SKF38393 prevents high glucose (HG)-induced endothelial dysfunction by inhibiting the effects of HG on cystathionine γ-lyase/hydrogen sulfide activity and via a RhoA/ROCK1 pathway.

BACKGROUND: Endothelial dysfunction plays a crucial role in diabetic vascular complications. A decrease in hydrogen sulfide (H2S) levels is increasingly becoming a vital factor contributing to high glucose (HG)-induced endothelial dysfunction. Dopamine D1-like receptors (DR1) activation has important physiological functions in the cardiovascular system. H2S decreases the dysfunction of vascular endothelial cells. However, no studies have reported whether DR1 protects the function of vascular endothelial cells by regulating H2S levels. AIM: The present study aimed to determine whether DR1 regulates the levels of endogenous H2S, which exerts protective effects against HG-induced injury of human umbilical vein endothelial cells (HUVECs) via Ras homolog gene family member A (RhoA)/Rho-associated coiled-coil containing kinase 1 (ROCK1) signalling. METHODS: HUVECs were exposed to HG (30 mM) or normal glucose (5.5 mM) after different treatments. Cell viability, proliferation and migration were measured by Cell Counting Kit-8, EdU cell proliferation assay, transwell assay and wound healing assay, respectively. H2S probe (7-Azido-4-Methylcoumarin) was used to detect levels of H2S. The intracellular calcium concentration ([Ca2+]i) were measured using Fluo-4 AM. The protein expressions were quantified by Western blot. RESULTS: We found that HG decreased the expression of DR1 and cystathionine γ-lyase (CSE) and H2S production. The DR1 agonist SKF38393 significantly increased DR1 and CSE expression and H2S production, whereas NaHS (a H2S donor) only increased CSE expression and H2S production but had no effect on DR1 expression. Meanwhile, SKF38393 further increased the [Ca2+]i induced by HG. In addition, HG reduced cell viability and the expression of Cyclin D1 and proliferating cell nuclear antigen and increased the expression of p21C⁢i⁢p/W⁢A⁢F-1, collagen I, collagen III, matrix metalloproteinase 9, osteopontin and α-smooth muscle actin and the activity of phosphorylated RhoA and ROCK1. SKF38393 and NaHS reversed these effects of HG. PPG (a CSE inhibitor) abolished the beneficial effect of SKF38393. These effects of SKF38393 were similar to those of Y-27632 (a ROCK inhibitor). CONCLUSION: Taken together, our results suggest that DR1 activation upregulates the CSE/H2S pathway by increasing the [Ca2+]i, which protects endothelial cells from HG-induced injury by inhibiting the RhoA/ROCK1 pathway.

[Effects of exogenous H2S on hepatic fibrosis in diabetic mice and its mechanism].

OBJECTIVE: To investigate the effects of exogenous hydrogen sulfide (H2S) on the hepatic fibrosis in diabetic mice and its mechanism. METHODS: Twenty-four C57 male mice (weight 22±2 g) were randomly divided into three groups (n=8): ① Normal control group (Control): Mice were intraperitoneally injected equal amount of normal saline, the injection time was the same as that of the experimental groups; ② Diabetes model groups (HG): Streptozotocin (STZ) was injected intraperitoneally once according to body weight (150 mg/kg) to establish diabetes model; ③ NaHS treatment groups (HG + NaHS): Mice were intraperitoneally injected with NaHS (100 µmol/L·kg·d) once a day for 12 consecutive weeks. The hepatocyte injury was detected by HE staining; the hepatic fibrosis was observed through Masson staining; the protein expressions of cystathionine - ß - synthetase (CBS), collagen-I (CoL-I), collagen-III (CoL-III) and matrix metalloproteinase-9 (MMP-9) were detected by Western blot. RESULTS: Compared with the control group, the damage and fibrosis of hepatocyte were significantly aggravated, the expression of CBS proteins was decreased (P＜0.01), and the expression levels of CoL-I, CoL-III and MMP-9 proteins were increased (P＜0.01) in the diabetic model group. Compared with the diabetic model group, the damage and fibrosis of hepatocyte were significantly lightened, the expression of CBS proteins was obviously increased (P＜0.01), and the expression levels of CoL-I, CoL-III and MMP-9 proteins were markedly decreased (P＜ 0.01). CONCLUSION: H2S inhibits the hepatic fibrosis in diabetic mice, and its mechanism is related to the decrease of collagen and matrix metalloproteinase-9.

