Ji Hu

Promoted Interaction of Nuclear Factor-κB with Demethylated Cystathionine-β-Synthetase Gene Contributes to Gastric Hypersensitivity in Diabetic Rats

Patients with long-standing diabetes frequently demonstrate gastric hypersensitivity with an unknown mechanism. The present study was designed to investigate roles for nuclear factor-κB (NF-κB) and the endogenous H2S-producing enzyme cystathionine-β-synthetase (CBS) signaling pathways by examining cbs gene methylation status in adult rats with diabetes. Intraperitoneal injection of streptozotocin (STZ) produced gastric hypersensitivity in female rats in response to gastric balloon distention. Treatment with the CBS inhibitor aminooxyacetic acid significantly attenuated STZ-induced gastric hypersensitivity in a dose-dependent fashion. Aminooxyacetic acid treatment also reversed hyperexcitability of gastric-specific dorsal root ganglion (DRG) neurons labeled by the dye DiI in diabetic rats. Conversely, the H2S donor NaHS enhanced neuronal excitability of gastric DRG neurons. Expression of CBS and p65 were markedly enhanced in gastric DRGs in diabetic rats. Blockade of NF-κB signaling using pyrrolidine dithiocarbamate reversed the upregulation of CBS expression. Interestingly, STZ treatment led to a significant demethylation of CpG islands in the cbs gene promoter region, as determined by methylation-specific PCR and bisulfite sequencing. STZ treatment also remarkably downregulated the expression of DNA methyltransferase 3a and 3b. More importantly, STZ treatment significantly enhanced the ability of cbs to bind DNA at the p65 consensus site, as shown by chromatin immunoprecipitation assays. Our findings suggest that upregulation of cbs expression is attributed to cbs promoter DNA demethylation and p65 activation and that the enhanced interaction of the cbs gene and p65 contributes to gastric hypersensitivity in diabetes. This finding may guide the development and evaluation of new treatment modalities for patients with diabetic gastric hypersensitivity.

Promoted Interaction of Nuclear Factor-κB With Demethylated Purinergic P2X3 Receptor Gene Contributes to Neuropathic Pain in Rats With Diabetes

Painful diabetic neuropathy is a common complication of diabetes produced by mechanisms that as yet are incompletely defined. The aim of this study was to investigate the roles of nuclear factor-κB (NF-κB) in the regulation of purinergic receptor P2X ligand-gated ion channel 3 (P2X3R) plasticity in dorsal root ganglion (DRG) neurons of rats with painful diabetes. Here, we showed that hindpaw pain hypersensitivity in streptozocin-induced diabetic rats was attenuated by treatment with purinergic receptor antagonist suramin or A-317491. The expression and function of P2X3Rs was markedly enhanced in hindpaw-innervated DRG neurons in diabetic rats. The CpG (cytosine guanine dinucleotide) island in the p2x3r gene promoter region was significantly demethylated, and the expression of DNA methyltransferase 3b was remarkably downregulated in DRGs in diabetic rats. The binding ability of p65 (an active form of NF-κB) with the p2x3r gene promoter region and p65 expression were enhanced significantly in diabetes. The inhibition of p65 signaling using the NF-κB inhibitor pyrrolidine dithiocarbamate or recombinant lentiviral vectors designated as lentiviral vector-p65 small interfering RNA remarkably suppressed P2X3R activities and attenuated diabetic pain hypersensitivity. Insulin treatment significantly attenuated pain hypersensitivity and suppressed the expression of p65 and P2X3Rs. Our findings suggest that the p2x3r gene promoter DNA demethylation and enhanced interaction with p65 contributes to P2X3R sensitization and diabetic pain hypersensitivity.

Colonic Hypersensitivity and Sensitization of Voltage-gated Sodium Channels in Primary Sensory Neurons in Rats with Diabetes.

Background/Aims Patients with long-standing diabetes often demonstrate intestinal dysfunction and abdominal pain. However, the pathophysiology of abdominal pain in diabetic patients remains elusive. The purpose of study was to determine roles of voltage-gated sodium channels in dorsal root ganglion (DRG) in colonic hypersensitivity of rats with diabetes. Methods Diabetic models were induced by a single intraperitoneal injection of streptozotocin (STZ; 65 mg/kg) in adult female rats, while the control rats received citrate buffer only. Behavioral responses to colorectal distention were used to determine colonic sensitivity in rats. Colon projection DRG neurons labeled with DiI were acutely dissociated for measuring excitability and sodium channel currents by whole-cell patch clamp recordings. Western blot analysis was employed to measure the expression of NaV1.7 and NaV1.8 of colon DRGs. Results STZ injection produced a significantly lower distention threshold than control rats in responding to colorectal distention. STZ injection also depolarized the resting membrane potentials, hyperpolarized action potential threshold, decreased rheobase and increased frequency of action potentials evoked by 2 and 3 times rheobase and ramp current stimulation. Furthermore, STZ injection enhanced neuronal sodium current densities of DRG neurons innervating the colon. STZ injection also led to a significant upregulation of NaV1.7 and NaV1.8 expression in colon DRGs compared with age and sex-matched control rats. Conclusions Our results suggest that enhanced neuronal excitability following STZ injection, which may be mediated by upregulation of NaV1.7 and NaV1.8 expression in DRGs, may play an important role in colonic hypersensitivity in rats with diabetes.

