Aerobic training with moderate or high doses of vitamin D improve liver enzymes, LXRα and PGC-1α levels in rats with T2DM.

Dysregulation of key transcription factors involved in hepatic energy metabolism, such as peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1α) and liver X receptor alpha (LXRα), has been observed in T2DM. The present study aims to investigate the effects of aerobic training and vitamin D supplementation on liver enzyme levels and the levels of PGC-1α and LXRα proteins in hepatocytes, in a rat model of T2DM. The study involved 56 male Wistar rats, divided into two groups: one was non-diabetic and acted as a control group (n = 8), and the other had induced diabetes (n = 48). The diabetic rats were then split into six subgroups: two groups received high or moderate doses of vitamin D and aerobic training (D + AT + HD and D + AT + MD); two groups received high or moderate doses of vitamin D alone (D + HD and D + MD); one group underwent aerobic training with vehicle (sesame oil; D + AT + oil), and one group was a diabetic control receiving only sesame oil (oil-receiving). The D + AT + HD and D + HD groups received 10,000 IU of vitamin D, while the D + AT + MD and D + MD groups received 5000 IU of vitamin D once a week by injection. The D + AT + oil group and the sham group received sesame oil. After eight weeks of treatment, body weight, BMI, food intake, serum insulin, glucose, 25-hydroxyvitamin D, ALT, AST, and visceral fat were measured. The levels of PGC-1α and LXRα proteins in the liver was assessed by western blotting. Statistical analysis was performed using the paired t-test, one-way analysis of variance (ANOVA), and the Tukey post hoc test at a significance level of P < 0.05. Body weight, food intake, and BMI decreased significantly in the D + AT + HD, D + AT + MD, D + AT + oil, D + HD, and D + MD groups with the highest reduction being observed in body weight and BMI in the D + AT + HD group. The D + AT + HD group exhibited the lowest levels of insulin, glucose, and HOMA-IR while the D + C group exhibited the highest levels among the diabetic groups. The D + AT + HD and D + AT + MD groups had lower levels of ALT and AST enzymes compared to the other groups with no significant difference between D + AT + HD and D + AT + MD. D + AT + HD (p = 0.001), D + AT + MD (p = 0.001), D + HD (p = 0.023), D + MD (p = 0.029), and D + AT + oil (p = 0.011) upregulated LXRα compared to D + C. Among these groups, D + AT + HD exhibited a more profound upregulation of LXRα than D + AT + MD, D + AT + oil, D + HD, and D + MD (p = 0.005; p = 0.002, p = 0.001, and p = 0.001, respectively). Similarly, D + AT + HD showed a more notable upregulation of PGC-1α compared to D + AT + oil, D + HD, and D + MD (p = 0.002; p = 0.001, and p = 0.001, respectively). Pearson correlation tests showed significant and negative correlations between serum 25-hydroxyvitamin levels and both visceral fat (r = - 0.365; p = 0.005) and HOMA-IR (r = - 0.118; p = 0.009); while positive and significant correlations between the liver-to-bodyweight ratio with both ALT and AST enzymes and also between QUICKI levels with LXRα (r = 0.578; p = 0.001) and PGC-1α (r = 0.628; p = 0.001). Combined administration of aerobic training and vitamin D supplementation potentially improves liver enzymes in type-2 diabetic rats that were simultaneous with upregulating the levels of PGC-1α and LXRα proteins in hepatocytes. These improvements were more significant when combining exercise with high-dose vitamin D supplementation. This study highlights the potential of this combination therapy as a new diabetes treatment strategy.

Lung molecular and histological changes in type 2 diabetic rats and its improvement by high-intensity interval training.

BACKGROUND: Type 2 diabetes (T2D) leads to serious respiratory problems. This study investigated the effectiveness of high-intensity interval training (HIIT) on T2D-induced lung injuries at histopathological and molecular levels. METHODS: Forty-eight male Wistar rats were randomly allocated into control (CTL), Diabetes (Db), exercise (Ex), and Diabetes + exercise (Db + Ex) groups. T2D was induced by a high-fat diet plus (35 mg/kg) of streptozotocin (STZ) administration. Rats in Ex and Db + Ex performed HIIT for eight weeks. Tumor necrosis factor-alpha (TNFα), Interleukin 10 (IL-10), BAX, Bcl2, Lecithin, Sphingomyelin (SPM) and Surfactant protein D (SPD) levels were measured in the bronchoalveolar lavage fluid (BALF) and malondialdehyde (MDA) and total antioxidant capacity (TAC) levels were measured in lung tissue. Lung histopathological alterations were assessed by using H&E and trichrome mason staining. RESULTS: Diabetes was significantly associated with imbalance in pro/anti-inflammatory, pro/anti-apoptosis and redox systems, and reduced the SPD, lecithin sphingomyelin and alveolar number. Performing HIIT by diabetic animals increased Bcl2 (P < 0.05) and IL10 (P < 0.01) levels as well as surfactants components and TAC (P < 0.05) but decreased fasting blood glucose (P < 0.001), TNFα (P < 0.05), BAX (P < 0.05) and BAX/Bcl2 (P < 0.001) levels as well as MDA (P < 0.01) and MDA/TAC (P < 0.01) compared to the diabetic group. Furthermore, lung injury and fibrosis scores were increased by T2D and recovered in presence of HIIT. CONCLUSION: These findings suggested that the attenuating effect of HIIT on diabetic lung injury mediated by reducing blood sugar, inflammation, oxidative stress, and apoptosis as well as improving pulmonary surfactants components.

