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Objective: To observe the effect of electroacupuncture (EA) of "Zusanli" (ST36), "Sanyinjiao" (SP6) and "Liangmen" (ST21) on gastrointestinal motility, blood glucose content and expression of autophagy-related proteins 1 light chain 3 (LC3), p62, phosphatidyli-nositol-3 kinase (PI3K), protein kinase B (Akt), p-Akt and mammalian target protein of rapamycin (mTOR) of interstitial cells of Cajal (ICCs) in the cultured gastric antrum cells in diabetic gastroparesis (DGP) rats, so as to reveal its mechanisms underlying improvement of DGP., Methods: A total of 45 Sprague Dawley (SD) rats were randomly divided into blank control, model, EA, medication (3-methyladenine, 3-MA) and EA+3-MA groups, with 9 rats in each group. The DGP model was established by intraperitoneal injection of 2% streptozotocin (STZ) combined with high-fat and high sugar diet for 8 weeks. The gastric emptying rate was measured by using gavage of phenol red (to measure the propelling length of the phenol red/total length of small intestine ×100%). The symptom score (mental state, coat color and luster, behavior and activity, stool traits) of rats was observed every week and the blood glucose content was measured by using a glucometer. EA (20 Hz/100 Hz, 2 mA) was applied to unilateral ST36, SP6 and ST21 alternatively for 15 min, once daily, 5 days a week for 3 weeks. Rats of the 3-MA and 3-MA+EA groups received intraperitoneal injection of 3-MA (30 mg·kg -1 ·d -1 , 10 mg/mL), once daily, 5 days a week for 3 weeks. After 15 days' intervention, the rats were operated for gastric emptying rate test, specimen collection, isolation, and culture of primary ICCs. The expression levels of microtubule associated protein LC3, p62, PI3K, Akt, p-Akt and mTOR of ICCs of cultured gastric antrum cells were detected using Western blot, and the number of autophagosomes in ICC of gastric antrum was observed under transmission electron microscope., Results: Compared with the blank control group, the symptom score, blood glucose, and the expression levels of p62, class Ⅰ PI3K, Akt, p-Akt and mTOR proteins were increased significantly ( P <0.01), while the gastric emptying rate and ratio of LC3Ⅱ/LC3Ⅰ and the expression level of class Ⅲ PI3K protein were significantly decreased ( P <0.05, P <0.01) in the model group. In comparison with the model group, the increase of symptom score, blood glucose, and expression levels of p62, class Ⅰ PI3K, Akt, p-Akt and mTOR proteins and the decrease of gastric empty rate and LC3Ⅱ/LC3Ⅰ ratio and the expression level of class Ⅲ PI3K protein were all reversed in both EA and EA+3-MA groups ( P <0.05, P <0.01), rather than in the 3-MA group. In addition, 3-MA also reversed modeling-induced increase of class Ⅰ PI3K, Akt, p-Akt and mTOR proteins expression ( P <0.01). No significant differences were found between the EA and EA+3-MA in downregulating the levels of symptom score and blood glucose content, and in upregulating gastric empty rate( P >0.05). The effect of EA was notably superior to that of EA+3-MA in upregulating the ratio of LC3Ⅱ/LC3Ⅰ and the expression level of class Ⅲ PI3K protein, and in downregulating the expression of p62, class Ⅰ PI3K, Akt, p-Akt and mTOR proteins ( P <0.05, P <0.01). The findings of transmission electron microscopy showed obvious swelling, breakage of some mitochondrial cristae in the ICC cells of antrum and no autophagosomes in the model group and 3-MA group, which was milder in the damage of mitochondrial cristae and marked increase in the autophagosomes in both EA and EA+3-MA groups., Conclusion: EA can improve the gastrointestinal motility and symptoms in DGP rats, which may be related to its functions in downregulating PI3K/Akt/mTOR signaling to promote autophagy level of ICC.