Alpha-lipoic Acid suppresses P2X receptor activities and visceral hypersensitivity to colorectal distention in diabetic rats

The present study was designed to investigate the roles of P2X3 receptors in dorsal root ganglion (DRG) neurons in colonic hypersensitivity and the effects of alpha-lipoic acid (ALA) on P2X3 receptor activity and colonic hypersensitivity of diabetic rats. Streptozotocin (STZ) was used to induce diabetic model. Abdominal withdrawal reflex (AWR) responding to colorectal distention (CRD) was recorded as colonic sensitivity. ATP-induced current density of colon-specific DRG (T13-L2 DRGs) neurons was measured with whole-cell patch clamp. The expression of P2X3Rs of T13-L2 DRGs was measured by western blot analysis. The results showed that AWR scores significantly increased after STZ injection. P2X3R expression and ATP current density of T13-L2 DRG neurons were enhanced in diabetic rats. Intraperitoneal injection with ALA once a day for 1 week remarkably reduced P2X3R expression and ATP current density in diabetic rats. Importantly, ALA treatment attenuated colonic hypersensitivity in diabetic rats. Our data suggest that STZ injection increases expression and function of P2X3 receptors of colon-specific DRG neurons, thus contributing to colonic hypersensitivity in diabetic rats. Administration of ALA attenuates diabetic colonic hypersensitivity, which is most likely mediated by suppressing expression and function of P2X3 receptors in DRGs of diabetic rats.

α-lipoic acid suppresses neuronal excitability and attenuates colonic hypersensitivity to colorectal distention in diabetic rats

Aim Patients with long-standing diabetes often demonstrate intestinal dysfunction, characterized as constipation or colonic hypersensitivity. Our previous studies have demonstrated the roles of voltage-gated sodium channels NaV1.7 and NaV1.8 in dorsal root ganglion (DRG) in colonic hypersensitivity of rats with diabetes. This study was designed to determine roles of antioxidant α-lipoic acid (ALA) on sodium channel activities and colonic hypersensitivity of rats with diabetes. Methods Streptozotocin was used to induce diabetes in adult female rats. Colonic sensitivity was measured by behavioral responses to colorectal distention in rats. The excitability and sodium channel currents of colon projection DRG neurons labeled with DiI were measured by whole-cell patch-clamp recordings. The expressions of NaV1.7 and NaV1.8 of colon DRGs were measured by western blot analysis. Results ALA treatment significantly increased distention threshold in responding to colorectal distension in diabetic rats compared with normal saline treatment. ALA treatment also hyper-polarized the resting membrane potentials, depolarized action potential threshold, increased rheobase, and decreased frequency of action potentials evoked by ramp current stimulation. Furthermore, ALA treatment also reduced neuronal sodium current densities of DRG neurons innervating the colon from rats with diabetes. In addition, ALA treatment significantly downregulated NaV1.7 and NaV1.8 expression in colon DRGs from rats with diabetes. Conclusion Our results suggest that ALA plays an analgesic role, which was likely mediated by downregulation of NaV1.7 and NaV1.8 expressions and functions, thus providing experimental evidence for using ALA to treat colonic hypersensitivity in patients with diabetic visceral pain.

Clinical characteristics and risk factors of diabetic peripheral neuropathy of type 1 diabetes mellitus patients

BACKGROUND Small nerve fibers are more easily injured than large ones for diabetic peripheral neuropathy (DPN). The study investigated the characteristics and related risk factors of DPN of T1DM patients using nerve conduction velocity and CPT values, which provided evidences for its early diagnosis. METHODS 70 T1DM patients and 48 healthy volunteers were included. All subjects accepted nerve conduction velocity and CPT examinations for four limbs. Detailed clinical indicators were recorded. CPT values were compared between TIDM group and control group. The risk factors affecting DPN were further explored. RESULTS Compared with the control group, CPT values under three frequencies were decreased in T1DM group. The abnormality rate of sural nerves was higher than that of median nerves (P<0.001). Median nerve dysfunction mainly presented as hypoesthesia under 250Hz and 5Hz current stimulus. And sural nerve dysfunction mainly presented as hyperesthesia under three frequencies. Compared with left median nerve, abnormal rate of right median nerve was higher under 2000Hz current stimulus (P=0.035). However, abnormal rate of left sural nerve was higher than that of right side under 250Hz and 5Hz current stimulus (P=0.001, <0.001). Duration, NDS scores and CPT values of right median nerve under 2000Hz current stimulus were independent risk factors of abnormal nerve conduction velocity. CONCLUSIONS The study proved that DPN of T1DM are mainly lower limb-injured., amyelinated and thin myelinated nerve fiber-involved. CPT can be combined with traditional nerve conduction velocity examination, which will help the diagnosis of DPN of T1DM earlier and more comprehensively.

