Effects of Compound Centella on Oxidative Stress and Keap1-Nrf2-ARE Pathway Expression in Diabetic Kidney Disease Rats.

The formula of Compound Centella mainly contains 3 traditional Chinese herbs: Centella asiatica (L.) Urb. (JiXueCao), Astragalus Membranaceus Fish. (HuangQi), and Tripterygium wilfordii Hook. f. (LeiGongTeng). Though this formula is effective for treating diabetic kidney disease (DKD) in clinic, the exact mechanism is still unclear. This study aims to investigate the effect and antioxidant mechanism of Compound Centella on DKD rats. High-performance liquid chromatography (HPLC) was applied to analyse 3 herbs in Compound Centella. Sprague-Dawley rats were divided into the normal group (NG), DKD group (DKDG), Compound Centella group (CCG), and losartan group (LG), with 8 rats in each group. On the first day of the experiment, rats in the NG were fed with ordinary -feed, while the other groups were fed with high-fat and high-sugar feed. On the 29th day, except the NG, the other 3 groups received a single intraperitoneal injection of streptozocin (STZ, 35 mg/kg). Fasting blood glucose (FBG) was measured on the 1st day, 32nd day, 46th day, 56th day, 84th day, and 112th day. Total protein/creatinine ratio of urine was determined by the phenol red assay on the 1st day and 112th day. Serum creatinine (Scr) was determined by an automatic biochemical analyser on the 112th day. Kidneys were collected on the 112th day for analysis and evaluation. Periodic acid-Schiff (PAS) staining, hematoxylin-eosin (HE) staining, and transmission electron microscopy were used to observe kidney pathological changes. The mRNA and protein expressions of Kelch-like ECH-associated protein 1 (Keap1) and nuclear factor-erythroid 2-related factor 2 (Nrf2) in renal tissues were detected by RT-qPCR, Western blot, and immunohistochemistry. Oxidative stress was evaluated by detecting the levels of malondialdehyde (MDA) and heme oxidase-1 (HO-1). The results showed that the content of asiaticoside, astragaloside, and triptolide in the herb was 5960, 809, and 2.42 µg/g and in the Compound Centella was 340, 64, and 0.1403 µg/mL by HPLC. Compound Centella reduced the urine protein/creatinine ratio and improved renal pathology in the kidneys of DKD rats. In addition, the mRNA and protein expressions of Keap1 and Nrf2 in kidneys were upregulated by Compound Centella. The expressions of MDA were downregulated but HO-1 were upregulated by Compound Centella. Therefore, the protective effect of Compound Centella in the kidney of DKD rats may be related to regulating the Keap1-Nrf2-ARE pathway under oxidative stress, suggesting Compound Centella as a promising treatment against DKD.

Jixuecao (Herba Centellae Asiaticae) alleviates mesangial cell proliferation in IgA nephropathy by inducing mitofusin 2 expression.

OBJECTIVE: To investigate the effect of mitofusin 2 (Mfn2) and its downstream signaling pathway on glomerular mesangial cells (GMCs) proliferation in IgA nephropathy (IgAN), as well as the mechanism of action of Jixuecao (Herba Centellae Asiaticae, HCA) in the treatment of IgAN. METHODS: Adenovirus-mediated Mfn2 gene transfection and Mfn2 expression were analyzed by real-time polymerase chain reaction (PCR) and Western blotting. IgA1 induced the proliferation of GMCs, which were then treated with HCA. Cell proliferation was detected with cell counting kit-8 (CCK-8), and Mfn2 expression was analyzed by real-time PCR and western blotting. An IgAN animal model was also established and treated with HCA. GMCs proliferation was detected by hematoxylin-eosin staining, mitochondrial structure was analyzed by electron microscopy, mitochondrial function was determined by the Clark oxygen electrode method, and the expression of Mfn2, Phospho-extracellular regulated protein kinases1/2 (P-ERK1/2), Cyclin-dependent kinase 2 (CDK2), Phospho-p27 (p-p27), and cyclin A was analyzed by Western blotting. RESULTS: In vitro, HCA inhibited GMCs in a concentration-dependent manner in association with the upregulation of Mfn2 expression. The overexpression of Mfn2 inhibited IgA1-induced GMCs proliferation and elevated the effect of HCA. In vivo, treatment with HCA could alleviate albuminuria and creatinine and GMCs proliferation. These effects were related to the upregulation of Mfn2, p-p27 and inhibition of p-ERK1/2, CDK2, and cyclinA. Mitochondrial swelling, vacuolar degeneration, and reduction of respiratory control rate were identified in IgAN, but HCA could improve the mitochondrial structure and function. CONCLUSION: HCA inhibited GMCs proliferation via the upregulation Mfn2 and the inhibition of Ras-Raf-ERK/MAPK. We revealed that changes of mitochondrial structure and function are associated with IgAN, but that HCA can improve these mitochondrial features.