Objective: To observe the effect of electroacupuncture (EA) combined with Zhuang-medicine-thread moxibustion on expression of apoptosis-related factors in gastric antrum of diabetic gastroparesis (DGP) rats, so as to explore its mechanism underlying improvement of DGP., Methods: Male SD rats were randomly divided into normal, model, medication, EA, Zhuang-medicine-thread moxibustion (moxibustion) and EA+moxibustion (combination) groups (12 rats in each group). The DGP model was established by intraperitoneal injection of streptozotocin (STZ). Rats of the medication group were treated by gavage of 0.15 mg/mL mosapride citrate suspension （10 mL/kg）. EA (10 Hz/50 Hz, 2 mA, 20 min) or Zhuang-medicine-thread moxibustion (3 cones) was applied to "Zhongwan" (CV12), bilateral "Neiguan" (PC6) and bilateral "Sanyinjiao" (SP6) of the related groups, once a day for 3 weeks. The blood glucose, gastric emptying rate and intestinal propulsion rate of rats were measured. The apoptosis index of gastric antrum cells were observed by TUNEL staining. The protein and mRNA expressions of Caspase-3, B-cell lymphoma/leukemia-2 (Bcl-2) and Bcl-2 associated X protein (Bax) in gastric antrum were detected by Wes-tern blot and real-time quantitative PCR, respectively., Results: Compared with the normal group, the blood glucose, the apoptosis index, the protein and gene expressions of Caspase-3 and Bax were significantly increased ( P <0.01), and the gastric emptying rate, intestinal propulsive rate, the protein and gene expressions of Bcl-2 were considerably decreased ( P <0.01) in the model group. In contrast to the model group, the blood glucose in the EA, moxibustion and combination groups, the apoptosis index in the 4 treatment groups, as well as Caspase-3 protein, Bax protein and mRNA expressions in the medication, EA and combination groups, Caspase-3 protein and mRNA, Bax mRNA expressions in the moxibustion group were significantly decreased ( P <0.01, P <0.05); while the gastric emptying rate and intestinal propulsive rate in the 4 treatment groups, and Bcl-2 protein and mRNA expressions in the medication and combination groups, Bcl-2 mRNA expressions in the EA and moxibustion groups were obviously increased ( P <0.01). The effects of EA+moxibustion were significantly superior to those of simple EA, moxibustion or medication in increasing gastric emptying rate and intestinal propulsive rate, and in lowering blood glucose ( P <0.05, P <0.01). And the effects of the combination treatment were better than those of EA in lowering Caspase-3 protein and Bax mRNA expressions ( P <0.01), and in increasing Bcl-2 protein and mRNA expressions ( P <0.05, P <0.01). Also the effects of the combination treatment were better than those of moxibustion in lowering the apoptosis index, Caspase-3 protein, and Bax protein and mRNA expressions ( P <0.01, P <0.05), and in increasing Bcl-2 protein expression ( P <0.05)., Conclusion: EA combined with Zhuang-medicine-thread moxibustion can reduce blood glucose and improve gastrointestinal motility in DGP rats, which may be related to its effect in regulating of Caspase-3, Bax and Bcl-2 expression.

Objective: To observe the effect of point-moxibustion and electroacupuncture(EA) on gastrointestinal motility, and expression of endothelial nitric oxide synthase(eNOS) mRNA and angiotensin 2(AT Ⅱ) mRNA in gastric antrum in diabetic gastroparesis(DGP) rats, so as to reveal the different effect between point-moxibustion and EA, and explore the mechanism underlying improvement of DGP., Methods: Forty SD rats were randomly divided into normal, model, point-moxibustion and EA groups, 10 rats in each group. The DGP model was established by intraperitoneal injection of streptozotocin (STZ, 2%, 55 mmol/kg) and fed with high-sugar and high-fat fodder. "Zusanli" (ST 36), "Sanyinjiao" (SP 6) and "Liangmen" (ST 21) were chosen to be stimulated with moxibustion at 0, 10 and 20 min in the point-moxibustion group or with EA for 20 min in the EA group. The treatment was conducted once daily for 15 days. After treatment, blood glucose, gastric emptying rate and small intestinal propulsive rate were measured. The plasma endothelin 1(ET-1) content was detected by ELISA. The mRNA expression of eNOS and AT Ⅱ in gastric antrum were detected by Real-time PCR., Results: Compared with the normal group, the blood glucose in the model group was significantly higher ( P <0.01). Compared with the model group, the blood glucose decreased significantly in the point-moxibustion group and EA group ( P <0.05). In comparison with the normal group, gastric emptying rate and small intestinal propulsive rate were decreased, the content of plasma ET-1 was increased, the expression of eNOS mRNA in gastric antrum was significantly reduced and the expression of ATⅡ mRNA was significantly increased in the model group( P <0.01). In comparison with the model group, gastric emptying rate and small intestinal propulsive rate were increased, the content of plasma ET-1 was decreased, the expression of eNOS mRNA in gastric antrum was higher and the expression of AT Ⅱ mRNA was significantly lower in both point-moxibustion group and EA group ( P <0.05), and there were no significant differences between the two treatment groups in above mentioned indexes ( P >0.05)., Conclusions: Both point-moxibustion and EA can effectively promote gastrointestinal motility in DGP rats and improve the symptoms of delayed gastric emptying. The two therapies are effective, and their mechanism may be related to the increase of eNOS mRNA expression and the decrease of AT Ⅱ mRNA expression in gastric antrum.

