxidative stress,and anti-endoplasmic reticulum stress.IMD participates widely in the pathogenesis of atherosclerosis,hypertension,pulmonary arterial hypertension and vascular calcification.It is a vascular regulatory factor of homeostasis and a vital endogenous protective factor against vascular diseases.

Effect of Qilongtoutong granule on calcitonin gene-related peptide,beta-endorphin, serotonin, dopamine, and noradrenalin in migraine model rats and mice

OBJECTIVE： To study the effect of Qilongtoutong granule （QLTT） on plasma calcitonin gene-related peptide （CGRP）, beta-endorphin （I[3-EP）, 5-HT, dopa- mine （DA）, noradrenalin （NE）, and blood viscosity in migraine model rats and mice. METHODS： Both the acute blood stasis model group and nitroglycerin-induced migraine model group included 60 Sprague-Dawley rats. The reser- pine-reduced model group had 60 Kunming mice. Rats from each test were grouped into normal con- trol group, model group, Zhengtian pill （ZTP） group, and high, moderate, or low-dose QLTT groups. In the acute blood stasis model test, after gavage for 7 days, rats were given 0.8 mL/kg adren- aline hydrochloride subcutaneously twice, and kept in ice water for 5 min. After fasting for 12 h, rats were anesthetized and blood samples were collected for detection of blood viscosity. In the nitro- glycerin-induced migraine group, after gavage for 7 days, rats were intraperitoneally injected nitro- glycerin （10 mg/kg）, and 4 h later, blood samples were collected from postcava for measuring the plasma CGRP and 13-EP levels. In the reserpine-re- duced model test, except the normal control group, mice were administered reserpine （0.25 mg/ kg, i.h.） for 9 days. Mice received intragastric admin- istration from the third day to the ninth day. One hour after the last gavage, blood and brain tissue samples were obtained. Then, blood clotting time and the contents of neurotransmitters were deter- mined. RESULTS： QLTT- （3.6, 1.8, and 0.9 g/kg） and ZTP-treated rats had lower blood viscosity than that in model rats under different shear rates （P〈 0.01）. QLTT- （3.6, 1.8 g/kg） and ZTP-treated rats had significantly lower plasma CGRP levels and higher plasma 13-EP levels than those in model rats （P〈 0.01）. QLTT treatment at dose of 0.9 g/kg had lower plasma CGRP levels as well （P〈0.05）. QLTT- （5.2, 2.6 g/kg） and ZTP-treated mice had longer blood clotting time than that in model mice （P〈0.01）. QLTT- （2.6 g/kg） and ZTP-treated mice had higher plasma serotonin （5-HT） levels than those in model mice （P〈0.05）. CONCLUSION： QLTT-treated animals had lower plasma CGRP level, higher plasma 13-EP, 5-HT, high- er brain tissue 5-HT, NE, DA levels, and lower blood viscosity than those in the migraine model animals.

Electroacupuncture for functional dyspepsia and the influence on serum Ghrelin, CGRP and GLP-1 levels

Objective： To observe the clinical efficacy of electroacupuncture for functional dyspepsia（FD）, and explore the corresponding mechanism.Methods： Sixty-four FD patients were randomly divided into electroacupuncture group and western medicine group, with 32 cases in each group. In electroacupuncture group, electroacupuncture at Zusanli（足三里ST 36）,Sanyinjiao（三阴交SP 6）,Gongsun（公孙SP 4） and Neiguan（内关PC 6） was performed for once a day, and the needles were retained for 30 min. In western medicine group, oral administration of mosapride citrate dispersible tablets in a dosage of 5 mg/time was carried out for 3 times a day. Treatment was conducted for 30 consecutive days in both groups. The scores of Leeds dyspepsia questionnaire（LDQ） and functional digestive disorder quality of life（FDDQL） of patients in both groups were recorded before and after treatment. Serum Ghrelin, CGRP and GLP-1 levels of patients were tested before and after treatment respectively, and the clinical efficacy of patients in both groups was evaluated after treatment.Results： In western medicine group, LDQ score after treatment was lower than that before treatment（P 0.05）, FDDQL score after treatment was higher than that before treatment, while the differences were not statistically significant（P0.05）. LDQ score in electroacupuncture group after treatment was lower than that before treatment（P0.05）, and also lower than that in western medicine group at the same time point（P 0.05）. FDDQL score in electroacupuncture group after treatment was higher than that before treatment（P0.05）, and also higher than that in western medicine group at the same time point（P0.05）. In western medicine group, Ghrelin level after treatment was higher than that before treatment（P 0.05）, CGRP level reduced, and the differences were not statistically significant（P 0.05）. GLP-1 level after treatment was also higher than that before treatment（P0.05）. In electroacupuncture group,Ghrelin level after treatment was higher than that before treatment, CGRP level reduced, and GLP-1 level after treatment was also higher than that before treatment（both P 0.05）. According to the comparison of values of each index between electroacupuncture group and western medicine group after treatment,the differences were all statistically significant（all P 0.05）. The total effective rate in electroacupuncture group was 90.63%（29/32） which was higher than that in western medicine group 68.75%（22/32）, and the differences were statistically significant（P0.05）.Conclusion： Electroacupuncture at ST 36, SP 6, SP 4 and PC 6 can effectively improve the clinical symptoms of FD patients, and the mechanism might be related with the increase of serum Ghrelin and GLP-1 levels and the decrease of serum CGRP level.

Modification at Lys24 with fatty acid generates active calcitonin gene-related peptide with much improved plasma stability

As a 37-amino acid vasoactive neuropeptide, calcitonin gene-related peptide （CGRP） is widely distributed in nervous systems. The studies and clinical applications of CGRP are limited by its peptide nature and short half-life. A series of peptide analogues of the α-form of CGRP were synthesized. Afterwards, by using in vitro metabolic and activity studies, we prepared two high affinity analogues with significantly improved plasma stability.