Downregulation of microRNA-448 improves isoflurane-induced learning and memory impairment in rats.

The present study aimed to investigate the potential role of microRNA­448 (miR­448) in isoflurane-induced learning and memory impairment in rats. Sprague­Dawley rats were used for the construction of isoflurane­treated models. The Morris water maze test was used to evaluate the effects of isoflurane on rats regarding the following para-meters: Swimming speed, escape latency and time in original quadrant. Influences of isoflurane on neuron apoptosis and miR­448 expression in rat hippocampus tissue were analyzed by flow cytometry and reverse transcription­quantitative polymerase chain reaction, respectively. Furthermore, the effects of miR­448 on the expression of cell apoptosis­associated proteins were investigated by flow cytometry. The results demonstrated that isoflurane treatment induced higher escape latency and lower time spent in original quadrant compared with the control rats. In addition, isoflurane treatment induced neuron apoptosis and miR­448 was highly expressed in the hippocampal tissue of isoflurane­treated rats. Furthermore, Bcl­x was significantly downregulated while caspase­3 expression was upregulated by an miR­448 inhibitor. Combined the results of the current study indicate that miR­448 knockdown may have pivotal roles in improving isoflurane-induced learning and memory impairment via suppressing neuron apoptosis.

COX-2 inhibition attenuates lung injury induced by skeletal muscle ischemia reperfusion in rats.

BACKGROUND: Skeletal muscle ischemia reperfusion accounts for high morbidity and mortality, and cyclooxygenase (COX)-2 is implicated in causing muscle damage. Downregulation of aquaporin-1 (AQP-1) transmembrane protein is implicated in skeletal muscle ischemia reperfusion induced remote lung injury. The expression of COX-2 in lung tissue and the effect of COX-2 inhibition on AQP-1 expression and lung injury during skeletal muscle ischemia reperfusion are not known. We investigated the role of COX-2 in lung injury induced by skeletal muscle ischemia reperfusion in rats and evaluated the effects of NS-398, a specific COX-2 inhibitor. METHODS: Twenty-four Sprague Dawley rats were randomized into 4 groups: sham group (SM group), sham+NS-398 group (SN group), ischemia reperfusion group (IR group) and ischemia reperfusion+NS-398 group (IN group). Rats in the IR and IN groups were subjected to 3h of bilateral ischemia followed by 6h of reperfusion in hindlimbs, and intravenous NS-398 8 mg/kg was administered in the IN group. In the SM and SN groups, rubber bands were in place without inflation. At the end of reperfusion, myeloperoxidase (MPO) activity, COX-2 and AQP-1 protein expression in lung tissue, PGE2 metabolite (PGEM), tumor necrosis factor (TNF)-α and interleukin (IL)-1ß levels in bronchoalveolar lavage (BAL) fluid were assessed. Histological changes in lung and muscle tissues and wet/dry (W/D) ratio were also evaluated. RESULTS: MPO activity, COX-2 expression, W/D ratio in lung tissue, and PGEM, TNF-α and IL-1ß levels in BAL fluid were significantly increased, while AQP-1 protein expression downregulated in the IR group as compared to that in the SM group (P<0.05). These changes were remarkably mitigated in the IN group (P<0.05). NS-398 treatment also alleviated histological signs of lung and skeletal muscle injury. CONCLUSION: COX-2 protein expression was upregulated in lung tissue in response to skeletal muscle ischemia reperfusion. COX-2 inhibition may modulate pulmonary AQP-1 expression and attenuate lung injury.

Fentanyl inhibits cell viability in human pancreatic cancer cell line and tumor growth in pancreatic cancer cell-transplanted mice.