Objective: To observe the effect of electroacupuncture (EA) stimulation of "Zusanli" (ST 36), "Liangmen" (ST 21) and "Sanyinjiao" (SP 6) on gastrointestinal motility and expression of ghrelin and growth hormone secretagogue receptor (GHSR) mRNA in diabetic gastroparesis (diabetic gastroparesis, DGP) rats, so as to explore its mechanism underlying improvement of diabetic gastroparesis., Methods: Sixty SD rats were randomly and equally divided into normal control, model, EA acupoint, EA non-acupoint, medication (metoclopramide) groups (n = 12 rat/group). The DGP model was established by intraperitoneal injection of streptozotocin (STZ, 55 mg/kg, 2%) and raised by high-sugar and high-fat fodder irregularly for 8 weeks. EA was applied to "Zusanli" (ST 36), "Liangmen" (ST 21) and "Sanyinjiao" (SP 6), and non-acupoints (about 5 mm lateral to the 3 acupoints, respectively). Those rats of the medication group were given with metoclopramide (1 mL/100 g). The blood sugar level was measured by glucometer, and the urine sugar level measured by Tes-Tape [urine glucose test paper]. The intestinal propulsion rate (IPR) was identified as ratio of the phenolsulfonphthalein promoting distance and the total length of the small intestine, and the gastric emptying rate (GER) was assessed by gavage of a liquid test meal (1.5 mL) containing a nonabsorbable marker (0. 5 mg/mL of phenol red in 5% glucose solution) and identification after being sacrificed 72 h after modeling. The expression levels of ghrelin mRNA and GHSR mRNA of gastric antrum tissue were examined by Quantitative real-time PCR., Results: In comparison with the normal control group, the blood sugar and urine sugar levels of the model group was significantly increased in the model group (P < 0.01), while the GER and IPR and the expression level of ghrelin mRNA and GHSR mRNA in the gastric antrum were significantly decreased in the model group (P < 0.01, P < 0.05). Following EA stimulation of acupoint, the blood sugar and urine sugar levels were obviously down-regulated (P < 0.05), both GER and IPR, and ghrelin mRNA and GHSR mRNA expression levels in the EA-acupoint group were significantly up-regulated (P < 0.05, P < 0.01). No significant changes were found in both blood sugar and urine sugar levels, in both GER and IPR, and ghrelin mRNA and GHSR mRNA expression levels of both EA non-acupoint and medication groups after the intervention (P > 0.05). The GER and IPR were significantly higher in the EA acupoint group than in the EA non-acupoint group (P < 0.05, P < 0.01)., Conclusion: EA stimulation of ST 36, ST 21 and SP 6 can promote the gastrointestinal motility of DGP rats, which may be closely associated with its effects in up-regulating the expression level of ghrelin mRNA and GHSR mRNA in gastric antrum.