Chronic Electroacupuncture With High-Frequency at ST-36 Promotes Gastrointestinal Motility by Regulating Bone Morphogenetic Protein 2 Secretion of Muscularis Macrophages.

BACKGROUND: Electroacupuncture (EA) at Zusanli (ST36) is an alternative treatment for several gastrointestinal motility disorders; however, the exact mechanism is unconfirmed. We aimed to show the potential effects of EA on muscularis macrophages (MMφ), the bone morphogenetic protein (BMP)/BMP receptor (BMPR)-Smad signal pathway, and enteric neurons in diabetic mice. This may provide fresh insight into ways EA affects gastrointestinal motility. MATERIALS AND METHODS: C57BL/6J healthy adult male mice were randomly divided into five groups: regular control group, diabetes group, diabetes with sham EA group (acupuncture only), diabetes with low-frequency EA group (10 Hz), diabetes with high-frequency EA group (HEA) (100 Hz). The stimulation lasted eight weeks. Gastrointestinal motility was assessed. We identified M2-like MMφ in the layer of colonic muscle by flow cytometry. Western Blot, real-time polymerase chain reaction, and immunofluorescent staining were also used to determine the MMφ, molecules in the BMP2/BMPR-Smad pathway, and PGP9.5, neuronal nitric oxide synthase (nNOS) expression of enteric neurons in the colon of each group. RESULTS: 1) HEA improved the gastrointestinal motility (gastrointestinal transit time, defecation frequency) of diabetic mice. 2) HEA reversed the decreased proportion of M2-like MMφ and expression of the CD206 in the colon of diabetic mice. 3) HEA restored the downregulations of BMP2, BMPR1b, and Smad1 in the BMP2/BMPR-Smad pathway and increased downstream enteric neurons marked by PGP9.5, nNOS in the colon of diabetes mice. CONCLUSIONS: HEA might promote gut dynamics by upregulating M2-like MMφ in the colon of diabetic mice, which in turn leads to the accumulation of molecules in the BMP2/BMPR-Smad signaling pathway and downstream enteric neurons.

Bo's abdominal acupuncture improves disordered metabolism in obese type 2 diabetic rats through regulating fibroblast growth factor 21 and its related adipokines.

OBJECTIVE: To investigate the effect of Bo's abdominal acupuncture (BOAA) on fibroblast growth factor 21 (FGF21) and its related adipokines in type 2 diabetes mellitus (T2DM) rats. METHODS: This study established obese T2DM rat model by high-fat diet (HFD) with a dose of streptozotocin (STZ, 30 mg/kg). Obese T2DM rats were randomly subdivided into four groups (n = 10): negative, BOAA, conventional acupuncture (COA group) and metformin group (Met group) groups. The biochemical parameters, mRNAs, and proteins were analyzed using enzyme-lined immunoassays kits, quantitative polymerase chain reaction and Western blot. RESULTS: Treatment with BOAA attenuated the histopathological changes in visceral fat and restored the alterations in the levels of body weight, fasting blood glucose (FBG), homeostasis model assessment for insulin resistance (HOMA-IR). BOAA treatment significantly decreased the levels of triglyceride, total cholesterol, low density lipoprotein cholesterol, leptin, and increased the serum levels of high-density lipoprotein cholesterol, fibroblast growth factor 21 (FGF21), adiponectin (ADP), peroxisome proliferator-activated receptor γ (PPAR-γ), C-peptide (C-P) in obese T2DM rats. Furthermore, BOAA treatment significantly increased the mRNA expressions of FGF21, ADP, leptin, PPAR-γ, PPAR-α and adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK). Besides, BOAA treatment upregulated the protein expressions of fibroblast growth factor receptors3 (FGFR3), PPAR-α, extracellular signal-regulated kinase (ERK), phosphorylated ERK (p-ERK), AMPK, p-AMPK, Liver kinase B1 (LKB1), phosphorylated LKB1 (p-LKB1), acetyl-CoA carboxylase (ACC) and phosphorylated ACC (p-ACC), while downregulated the protein expressions of FGF21 and PPAR-γ in visceral fat. CONCLUSIONS: BOAA treatment reduced FBG and body weight, and improved insulin sensitivity through regulating FGF21 signaling pathway and its related adipokine in obese T2DM rats.

An Immunomodulatory Hydrogel by Hyperthermia-Assisted Self-Cascade Glucose Depletion and ROS Scavenging for Diabetic Foot Ulcer Wound Therapeutics.