Pancreatic cancer is a kind of devastating disease with a high mortality rate. Fentanyl has been widely applied to anesthesia and analgesia in pancreatic cancer therapy, and is also demonstrated to inhibit the growth of some kinds of cancer cells in existed studies. To investigate the functions of fentanyl in pancreatic cancer, we conducted a series of in vivo and in vitro experiments using human pancreatic cancer cells SW1990 and fentanyl treatment. The cells were transplanted to BALB/c nude mice to generate pancreatic tumor for monitoring tumor growth. Viability, apoptosis, migration and invasion, and cell cycle of SW1990 cells were also analyzed. To reveal the functional mechanisms of fentanyl, the expression changes of factors in these cellular activities were detected. Results showed a significant inhibition of pancreatic tumor growth in the fentanyl-treated group. Fentanyl also inhibited viability of SW1990 cells in vitro. Detailed results showed fentanyl led to promoted cell apoptosis via arresting cells in G0/G1 phase. It also suppressed cell migration and invasion. Further proofs indicated that the factors related to cell apoptosis (Bcl-2, p53 and Caspase-3), cell cycle (p21, Cyclin D1 and CDK4) and epithelial-mesenchymal transition (E-cadherin, Vimentin and α-SMA) showed the corresponding expression changes. Fentanyl might execute its functions via the suppressed MAPK pathways, since the key factors, p38, ERK1/2 and JNK were all down-regulated by fentanyl. This study indicated fentanyl could inhibit viability and growth of pancreatic cancer cells, providing a possible strategy for pancreatic cancer treatment.

The analgesic efficacy of subcostal transversus abdominis plane block compared with thoracic epidural analgesia and intravenous opioid analgesia after radical gastrectomy.

BACKGROUND: The transversus abdominis plane (TAP) block has been shown to provide effective postoperative analgesia in lower abdominal surgery. Subcostal TAP block has also been proposed as a new technique to provide analgesia for the supraumbilical abdomen. We compared the analgesic and opioid-sparing effects of a single-injection subcostal TAP block with continuous thoracic epidural analgesia and IV opioid analgesia. METHODS: Ninety patients undergoing elective radical gastrectomy were randomized to receive either combined general-subcostal TAP anesthesia (group TAP), combined general-epidural anesthesia (group EA), or general anesthesia (group GA), and were analyzed on an intention-to-treat basis. In group TAP, a bilateral subcostal TAP block was performed after induction of general anesthesia using 20 mL of 0.375% ropivacaine. In group EA, a thoracic epidural was placed between T8 and T9 and bolused with 8 mL of 0.25% ropivacaine before induction of general anesthesia. The epidural was maintained with 5 mL/h of 0.25% ropivacaine during the surgery. Group GA received standard general anesthesia. In the postanesthesia care unit (PACU), all groups received IV morphine titration for visual analog scale (VAS) pain scores >3. All patients were started on IV patient-controlled analgesia with morphine after morphine titration in the PACU, while group EA also had their epidural maintained with 5 mL/h of 0.125% bupivacaine with 8 µg/mL morphine. Patients were assessed in the PACU and at 1, 3, 6, 24, 48, and 72 hours postoperatively. Primary outcomes measured were morphine consumption at 24 hours and all VAS pain scores. RESULTS: Data from 82 of 90 (91.1%) patients were included in the study. Group TAP demonstrated decreased cumulative morphine consumption at 24 hours (98.75% confidence intervals, -29 to -9 mg) and noninferiority on VAS pain scores at all measurement times, as compared with group GA with standard opioid analgesia. However, group EA was superior to group TAP regarding cumulative morphine consumption at 24 hours (98.75% confidence intervals, -23 to -4 mg) and noninferior to group TAP on VAS pain scores at all comparison points. Group TAP had reduced morphine consumption from PACU admission to 6 hours as compared with group GA, but increased morphine consumption for 6 to 24 hours as compared with group EA. CONCLUSION: Single-injection subcostal TAP block was more effective than IV opioid analgesia, while continuous thoracic epidural analgesia was more effective than the single-injection subcostal TAP block.

[Protective effect of Shenmai injection on lung injury induced by cardiac pulmonary bypass].

