ABSTRACT
Objective: To investigate the mechanisms of electroacupuncture (EA) at Zusanli (ST 36), Liangmen (ST 21) and Sanyinjiao (SP 6) in intervening diabetic gastroparesis (DGP) based on calcium-activated chloride channel. Methods: Forty Sprague-Dawley rats were randomly divided into four groups, including a normal control group (group A), a model group (group B), an EA group (group C) and a metoclopramide group (group D), with 10 rats in each group. A single intraperitoneal injection of 2％ streptozotocin (STZ) combined with 8-week high-glucose high-fat diet was used to establish a DGP rat model. After intervention, gastrointestinal propulsive rate was observed; the expression level of transmembrane protein 16A (TMEM16A) was examined by immunohistochemistry; the Ca2+ concentration in interstitial cells of Cajal (ICCs) was detected by immunofluorescence; and whole-cell patch-clamp technique was applied to detect the current intensity of calcium-activated chloride channel (ICaCC) in ICCs in gastric antrum. Results: After modeling, the blood glucose levels in group B, group C and group D were significantly increased compared with group A (all P<0.01); after intervention, compared with group B, the blood glucose levels in group C and group D were significantly decreased (P<0.05, P<0.01); the intra-group comparison of blood glucose level between after modeling and after intervention found significant difference only in group C (P<0.01). The gastrointestinal propulsive rates in group B, group C and group D were significantly different from that in group A (P<0.01 or P<0.05); the gastrointestinal propulsive rates were markedly higher in group C and group D than in group B (P<0.01, P<0.01). The expressions of TMEM16A in group B and group C were decreased compared with group A (P<0.01, P<0.05); the expressions of TMEM16A in group C and group D were increased compared with group B (P<0.01, P<0.05). The fluorescence intensity of Ca2+ was significantly lower in group B than in group A (P<0.01); the fluorescence intensity of Ca2+ was significantly higher in group C and group D than in group B (P<0.01, P<0.05). ICaCC in ICCs in group B was significantly decreased compared with group A; ICaCC in group C and group D were increased compared with group B. Conclusion: EA at Zusanli (ST 36), Liangmen (ST 21) and Sanyinjiao (SP 6) can significantly improve gastrointestinal motility in DGP rats by up-regulating the ICaCC in ICCs.
ABSTRACT
Objective Carvedilol (Cvd) has a potential cardioprotective effect against myocardial ischemia/reperfusion (I/R) injury but its molecular mechanism is not yet clarified. The present study aimed to investigate whether the mechanism of Cvd against myocardial I/R injury-induced cardiomyocyte apoptosis is associated with its protection of the myocardium by modulating the unfolded protein response (UPR).Methods Forty male SD rats were randomly divided into five groups of equal number, sham operation, I/R, I/R + UPR agonist dithiothreitol (I/R+DTT), I/R + DTT with Cvd pretreatment at 5 mg/kg (I/R+DTT+Cvd), and I/R with Cvd pretreatment at 5 mg/kg (I/R+Cvd). The myocardial infarct size (infarct area / area-at-risk, IA/AAR) was measured by TCC & Evans blue double staining, the left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) detected by echocardiography, the apoptosis of the myocardiocytes determined by TUNEL staining, the activation of the UPR signaling pathway and the expressions of Caspase-12 and Casoase-3 detected by Western blot, and the concentrations of LDH and CK-MB assayed with the detection kit, followed by a comprehensive evaluation of the effects of Cvd on myocardial I/R injury.Results Myocardial IA/AAR was significantly increased in the I/R+DTT group as compared with the sham operation control (\[54.1±3.28\] % vs \[24.25±3.19\] %, P<0.05), higher in the I/R+DTT and I/R+DTT+Cvd (P<0.05) but lower in the I/R+Cvd than in the I/R group (P<0.05), and lower in the I/R+DTT+Cvd than in the I/R+DTT group (P<0.05). In comparison with the sham operation control, all the other four groups showed significantly decreased LVEF and LVFS (P<0.05), both remarkably lower in the I/R+DTT (\[44.5±1.56\] % and \[19.2±2.23\] %) than in the I/R group (\[61.5±2.63\] % and \[28.4±1.42\] %) (P<0.05), but higher in the I/R+DTT+Cvd and I/R+Cvd groups (P<0.05). The apoptosis rate of the cardiomyocytes was markedly increased in the I/R, I/R+DTT, I/R+DTT+Cvd, and I/R+Cvd groups as compared with that in the sham operation control (\[24.4±2.65\]%, \[48.3±1.62\]%, \[32.6±1.28\] % and \[13.2±2.21\]% vs \[6.2±1.27\]%, P<0.05), higher in the I/R+DTT and I/R+DTT+Cvd (P<0.05) but lower in the I/R+Cvd than in the I/R group (P<0.05). Compared with the sham operation control, the other four groups exhibited significantly elevated levels of expressions of the GRP78, CHOP and ATF6 proteins (P<0.05), all markedly higher in the I/R+DTT (P<0.05) but lower in the I/R+Cvd (P<0.05) and that of GRP78 lower in the I/R+DTT+Cvd than in in the I/R group (P<0.05), and all lower in the I/R+DTT+Cvd than in the I/R+DTT group (P<0.05).Conclusion Carvedilol can significantly alleviate the apoptosis of cardiomyocytes, and its molecular mechanism is related to its inhibitory effect on the UPR pathway.