Current therapeutic protocols for diabetic foot ulcers (DFUs), a severe and rapidly growing chronic complication in diabetic patients, remain nonspecific. Hyperglycemia-caused inflammation and excessive reactive oxygen species (ROS) are common obstacles encountered in DFU wound healing, often leading to impaired recovery. These two effects reinforce each other, forming an endless loop. However, adequate and inclusive methods are still lacking to target these two aspects and break the vicious cycle. This study proposes a novel approach for treating DFU wounds, utilizing an immunomodulatory hydrogel to achieve self-cascade glucose depletion and ROS scavenging to regulate the diabetic microenvironment. Specifically, AuPt@melanin-incorporated (GHM3) hydrogel dressing is developed to facilitate efficient hyperthermia-enhanced local glucose depletion and ROS scavenging. Mechanistically, in vitro/vivo experiments and RNA sequencing analysis demonstrate that GHM3 disrupts the ROS-inflammation cascade cycle and downregulates the ratio of M1/M2 macrophages, consequently improving the therapeutic outcomes for dorsal skin and DFU wounds in diabetic rats. In conclusion, this proposed approach offers a facile, safe, and highly efficient treatment modality for DFUs.

Compare the effectiveness of extracorporeal shockwave and hyperbaric oxygen therapy on enhancing wound healing in a streptozotocin-induced diabetic rodent model.

Studies have revealed that both extracorporeal shock-wave therapy (ESWT) and hyperbaric oxygen therapy (HBOT) can accelerate wound healing. This study aimed to compare the effectiveness of ESWT and HBOT in enhancing diabetic wound healing. A dorsal skin defect in a streptozotocin-induced diabetes rodent model was used. Postoperative wound healing was assessed once every 3 days. Histologic examination was performed with hematoxylin and eosin staining. Proliferation marker protein Ki-67 (Ki-67), endothelial nitric oxide synthase (eNOS), vascular endothelial growth factor (VEGF), and 8-hydroxy-2-deoxyguanosine (8-OHdG) were evaluated with immunohistochemical (IHC) staining. The wound area was significantly reduced in the ESWT and HBOT groups compared to that in the diabetic controls. However, the wound healing time was significantly increased in the HBOT group compared to the ESWT group. Histological findings showed a statistical increase in neovascularization and suppression of the inflammatory response by both HBOT and ESWT compared to the controls. IHC staining revealed a significant increase in Ki-67, VEGF, and eNOS but suppressed 8-OHdG expression in the ESWT group compared to the HBOT group. ESWT facilitated diabetic wound healing more effectively than HBOT by suppressing the inflammatory response and enhancing cellular proliferation and neovascularization and tissue regeneration.

Significant expansion of the donor pool achieved by utilizing islets of variable quality in the production of allogeneic "Neo-Islets", 3-D organoids of Mesenchymal Stromal and islet cells, a novel immune-isolating biotherapy for Type I Diabetes.

Novel biotherapies for Type 1 Diabetes that provide a significantly expanded donor pool and that deliver all islet hormones without requiring anti-rejection drugs are urgently needed. Scoring systems have improved islet allotransplantation outcomes, but their use may potentially result in the waste of valuable cells for novel therapies. To address these issues, we created "Neo-Islets" (NIs), islet-sized organoids, by co-culturing in ultralow adhesion flasks culture-expanded islet (ICs) and Mesenchymal Stromal Cells (MSCs) (x 24 hrs, 1:1 ratio). The MSCs exert powerful immune- and cyto-protective, anti-inflammatory, proangiogenic, and other beneficial actions in NIs. The robust in vitro expansion of all islet hormone-producing cells is coupled to their expected progressive de-differentiation mediated by serum-induced cell cycle entry and Epithelial-Mesenchymal Transition (EMT). Re-differentiation in vivo of the ICs and resumption of their physiological functions occurs by reversal of EMT and serum withdrawal-induced exit from the cell cycle. Accordingly, we reported that allogeneic, i.p.-administered NIs engraft in the omentum, increase Treg numbers and reestablish permanent normoglycemia in autoimmune diabetic NOD mice without immunosuppression. Our FDA-guided pilot study (INAD 012-0776) in insulin-dependent pet dogs showed similar responses, and both human- and canine-NIs established normoglycemia in STZ-diabetic NOD/SCID mice even though the utilized islets would be scored as unsuitable for transplantation. The present study further demonstrates that islet gene expression profiles (α, ß, γ, δ) in human "non-clinical grade" islets obtained from diverse, non-diabetic human and canine donors (n = 6 each) closely correlate with population doublings, and the in vivo re-differentiation of endocrine islet cells clearly corresponds with the reestablishment of euglycemia in diabetic mice. Conclusion: human-NIs created from diverse, "non-clinical grade" donors have the potential to greatly expand patient access to this curative therapy of T1DM, facilitated by the efficient in vitro expansion of ICs that can produce ~ 270 therapeutic NI doses per donor for 70 kg recipients.

"Adjust Zang and arouse spirit" electroacupuncture ameliorates cognitive impairment by reducing endoplasmic reticulum stress in db/db mice.