The Association Between Fasting C-peptide and Gastrointestinal Symptoms of Gastroparesis in Type 2 Diabetic Patients

Background/Aims The relationship between C-peptide levels and gastrointestinal (GI) symptoms in type 2 diabetic patients is not clear. The purpose of this study is to examine the association between fasting C-peptide and GI symptoms of gastroparesis in type 2 diabetic patients. Methods We recruited 333 type 2 diabetic patients into the present study. All patients filled out questionnaires of gastroparesis cardinal symptom index (GCSI) to evaluate GI symptoms. Hospital anxiety and depression scale were adopted to define anxiety and depression. Patients with GCSI scores ≥ 1.9 were regarded as having symptoms of gastroparesis. Results In our study, 71 (21.3%) type 2 diabetic patients had GCSI scores ≥ 1.9. In comparison to patients with scores < 1.9, those with scores ≥ 1.9 had significantly lower fasting c-peptide levels (1.49 ng/mL vs 1.94 ng/mL, P < 0.001), higher prevalence of depression (40.9% vs 18.3%, P < 0.001) and anxiety (28.2% vs 13.0%, P = 0.002). Multivariate logistic regression revealed that fasting C-peptide was still significantly associated with symptoms of gastroparesis (odds ratio, 0.67; 95% confidence intervals, 0.48–0.94; P = 0.021), even after adjustments for age, sex, body mass index, HbA1c, current smoking and drinking status, anxiety, and depression. Furthermore, linear regressions showed that fasting C-peptide was independently and negatively related to GCSI scores (standardized regression coefficient, −0.29; P < 0.001) in patients with at least one GI symptom. Conclusion GI symptoms of diabetic gastroparesis affect approximately 20% of type 2 diabetes patients and are associated with lower fasting C-peptide levels independent of depression and anxiety status.

Treatment of diabetic gastroparesis with botulinum toxin injection guided by endoscopic ultrasound in a patient with type 1 diabetes: the first report

Diabetic gastroparesis is a well-recognized diabetic automatic neuropathy characterized by the delay of gastric emptying without gastric outlet obstruction. Upper gastrointestinal symptoms have been reported by up to 19 % of diabetic patients, and 48–65 % of these patients have been shown to have diabetic gastroparesis [1]. The cardinal symptoms include postprandial fullness, nausea, vomiting, and bloating. These symptoms caused by diabetic gastroparesis may result in malnutrition, malabsorbtion, impaired glucose control, and a poor quality of life. Patients with refractory diabetic gastroparesis who experience severe symptoms are not able to maintain sufficient nutritional intake and easily cause ketosis. Previous study revealed that diabetic gastroparesis was associated with higher mortality, morbidity, increased hospitalizations, emergency department, and doctor visits [2]. Primary treatment for diabetic gastroparesis includes correction of exacerbating factors, nutritional support, pharmacologic therapy, and surgical therapy. However, these options for patients have limitations such as high recurrence rate and various side effects. Botulinum toxin A injection is an alternative option for the treatment of diabetic gastroparesis by reducing intrapyloric muscular tone. Botulinum toxin A injection into the pyloric muscle was usually guided by endoscopies in previous studies. The limitations of endoscopic botulinum toxin A injection include uncertain depth of injection and the potential risk of perforation. Here, we present a case of intrapyloric botulinum toxin A injection guided by endoscopic ultrasound (EUS) in a patient with diabetic gastroparesis.

Soluble B7-H3 (sB7-H3) is over-expressed in the serum of type 1 diabetes patients

Type 1 Diabetes (T1D) is an autoimmune disease resulting from insulin-secreting β-cells mediated by autoreactive T cells. We demonstrated increased level of sB7-H3 in T1D patients than in healthy control group. This result suggests that B7-H3 may be may be a promising biomarker associated with the pathogenesis of T1D.

The honeymoon period repeatedly appears in two cases of type 1 diabetes mellitus

Type 1 diabetes mellitus (T1DM) is a chronic autoimmune disease caused by beta cell destruction resulting from interactions of genetic, environment, and immunologic factors, which is characterized by the complete or near total deficiency of insulin and being positive in autoantibodies. Honeymoon phase, a natural phenomenon, occurs in newly diagnosed type 1 diabetes patients, starting after the usage of insulin treatment and during which patients require few exogenous insulin to maintain the normal range of blood glucose and have HbA1c less than 7% [1]. The duration of honeymoon usually ranges from 1 month to 13 years [2]. In the previous clinical research, most patients have the honeymoon period after insulin treatment, and no observation has described a case in which the honeymoon period appears twice. Here, we present two cases that were diagnosed with T1DM and then underwent more than one honeymoon periods. Case presentation