OBJECTIVE: To observe the protective effect of Shenmai Injection (SMI) on acute lung injury (ALI) induced by cardiac pulmonary bypass (CPB). METHODS: Thirty patients, scheduled to receive cardiac valve replacement by CPB for the first time, were equally randomized into 2 groups, the treated group and the control group. Cardiac valve replacement was performed under extracorporeal circulation after general anaesthesia. SMI 0.6 mL/kg was given to the treated group by adding in 250 mL physiological saline for intravenous dripping at the time between intubation under anaesthesia and CPB, while 250 mL physiological saline was given to the control group alone. Blood-gas analysis was performed with blood withdrawal from the radial artery to record PaO2, PaCO2, fraction of inspired oxygen (FiO2), by them the alveolar-arterial difference of partial oxygen pressure [P(A-a) DO2] was calculated, and the respiratory index (RI), the blood concentrations of soluble intercellular adhesion molelue-1 (sICAM-1), endothelin-1 (ET-1) and nitric oxide (NO) were measured at various time points, i.e. before anesthesia induction, 0.5 h, 2 h, 6 h and 24 h after ending CPB. RESULTS: All indices wer not significantly different between the two groups before operation (P > 0.05). After CPB, P(A-a) DO2 and RI were gradually elevated and reached the peak at 2 h after ending CPB, the increment in the treated group was lower than that in the control group (P < 0.05 or P < 0.01). Compared with the P(A-a) DO2 before anesthesia induction, P (A-a) DO2 at 0.5-24 h was statistically different (P < 0.05 or P < 0.01). Compared with the RI before anesthesia induction, RI at 2-24 h was statistically different (P < 0.05 or P < 0.01). Blood concentrations of sICAM-1 gradually raised after CPB, and reached the peak at 2 h after ending CPB, showed a higher level at 0.5-6 h after ending CPB as compared with that before anesthesia induction (P < 0.05 or P < 0.01). Blood ET-1 showed a figure of increasing as sICAM did (P < 0.05 or P < 0.01), and with a lower degree at 0.5-2 h after ending CPB in the treated group (P < 0. 01). Blood NO obviously reduced after CPB, it was lower at 0.5-24 h after ending CPB than at that before anesthesia induction (P < 0.01), and the decrement was lesser in the treated group (P < 0.05 or P < 0.01). CONCLUSION: SMI can attenuate the acute lung injury after CPB by way of inhibiting vascular endothelial cell adhesion with inflammatory cells, antagonizing lipid peroxidation, and improving the ventilation and oxygenation function of lung.

[Effect and significance of Shenmai injection on value of vascular endothelial active factors of heart valve replacement patients].

OBJECTIVE: To observe the effect of Shenmai injection on the value of vascular endothelial active factors nitric oxide (NO), endothelin-1 (ET-1) of patients undergoing heart valve rveplacement and cardiac pulmonary bypass (CPB). METHOD: Thirty patients of cardiac valve replacement undergoing open heart surgery under cardiopulmonary bypass (CPB) were randomized single-blind method divided into Shenmai injection group (SM) and control group (C) with 15 cases each. Shenmai injection group (SM) were injected Shenmai injection 0.6 mL x kg(-1) added to physiological saline 250 mL after anaesthesia before CPB, the control control group were injected only physiological saline 250 mL at the same time. Blood samples were taken before induction of anesthesia and at 0.5, 2, 6, 24 hours after terminating CPB. To calculate P(A-a) DO2 and respiratory indexs (RI) by blood gas analysis, nitric oxide (NO) and endothelin-1 (ET-1) ET-1 were measured. At the same time, the time of CPB and artery blockage were recorded. RESULT: There was no statistical significance before operation between 2 groups to every blood index. After CPB, P(A-a) DO2 , RI and ET-1 was higher than pre-operation after CPB (P < 0.05). But P(A-a) DO2, RI and ET-I of Shenmai group were lower than control group evidently at every point after CPB (P < 0.05 or P < 0.01). The concentrations of NO were reduced obveiously after CPB (P < 0.05), but in shenmai group, the range of descent was lower than control group (P < 0.01 or P < 0.05). CONCLUSION: The concentrations of NO and ET-1 is connected with the lung injury after CPB. Through rise the level of NO and reduce the level of ET-1, Shenmai injection can alleviate the lung injury in some degree after CPB and improve pulmonary oxygenation function.