Introduction: Diabetic cognitive impairment (DCI) is a chronic complication of the central nervous system (CNS) caused by diabetes that affects learning and memory capacities over time. Recently, acupuncture has been shown to improve cognitive impairment in streptozotocin-induced diabetic rats. However, the effects of electroacupuncture on DCI and its underlying mechanism have not yet been elucidated in detail. Methods: In this study, we used db/db mice as DCI animal models which showed low cognitive, learning and memory functions. Electroacupuncture significantly ameliorated DCI, which is reflected by better spatial learning and memory function using behavioral tests. The db/db mice with cognitive impairment were randomly divided into a model group (Mod) and an electroacupuncture treatment group (Acup), while db/m mice were used as a normal control group (Con). First, the mice were subjected to behavioural tests using the Morris water maze (MWM), and body weight, blood glucose, insulin, triglycerides (TG) and total cholesterol (TC) were observed; HE, Nissl, and TUNEL staining were used to observe the morphological changes and neuronal apoptosis in the mice hippocampus; Finally, Western blot and rt-PCR were applied to detect the essential proteins and mRNA of ERS and insulin signalling pathway, as well as the expression levels of Tau and Aß. Results: Electroacupuncture significantly ameliorated DCI, which is reflected by better spatial learning and memory function using behavioral tests. Moreover, electroacupuncture attenuated diabetes-induced morphological structure change, neuronal apoptosis in the hippocampus of db/db mice. Our results revealed that electroacupuncture could regulate the expression levels of Tau and Aß by improving hippocampal ERS levels in db/db mice, inhibiting JNK activation, attenuating IRS1 serine phosphorylation, and restoring normal transduction of the insulin signaling pathway. Discussion: In summary, ERS and insulin signaling pathway paly causal roles in DCI development. Electroacupuncture can significantly alleviate the pathogenesis of DCI, improve mice's learning and memory ability, and improve cognitive dysfunction. This study adds to our understanding of the effect of acupuncture on DCI and opens the door to further research on DCI.

CTLA4-Ig alleviates the allogeneic immune responses against insulin-producing cells in a murine model of cell transplantation.

The adoptive transfer of insulin-producing cells (IPCs) is one of the promising treatments for insulin-dependent diabetes mellitus. While the use of allogeneic cell resources is inevitable in the case of a series of patients, alloimmune responses are a major barrier ahead of the successful implementation of allogeneic therapeutic cells. This study is aimed at evaluating the potential of CTLA4-Ig, as an approved immunomodulatory biologic, in protecting the IPCs against allogeneic immune responses. The C57BL/6 and BALB/c mice were used to establish a murine model of allogeneic cell transplantation. The mouse bone-marrow-derived mesenchymal stem cells were in vitro differentiated into IPCs, and the in vitro as well as the in vivo immune responses against IPCs were evaluated in the presence and absence of CTLA4-Ig. The allogeneic IPCs induced the in vitro activation of CD4+ T-cells, IFN-γ release, and the proliferation of lymphocytes, which all were controlled by CTLA4-Ig. Upon in vivo transfer of IPC into an allogeneic host, the splenic CD4+ and CD8+ T-cells exhibited a significant activation, and there was a significant donor-specific antibody response. Either of the mentioned cellular and humoral responses were modulated by a CTLA4-Ig regimen. This regimen also reduced the infiltration of CD3+ T-cells into the IPC injection site along with the improved overall survival of diabetic mice. CTLA4-Ig could be a complementary therapy for improving the efficacy of allogeneic IPC therapy through modulating the cellular and humoral responses that can lead to prolonged durability of IPCs within an allogeneic host.

Soy yoghurts produced with efficient GABA (γ-aminobutyric acid)-producing Lactiplantibacillus plantarum ameliorate hyperglycaemia and re-establish gut microbiota in streptozotocin (STZ)-induced diabetic mice.