ABSTRACT
OBJECTIVE: To observe the effects of electroacupuncture (EA) on gastrointestinal motility and the ultrastructure of interstitial cells of Cajal (ICC) and the expressions of c-kit receptor protein and stem cell factor (SCF) mRNA in diabetic gastroparesis (DGP) rats, so as to explore its mechanism. METHODS: Fifty SD rats were randomly divided into normal, model, acupoint, non-acupoint and metoclopramide groups (n＝10 rats/group). DGP model was established by intraperitoneal injection of streptozotocin (STZ, 2%), and raised with high-sugar high-fat diet irregularly. EA (sparse-dense, 10 Hz/50 Hz, 2 mA, 20 min) was applied at "Zusanli" (ST 36), "Liangmen" (ST 21) and "Sanyinjiao" (SP 6), and the corresponding non-acupoints of the 3 acupoints, daily for 15 days. The rats in metoclopramide group received intragastric administration of metoclopramide (1.7%, 1 mL/100 g) for 15 days, once a day. Blood sugar was determined with One Touch blood glucose test paper. The gastric emptying rate (GER) and the intestinal propulsion rate (IPR) were measured by intragastric phenol red. The ultrastructure of ICC was detected by transmission electron microscopy. The expression levels of c-kit receptor protein and SCF mRNA of gastric antrum were examined respectively by Western blot and RT-PCR. RESULTS: Compared with the normal group, the blood glucose significantly increased in the model group (P<0.01), while the GER, IRP and the expression level of SCF mRNA in the gastric antrum significantly decreased (P<0.01), and the ultrastructure of ICC appeared apoptosis-like changes. The blood glucose of the EA group was obviously decreased compared with that of the model group (P<0.05); the GER and IRP significantly increased(P<0.05, P<0.01); the expression level of SCF mRNA increased (P<0.01), the number of ICC increased and its ultrastructure was repaired. There was some relief on ICC ultrastructure in the acupoint group compared with that in the non-acupoint group; and SCF mRNA increased (P<0.05). There was no significant difference on c-kit receptor expression among all the modeling groups (P＞0.05). CONCLUSIONS: EA at ST 36, etc. can regulate the blood glucose and improve gastrointestinal emptying in DGP rats. The mechanism may be related to up-regulating SCF mRNA, repairing ICC ultrastructure, restoring the pacing function, and improving gastrointestinal motility.
ABSTRACT
Objective Resistin,also known as an adipose tissue-specific secretory factor,is involved in the development and progression of artherosclerosis by inducing dysfunction of endothelial cells,but its action mechanism has been rarely studied.The purpose of this study was to investigate the effect of resistin on the function of human coronary artery endothelial cells (HCAECs) and the underlying mechanisms.Methods We constructed the lentiviral vasodilator-stimulated phosphoprotein (VASP) shRNA interfering sequence (the LV-siVASP group) and a negative control vector (the LV-sicntr group),transfected HCAECs with VASP siRNA or control siRNA,and determined the VASP mRNA and protein expressions by quantitative RT-PCR and Western blot,respectively.We treated the HCAECs with resistin at the concentrations of 0,10,50,100,and 200 ng/mL followed by-detection of its effects on the proliferation and migration of the cells by MTT and Transwell chamber assay,respectively.We made comparisons between the LV-siVASP and LV-sicntr groups in the proliferation and migration of the HCAECs treated with 100 ng/mL resistin,the expression of vascular endothelial growth factor receptor-2 (VEGFR2) by immunofluorescent staining,and the activity of RhoA by pull-down assays.Results Compared with the LV-sicntr group,the LV-siVASP group showed significantly inhibited expressions of RNA (100％ vs [68.1±0.8]％,P＜0.05) and the VASP protein (100％ vs [59.3± 1.7] ％,P＜0.05) Treatment with resistin at 50-200 ng/mL markedly increased the proliferation of the HCAECs at 48 hours (P＜0.05) and induced a dose-dependent promotion of their migration at 24 hours (P＜0.05).No significant difference was observed in the expression VEGFR2 between the two groups,while the activity if RhoA was remarkably reduced in the LV-siVASP group ([41.3±3.1] ％) as compared with the LV-sicntr group (P＜0.05).Conclusion Resistin promotes the proliferation and migration of HCAECs,and the influence of VASP ablation on the action of resistin is associated with the decreased activity of RhoA.