Posterior percutaneous endoscopic versus traditional surgery for cervical disc herniation: A protocol for systematic review and meta analysis.

BACKGROUND: Posterior percutaneous endoscopic cervical diskectomy (P-PECD) can be used posterior microdiscectomy for cervical disc herniation. But only some small sample sizes of clinical studies have evaluated the efficacy and safety of P-PECD. This study aim to evaluated the efficacy and safety of P-PECD compared with traditional open surgery. METHODS: We will search the following seven electronic databases from their initiation to the May 1, 2020: PubMed, Embase, Cochrane Library, Web of Science, China National Knowledge Infrastructure (CNKI), Chinese Biomedical Literature Database (CBM) and Wanfang database. All randomized controlled trials, non-randomized controlled trials and retrospective case controls that compared the efficacy and safety of P-PECD and traditional open surgery in the treatment of cervical disc herniation will be included. The pooled odds ratio with 95% credible intervals (CIs) was used for the dichotomous variables. The mean difference with 95% CIs was used for the continuous variables. All analyses were conducted by Comprehensive Meta Analysis 2.0. A 2-tailed P value < 0.05 is considered statistically significant. RESULTS: The results of systematic review and meta-analysis will be submitted to a peer-reviewed journal. CONCLUSION: Our study will provide clarity regarding for clinicians to choices best surgical approach for patients with cervical disc herniation. Any changes that need to be made during the process of this study will be explained in the final full-text publication. PROTOCOL REGISTRATION NUMBER: CRD42020164011.

[Transforaminal percutaneous endoscopic lumbar discectomy combined with anchorage technique of pedicle for the treatment of high prolapse free lumbar disc herniation].

OBJECTIVE: To investigate the clinical effects of percutaneous spinal endoscopy (percutaneous endoscopy) in the treatment of high prolapse free lumbar disc herniation. METHODS: From May 2016 to June 2018, 24 patients with highly prolapse free lumbar disc herniation were enrolled in this study, including 11 males and 13 females, ranging in age from 48 to 72 years old, with an average of (59.5±7.2) years old. There were 1 case of L2, 3, 5 cases of L3, 4, 18 cases of L4, 5. The course of disease ranged from 8 to 26 months, with an average of (16.2±6.3) months. All the patients were subjected to local infiltration anesthesia. The clinical outcomes were evaluated by visual analog scale (VAS) for leg pain, low backpain and Oswestry Disability Index (ODI) at preoperative, first day after operation and 6 month after surgery. All data were statisticed by SPSS 22.0. RESULTS: All the patients were followed up, and the duration ranged from 12 to 24 months, with a mean of (17.5±5.3) months. The average operation time was(69.8±14.2) minutes. One patient had cerebrospinal fluid leakage, which improved after supine rest. VAS scores of lower back pain were 6.36±1.27, 3.94±1.03, 1.62±0.87, 0.44±0.27, 0.37±0.29. VAS scores of leg pain were 8.28±1.74, 3.16±1.24, 2.83±1.13, 0.83±0.31, 0.46±0.31, and the differences were statistically significant (P<0.05). The ODI were (48.79±9.83)%, (36.51±11.24)%, (21.05±6.35)%, (9.83±4.62)% and (7.24±4.72)% 1 day before and 1 weeks, 3 months, 6 months and 1 year after the operation, respectively. One year after the operation, the modified Macnab evaluation system was used to evaluate the clinical efficacy of the patients, 19 patients got an excellent result, 3 good, 2 fair and 0 poor. CONCLUSION: Percutaneous endoscopic pedicle anchoring technique for the treatment of high prolapse free lumbar disc herniation can effectively improve the clinical symptoms of patients, and has the advantages of less trauma, less bleeding, rapid recovery, complete removal of the nucleus pulposus, and less pain of patients, etc., its clinical efficacy is accurate, operability is strong, it is worth promoting in the clinical use.