Soy yogurt has been gaining popularity as a vegan food produced simply by soymilk fermentation with proper microbial manipulation. It is well known that soy containing rich isoflavones is beneficial for ameliorating hyperglycaemic disorders. Soy fermentation can improve the bioavailability of these precious nutrients. Lactiplantibacillus plantarum is one of the most abundant and frequently isolated species in soymilk manufacturing. Soy yogurts produced with efficient GABA (γ-aminobutyric acid)-producing L. plantarum and the deglycosylating activity of L. plantarum were functionally assessed in a STZ-induced hyperglycaemic mouse model. Hyperglycaemic mice were assigned into groups and treated with daily gavage of either dH2O, soymilk, soy yoghurts produced with high GABA-producing L. plantarum GA30 (LPGA30), low GABA-producing L. plantarum PV30 (LPPV30) or the soy yoghurts fortified with additional 30 mg g-1 GABA counterparts (GA + GABA and PV + GABA groups). Except the dH2O group, all soy yoghurt groups retained body weight with improved glucose homeostasis, glucose tolerance test results and renal tissue integrity, while the soymilk group shows partial benefits. Plasma GABA concentrations in the daily soy yoghurt-supplemented groups (LPGA30 and LPPV30) plateaued at 5 times higher than the average 0.5 µM in dH2O and soymilk groups, and their GABA-fortified soy yoghurt counterparts (GA + GABA and PV + GABA) groups were accountable for the restored plasma insulin levels. Gut microbiome analysis revealed dysbiosis in STZ-induced hyperglycemic mice of the dH2O group with breached out facultative anaerobic Proteobacteria over the normal phyla Firmicutes and Bacteroidetes. Restored gut microbiota with transitionally populated Actinobacteria was demonstrated in the LPGA30 group but not in the LPPV30 group. Soy yoghurts produced with efficient GABA-producing L. plantarum GA30 showed exceptional benefits in modulating gut microbiota with dominant genera of Enterococcus, Lactobacillus and Bifidobacterium, and the presence of some minor beneficial microbial communities including Akkermansia muciniphila, Butyricicoccus pullicaecorum, Corynebacterium spp. and Adlercreutzia spp. Efficient GABA-producing L. plantarum GA30 fermented soymilk to produce soy yoghurts that exhibit profound synergistic protections over rich soy isoflavones to restore pancreatic ß-cell functions for insulin production in STZ-induced hyperglycaemic mice. Additionally, the probiotic role of GABA-producing L. plantarum in re-establishing healthy gut microbiota in hyperglycaemic mice implies a possible symbiotic relationship, awaiting further exploration.

Interactive effect of swimming training and fenugreek (Trigonella foenum graecum L.) extract on glycemic indices and lipid profile in diabetic rats.

Present study investigated the interactive effect of swimming training and fenugreek on glycemic index and lipid profile of diabetic rats. Forty-eight diabetic rats were randomly assigned to (1) control(C), (2) training(T), (3) fenugreek(F), and (4) training + fenugreek(T + F) groups and 12 healthy rats were placed in healthy control (HC) group. During 4 weeks, groups 2 and 4 performed swimming training for 5 sessions per week and groups 3 and 4 received 100 mg/kg fenugreek. training, fenugreek and training + fenugreek significantly decreased glucose, insulin, insulin resistance, LDL, VLDL, TG and TC as well as increased HDL (p ≤ .05) also training + fenugreek had more favourable effects on improving glycemic indices and lipid profile compared to training and fenugreek alone (p ≤ .05). It seems that training and fenugreek alone or synergistically improve the glycemic indices and lipid profile in diabetic rats, nevertheless the synergistic effects of training and fenugreek can be more desirable than the effect of each alone.

Glucose metabolism-inspired catalytic patches for NIR-II phototherapy of diabetic wound infection.

Medical patches hold great prospects for diabetic wound administration, while their practical effects in diabetic wound management remain mired by the complexity of wound microenvironments. Here, inspired by the biological processes of glucose metabolism, we present a catalytic microneedle patch that encapsulates near-infrared-II responsive and dual-nanozyme active Au-Cu2MoS4 nanosheets (Au-CMS NSs) for treating diabetic wound infection. Since microneedle patches have great tissue penetration ability, the Au-CMS NSs can be delivered to deep tissues and fully interact with wound environments. Benefitting from the dual nanozyme activities (glucose oxidase and catalase) and near-infrared-II photothermal performances of Au-CMS NSs, the composited catalytic patch realizes in situ glucose consumption, oxygen generation, and bacterial elimination. Notably, their repeatability of near-infrared-II responsive antibacterial capability has been proved both in vitro and in diabetic mice against methicillin-resistant Staphylococcus aureus. The catalytic patch can find wide catalytic applications in wound care and infection prevention. STATEMENT OF SIGNIFICANCE: Effective treatment of diabetic wound infection remains still challenging in the clinic owing to the complex wound microenvironments. Herein, inspired by the biological processes of glucose metabolism in lives, we propose a novel strategy to treat wound infections by modulating the diabetic wound microenvironments. A near-infrared-II (NIR-II) responsive biocatalytic microneedle patch with both glucose oxidase- and catalase-like activities capable of killing bacteria, reducing glucose level, and supplying O2 is developed. The patch not only achieves efficient antibacterial outcomes in vitro, but also is a valuable wound patch for efficient treatment of MRSA-infected wounds in diabetic mice. We anticipate that this therapeutic strategy will provide the applications in chronic inflammation and infections.