ABSTRACT
The market and literature were studied to understand the existing situation of Magnoliae Officinalis Cortex goods, and the collected samples were analyzed, combined with the actual production, a new standard of Magnoliae Officinalis Cortex commercial specification and grade was drafted. Magnoliae Officinalis Cortex goods was divided into two categories according to the source in the old standard. Then each category was divided into four kinds of specifications according to the site. Each kind of specification was divided into several grades according to the length and weight. To judge the quality of Magnoliae Officinalis Cortex goods was mainly based on the appearance quality. In the new standard, the classification of commercial specification and grade is based on the thickness, magnolol and honokiol content. The goods of Magnoliae Officinalis Cortex can be divided into three specifications: Tongpu, Genpu and Doupu. Tongpu is divided into three grades, the remaining two are not graded.
Subject(s)
Magnolia , ChemistryABSTRACT
OBJECTIVE@#To establish stable and controllable brain injury with accurate degree and good repeatability in rat model.@*METHODS@#Controlled cortical impact (CCI) device was used to prepare for the rat brain injury model by the impact head of different model (Group A No. 4, Group B No. 5, Group C No. 6) and the impact depth (Group A: 1.5-2.0 mm, Group B: 2.5-3.0 mm, Group C: 3.5-4.0 mm) with impact time of 0.1 s and impact velocity of 2.5 m/s. Twelve rats with three months of age were used in each group (the impact depth of every two rats was added 1 mm respectively). After modeling for 1 h, magnetic resonance imaging (MRI) was received and brain histopathology was observed to assess degree of injury by model parameters of three groups.@*RESULTS@#After modeling of Group A, MRI showed that the cortex structure was damaged with a small amount of bleeding in center and mild edema around, and the total volume of injury was (28.69±4.94) mm(3). Pathology revealed the injury was confined to the superficial cortical with mild edema of nerve cell, which was assessed as mild cerebral contusion. While after modeling, MRI of Group B showed that the structure of cortex and medulla were damaged simultaneously and extended to cerebral nuclei zone, with 4 cases of hematoma in the center and larger edema range around, and the total volume of injury was (78.38±9.28) mm(3). Pathology revealed the injury range was reached nuclei zone, with swell of nerve cell and mitochondria, which was assessed to moderate cerebral contusion. After modeling of Group C, MRI showed that extensive tissue injury was appeared in cortex and medulla and deep nuclei, with 9 cases of hematoma and large edema signal of surrounding tissue T2WI, while in 5 cases, lateral nucleus of injury signal was increased, and the total volume of injury was (135.89±24.80) mm(3). Pathology revealed the deep cerebral nuclei was damaged, with the disappearance of neuronal structure and vacuolization of mitochondria, which was assessed as severe cerebral contusion. MRI changes were consistent with pathological changes in three groups of model, and the injury range was significantly different (P<0.01).@*CONCLUSIONS@#Application of CCI can make stable quantitative traumatic brain injury model, which overcomes the randomness in previous injury model and possesses highly unity in iconography and pathology changes. This can provide quantitative modeling reference for clinical research.
ABSTRACT
Objective: To establish stable and controllable brain injury with accurate degree and good repeatability in rat model. Methods: Controlled cortical impact (CCI) device was used to prepare for the rat brain injury model by the impact head of different model (Group A No. 4, Group B No. 5, Group C No. 6) and the impact depth (Group A: 1.5-2.0 mm, Group B: 2.5-3.0 mm, Group C: 3.5-4.0 mm) with impact time of 0.1 s and impact velocity of 2.5 m/s. Twelve rats with three months of age were used in each group (the impact depth of every two rats was added 1 mm respectively). After modeling for 1 h, magnetic resonance imaging (MRI) was received and brain histopathology was observed to assess degree of injury by model parameters of three groups. Results: After modeling of Group A, MRI showed that the cortex structure was damaged with a small amount of bleeding in center and mild edema around, and the total volume of injury was (28.69±4.94) mm