Objective: To observe the effect of Zhuang-medicine medicated-thread moxibustion therapy on interstitial cells of Cajal (ICC) in the gastric antrum of diabetic gastroparesis (DGP) rats, so as to investigate its mechanism underlying improving DGP., Methods: Sprague-Dawley (SD) male rats were randomly divided into 3 groups: control (n = 30) , model (n = 30) and moxibustion (n = 30). The DGP model was established by intraperitoneal injection of streptozotocin (STZ) and by feeding the rats with high fat-sugar forage. Zhuang-medicine medicated-thread moxibustion was applied to "Zhongwan" (CV 12), bilateral "Pishu" (BL 20), "Weishu" (BL 21), "Neiguan" (PC 6) and "Zusanli" (ST 36) once per day, for 3 weeks except weekends. The gastrointestinal propulsion rate and weight of stool in 24 h were determined, and c-kit (a marker for ICC) expression of the gastric antrum tissue was measured by immunohistochemistry., Results: The stool weight was significantly higher in the model group than in the control group (P < 0.01), but the rate of gastrointestinal propulsion and the rate of c-kit immunoreaction (IR) positive cells in the gastric antrum tissue were significantly lower in the model group than in the control group (P < 0.01). After moxibustion, the increased stool weight and the decreased gastrointestinal propulsion rate and decreased c-kit IR-positive cell rate were reversed (P < 0.01)., Conclusion: Zhuang-medicine medicated-thread moxibustion therapy can improve gastrointestinal function in DGP rats, which may be associated with its effect in up-regulating the expression of c-kit IR-positive ICC.

The aim of the present study was to explore the role of orphan G protein-coupled receptor 55 (GPR55) in diabetic gastroparesis (DG). Streptozotocin (STZ) was used to mimic the DG model, and the body weight and blood glucose concentration were tested 4 weeks after STZ injection (i.p.). Electrogastrogram and phenolsulfonphthalein test were used for detecting gastric emptying. Motilin (MTL), gastrin (GAS), vasoactive intestinal peptide (VIP), and somatostatin (SS) levels in plasma were determined using radioimmunology. Real-time PCR and Western blot were applied to identify the expression of GPR55 in gastric tissue, and immunohistochemistry was used to detect the distribution. The effect of lysophosphatidylinositol (LPI), an agonist of GPR55, was observed. STZ mice showed increased blood glucose concentration, lower body weight, decreased amplitude of slow wave, and delayed gastric emptying. LPI antagonized these effects of STZ. Compared to the control group, STZ caused significant decreases of MTL and GAS levels (P < 0.01), as well as increases of SS and VIP levels (P < 0.01). The changes of these hormones induced by STZ were counteracted when using LPI. GPR55 located in mice stomach, and it was up-regulated in DG. Although LPI showed no effects on the distribution and expression of GPR55 in normal mice, it could inhibit STZ-induced GPR55 up-regulation. These results suggest GPR55 is involved in the regulation of gastric movement of DG, and may serve as a new target of DG treatment. LPI, an agonist of GPR55, can protect against STZ-induced DG, and the mechanism may involve the change of GPR55 expression and modification of gastrointestinal movement regulating hormones.

Objective: To observe the effects of Tangweian granule on 5-HT(2A)R in rat model with diabetic gastroparesis (DGP)., Method: The rats with diabetic gastroparesis induced by injecting alloxan and giving 200% Radix Rehmanniae preparata were divided into four groups randomly: Tangweian high dosage group, Tangweian low dosage group, motilium control group and the model control group, 10 rats each group. Each group was irrigated with drugs during establishing the model. Additionally, we chose 10 rats by way of normal control group. Further more, Tangweian high dosage group were irrigated stomach with gliclazide 20 mg x kg(-1) and Tangweian granule 31.75 g x kg(-1); Tangweian low dosage group were irrigated stomach with gliclazide 20 mg x kg(-1) and Tangweian granule 15.88 g x kg(-1); motilium control group were irrigated stomach with gliclazide 20 mg x kg(-1) and motilium 3.75 mg x kg(-1) and the model control group were irrigated stomach with distilled water. Then the effects of Tangweian granule on 5-HT(2A)R were observed., Result: The curative group had better effects than the control group in lowering the blood sugar and the level of 5-HT(2A)R content (P < 0.01). And there was significant difference between the curative group and control group (P < 0.05)., Conclusion: It is verified that Tangweian granule has obvious effects on lowering the blood sugar and improving the level of 5-HT(2A)R.