[Electroacupuncture improves cognitive impairment of diabetic mice by regulating IRE1α-JNK pathway-related proteins in hippocampus].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on the cognitive impairment and expressions of inositol-dependent kinase 1α (IRE1α)/c-Jun N-terminal kinase (JNK) pathway-related proteins in diabetic mice, so as to explore its underlying mechanism. METHODS: Thirty db/db mice were randomly divided into model group (n=15) and EA group (n=15), and 15 db/m mice were chosen as the control group. EA was applied to "Baihui" (GV20) and "Shenting" (GV24), bilateral "Pishu" (BL20) and "Shenshu" (BL23), "Zusanli" (ST36) and "Sanyinjiao" (SP6) for 20 min, and bilateral "Feishu" (BL13), "Hegu" (LI4) and "Taichong" (LR3) were stimulated with filiform needles, with the needles retained for 20 min, once daily, 6 days a week for 4 weeks. The daily food intake, water intake, and weekly body weight and blood glucose of the mice in each group were recorded. The learning and memory abilities were detected by Morris water maze, the morphology of hippocampal cells was observed by HE staining, and IRE1α-JNK pathway-related proteins IRE1α, JNK, anti-apoptotic protein (Bcl-2) were detected by Western blot. RESULTS: Compared with the control group, the food intake, water intake, body weight, blood glucose in the mo-del group were significantly increased (P<0.01), the escape latency was significantly prolonged (P<0.05), the times of cros-sing platform were significantly reduced (P<0.01), and the expression levels of IRE1α and JNK proteins were significantly increased (P<0.01), while the expression of Bcl-2 protein was significantly decreased (P<0.01). Compared with the model group, the food and water intake in the EA group were significantly decreased (P<0.01), the body weight and blood glucose were significantly decreased (P<0.05, P<0.01), the escape latency was significantly shortened (P<0.05), the times of crossing platform significantly increased (P<0.05), and the expression levels of IRE1α and JNK proteins were significantly decreased (P<0.05), while the Bcl-2 expression was significantly increased (P<0.01). The cells in hippocampal CA1 area of mice in the model group are in disorder, with unclear nuclei and obvious vacuoles; while the morphology of nerve cells in EA group was significantly improved. CONCLUSION: EA can improve the cognitive impairment of db/db mice, as well as regulate body weight, blood glucose, and improve the cell morphology in the hippocampus, which may be related to its function in regulating the IRE1α-JNK pathway and related proteins.

Co-treatment with grape seed extract and mesenchymal stem cells in vivo regenerated beta cells of islets of Langerhans in pancreas of type I-induced diabetic rats.

BACKGROUND: Nowadays, diabetes mellitus is known as a silent killer because individual is not aware that he has the disease till the development of its complications. Many researchers have studied the use of stem cells in treatment of both types of diabetes. Mesenchymal stem cells (MSCs) hold a lot of potential for regenerative therapy. MSCs migrate and home at the damaged site, where they can aid in the repair of damaged tissues and restoring their function. Oxidative stress and inflammation represent a huge obstacle during MSCs transplantation. Therefore, the present study aimed to evaluate the role of grape seed extract (GSE) administration during MSCs transplantation in streptozotocin (STZ)-induced type I diabetes. Furthermore, testing some of GSE components [procyanidins(P)-B1 and P-C1] in conjunction with MSCs, in vivo, was performed to determine if one of them was more effective in relieving the measured attributes of diabetes more than the whole GSE. METHODS: Firstly, GSE was prepared from the seeds of Muscat of Alexandria grapes and characterized to identify its phytochemical components. Experimental design was composed of control group I, untreated diabetic group II, GSE (300 mg/kg)-treated diabetic group III, MSCs (2 × 106 cells/rat)-treated diabetic group IV and GSE (300 mg/kg)/MSCs (2 × 106 cells/rat)-treated diabetic group V. Type I diabetes was induced in rats by intravenous injection with 65 mg/kg of STZ. Treatment started when fasting blood glucose (FBG) level was more than 200 mg/dl; GSE oral administration started in the same day after MSCs intravenous injection and continued daily for 30 consecutive days. RESULTS: The results showed that GSE/MSCs therapy in type I-induced diabetic rats has dramatically managed homeostasis of glucose and insulin secretion; together with, improvement in levels of inflammatory markers and oxidative stress. CONCLUSION: Co-treatment with GSE and MSCs in vivo regenerates beta cells in type I-induced diabetic rats.

Electroacupuncture improves cognitive impairment in diabetic cognitive dysfunction rats by regulating the mitochondrial autophagy pathway.

BACKGROUND: Diabetes-associated cognitive dysfunction has become a major public health concern. However, the mechanisms driving this disease are elusive. Herein, we explored how electroacupuncture improves learning and memory function in diabetic rats. METHODS: The diabetic model was established by intraperitoneal injection of streptozotocin (STZ) in adult Sprague-Dawley rats. Rats were fed on high-fat and high-sugar diets. Learning and memory functions were assessed using behavioral tests. The hematoxylin and eosin (H&E) staining, Western blotting, real-time PCR, ELISA, immunohistochemistry, and transmission electronic microscopy (TEM) was performed to test related indicators. RESULTS: High-fat and high-sugar diets impaired learning and memory function in rats, while electroacupuncture treatment reversed these changes. The model group presented highly prolonged escape latency compared to the control group, indicating impaired learning and memory functions. The TEM examination showed that electroacupuncture enhanced Aß clearance and mitochondrial autophagy in hippocampal neuronal cells by increasing DISC1 expression. CONCLUSIONS: Electroacupuncture improves learning and memory function in diabetic rats by increasing DISC1 expression to promote mitophagy. This enhanced Aß clearance, alleviating cytotoxicity in hippocampal neuronal cells.

[Electroacupuncture preconditioning alleviates renal injury and oxidative stress through Sirt3/MnSOD pathway in type 2 diabetic rats].

OBJECTIVE: To explore the protective effect and molecular mechanism of electroacupuncture (EA) preconditioning on renal injury in type 2 diabetic rats. METHODS: Fifty male Wistar rats were randomly divided into control, model, EA, EA+inhibitor, and inhibitor groups, with 10 rats in each group. Diabetes model was established by high fat and high glucose diet and intraperitoneal injection of streptozotocin (40 mg/kg). EA (2 Hz, 1 mA) preconditioning was applied to "Guanyuan" (CV4), "Zhongwan" (CV12), bilateral "Zusanli" (ST36) and "Fenglong" (ST40) for 15 min, once every other day for 8 weeks. Rats of the inhibitor and EA+inhibitor groups were given intraperitoneal injection of 3-TYP (50 mg/kg) once every other day for a total of 3 times. The body weight, kidney mass, and renal index were recorded. The contents of urine microalbumin (ALB), 24 h urine 8-hydroxydeoxyguanosine (8-OHdG) and activities of reactive oxygen species (ROS), superoxide dismutase (SOD), catalase (CAT), glutathione glycine peroxidase (GSH-Px) in kidney were detected by ELISA. The activities of mitochondrial respiratory chain enzyme complex (RCCⅠ-RCCⅣ) in kidney were detected using spectrophotometric method. HE staining, Masson staining and transmission electron microscopy were used to observe the changes of renal structure. The protein and mRNA expressions of silent information regulator 3 (Sirt3), manganese superoxide dismutase (MnSOD) in kidney were detected by Western blot and quantitative real-time PCR, respectively. RESULTS: After modeling and compared with the control group, the contents of ALB, the renal index, activity of ROS and content of 8-OHdG, and the renal collagen volume fraction (CVF) were increased (P<0.01), while the activities of SOD, CAT and GSH-Px, RCCⅠ-RCCⅣ, and the mRNA and protein expressions of Sirt3 and MnSOD were decreased (P<0.01). After the treatment and compared with the model group, the contents of ALB, the renal index, ROS, 8-OHdG, and the CVF were decreased in the EA group(P<0.01, P<0.05), while the activities of SOD, CAT, GSH-Px, RCCⅠ-RCCⅣ, and Sirt3 and MnSOD expression levels were increased (P<0.01, P<0.05)；the RCCⅡ activity and the expression level of MnSOD mRNA were increased (P<0.05) in the EA+inhibitor group; the ALB and 8-OHdG contents and the CVF in the inhibitor group were increased (P<0.05), while the activity of SOD, and Sirt3 and MnSOD expression levels were decreased (P<0.05). In comparison with the EA group, the contents of ALB, the renal index, activities of ROS and 8-OHdG contents, and the CVF were increased (P<0.05, P<0.01), activities of SOD, CAT and GSH-Px and RCCⅠ and RCCⅡ, and the mRNA and protein expressions of Sirt3 and MnSOD were decreased (P<0.05, P<0.01) in both EA+inhibitor group and inhibitor group, whereas the activities of RCCⅢ and RCCⅣ were decreased in the inhibitor group (P<0.05). The therapeutic effect of inhibitor was notably inferior to that of EA+inhibitor in decreasing ALB and 8-OHdG contents, and CVF (P<0.01), and in up-regulating SOD and RCCⅡ activities, Sirt3 and MnSOD expression levels (P<0.01, P<0.05). CONCLUSION: EA preconditioning can increase the expressions of renal Sirt3 and MnSOD in type 2 diabetic rats, thereby reducing the oxidative stress response and protecting the kidneys.

[Effect of electroacupuncture at "Tianshu" (ST25) on the regulation of nitrergic neurons in different intestinal segments in streptozotocin induced type 2 diabetic rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) at "Tianshu" (ST25) on nitrergic neurons in jejunum and distal colon in type 2 diabetic rats, so as to explore its mechanism of regulating different intestinal segments. METHODS: Twenty-four SD rats were randomly divided into control, model and EA groups (n=8 in each group). The diabetes model was established by intraperitoneal injection of streptozotocin (35 mg/kg) and high-sugar and high-fat diet for 2 weeks. EA (2 Hz/15 Hz, 2 mA) was applied to bilateral ST25 for 20 min, once a day, 6 days a week for 4 weeks. The intestinal motility was evaluated by observing the first red stool excretion time and the distal colon bead excretion time. HE staining was used to observe the histological changes of jejunum and distal colon. The positive expression and protein expression of intestinal total neuronal marker protein gene product 9.5（PGP9.5） and neuronal nitric oxide synthase (nNOS) in jejunum and distal colon were detected by immunofluorescence staining and Western blot, respectively. RESULTS: After modeling, the blood glucose was significantly increased (P<0.01), the first red stool excretion time and the distal colon bead excretion time were shortened (P<0.01), the expression levels of PGP9.5 and nNOS in jejunum and distal colon were decreased (P<0.01) in the model group relevant to the control group. After treatment, compared with the model group, the blood glucose was decreased (P<0.01), the first red stool excretion time and the distal colon bead excretion time were prolonged (P<0.01, P<0.05), and the expression levels of PGP9.5 and nNOS in jejunum and distal colon were increased (P<0.05, P<0.01) in the EA group. HE staining showed disordered structure in intestinal mucosa of the jejunum and distal colon, and reduction of the number of goblet cells in the model group, which was relatively milder in the EA group. CONCLUSION: EA can effectively improve the intestinal mucosal damage and restore intestinal motor function in type 2 diabetic rats, which may be related to its function in regulating the number of nitrergic neurons in the intestinal nervous system.

Puffball spores improve wound healing in a diabetic rat model.

Persistent chronic oxidative stress is a primary pathogenic characteristics of diabetic foot ulcers. Puffball spores are a traditional Chinese medicine used to treat diabetic foot ulcers infections and bedsores. However, their effects against diabetic wounds and the mechanism underlying these effects remain largely unknown. The present study explored the effectiveness of puffball spores in diabetic wound treatment and the mechanisms underlying their effects. Sprague-Dawley rats with streptozotocin (STZ)-induced diabetes were treated with puffball spores to ascertain whether they accelerated wound healing.Real-time quantitative PCR, western blotting, hematoxylin-eosin and Masson's trichrome staining, immunohistochemistry analysis, and immunofluorescence assays were performed. As indicated by wound and serum histology and biochemical analyses, the puffball spores accelerated wound healing by activating Akt/Nrf2 signaling and promoting the expression of its downstream antioxidant genes, markedly stimulating antioxidant activity and enhanceing angiogenesis and collagen deposition. Our findings showed that puffball spores could accelerate diabetic wound healing, enhance antioxidant ability, promote the expression of vascular markers, and suppress inflammation, thus providing a theoretical basis for the treatment of diabetic and refractory wounds.

The effect of green coffee and/or endurance exercise on sperm function in pre-diabetic mice.

This study aimed to investigate the effect of green coffee (GC), chlorogenic acid (CA) as an active ingredient of GC and exercise, alone or in a combination with exercise, on spermatogenesis and sperm function in pre-diabetic mice. Results revealed that pre-diabetic status can have a significant adverse effect on spermatogenesis (Johnson score), and sperm concentration, motility, DNA damage and persistent histone in compared to the control group. Although lipid peroxidation, intracellular ROS production, and persistent histones in sperm were high in pre-diabetic mice, exercise only can improve sperm motility. GC alone only improved sperm motility in pre-diabetic mice while CA alone, even did not have this beneficial effect. However, GC along with exercise, did not improve motility but reduce DNA damage, while CA with exercise, significantly improved motility compared to pre-diabetic stage and to the level comparable to control. Therefore, based on this result in individuals with high DNA damage, GC supplementation and exercise could be useful approach while in asthenozoospermia, CA supplementation and exercise should be considered as an alternative approach. However, such an interpretation awaits validation.

[Electroacupuncture improves learning-memory ability by down-regulating expression of hippocampal Aß1-42 and Tau and NF-κB proteins in diabetic rats with cognitive impairment].

OBJECTIVE: To observe the effect of electroacupuncture(EA) on the expression of nuclear transcription factors κB (NF-κB) and Tau protein and content of amyloid (Aß) in diabetic rats with cognitive impairment, so as to explore its mechanism underlying improvement of learning-memory ability. METHODS: Male SD rats were randomly divided into normal control (n=9), model (n=9) and EA (n=9) groups. The diabetic mellitus model was established by feeding the rats with high fat and high sugar for 1 month and intraperitoneal injection of STZ (25 mg·kg-1·d-1) for 2 days. EA was applied to "Zusanli"(ST36) and "Neiting"(ST44) for 20 min, alternatively on both side every day, and "Yishu"(EX-B3) was stimulated by twirling the acupuncture needle with uniform reinforcing-reducing method for 1 min, followed by retaining it for 20 min. The treatment was conducted 6 times a week for 4 weeks. The learning-memory ability was evaluated by using Morris water maze swimming test. The blood glucose level was randomly detected by using a glucometer, the content of Aß1-42 in the hippocampal tissue was detected by ELISA, and the relative expression levels of hippocampal Tau and NF-κB p65 proteins and mRNAs were determined by Western blot and fluorescence quantitative real-time PCR, separately. RESULTS: After modeling, the blood glucose, escape latency, Aß1-42 content and the expression levels of Tau and NF-κB p65 proteins and mRNAs in the model group were significantly increased (P<0.01, P<0.05) in comparison with the normal control group. Following EA intervention, the modeling induced increase of blood glucose, escape latency, Aß1-42 content and the expression levels of Tau and NF-κB p65 proteins and mRNAs were reversed (P<0.05, P<0.01). CONCLUSION: EA can improve the learning-memory ability in rats with diabetic cognitive impairment, which may be related to its function in down-regulating the levels of hippocampal Aß1-42, Tau and NF-κB proteins.