ABSTRACT
Objective:To observe the effect of Shaofu Zhuyutang on nuclear factor erythroid-2-related factor 2 （Nrf2） /antioxidant response element （ARE） signaling pathway in blood vessels by establishing the model of rats with cold coagulation and blood stasis syndrome， and to explore the protective effect and mechanism of Shaofu Zhuyutang on vascular endothelial injury. Method:The 50 SPF rats were randomly divided into high dose group （4.8 g·kg-1）， middle dose group （2.4 g·kg-1）， low dose group （1.2 g·kg-1）， model group and normal group （ten of each group）. The rat model of cold coagulation and blood stasis syndrome was established by subcutaneous injection of epinephrine hydrochloride combined with ice bath. At the same time of modeling， the drug was administered by gavage. After 28 days of continuous administration， the hemorheology indexes were detected by automatic hemorheology instrument. Levels of nitric oxide （NO）， endothelin （ET）-1， superoxide dismutase （SOD）， glutathione （GSH-Px）， intercellular adhesion molecule-1（ICAM-1）， vascular cell adhesion molecule-1（VCAM-1）， von Willebrand factor （vWF） in serum were determined by ELISA. Hematoxylin and eosin （HE） staining was used to observe the endothelial injury of vascular tissue of thoracic aorta. The protein expression of Nrf2 and HO-1 in vascular tissue of thoracic aorta was detected by Western blot. Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）was used to observe the expression of Nrf2 and heme oxygenase-1 （HO-1） mRNA in vascular tissue of thoracic aorta. Result:Compared with the blank group， model group rats whole blood viscosity and plasma viscosity were significantly increased （P<0.05，P<0.01）， vWF， ICAM 1， VCAM 1 content increased significantly （P<0.01）， NO， SOD， gsh-px levels decreased significantly （P<0.01）， significantly increased the content of ET-1（P<0.01）， thoracic aorta vascular tissue Nrf2， HO-1 mRNA expression was significantly increased （P<0.01）， Nrf2 protein expression in the cell nucleus increased significantly （P<0.05）， The protein expression level of Nrf2 in cytoplasm was significantly decreased （P<0.05）， while the protein expression level of HO-1 was significantly increased （P<0.01）. Compared with model group， the whole blood viscosity （high and middle cut）， plasma viscosity， were significantly reduced in high and meduim-dose Shaofu Zhuyutang groups（P<0.05，P<0.01）. The levels of vWF， ICAM-1， VCAM-1 and ET-1 in serum were significantly reduced （P<0.05，P<0.01）， NO， SOD and GSH-Px increased significantly （P<0.05，P<0.01）. The pathological changes such as hyperplasia， swelling and shedding of endothelial cells of thoracic aorta， rupture of internal elastic membrane and disorder of smooth muscle arrangement were improved. The expression levels of Nrf2， HO-1 protein and gene were significantly increased in vascular tissue of thoracic aorta （P<0.01）. Conclusion:Shaofu Zhuyutang has a protective effect on vascular endothelial injury in rats with cold coagulation and blood stasis syndrome. The mechanism of action is related to the activation of Nrf2/ARE signaling pathway， which leading to the increased expression of antioxidant enzymes and decreased the expression of adhesion factors.
ABSTRACT
BACKGROUND: Caspofungin, a novel echinocandins systemic antifungal agent, has been shown to exert broad-spectrum antibacterial effect on deep fungal infections, which is superior to or equivalent with the role of amphotericin B, but there is no report on its application for preventing fungal infection after renal transplantation.OBJECTIVE: To analyze the difference in high risk factors of fungal infection after kidney transplantation using donation after cardiac death donors and living-related donor kidney transplantations, and to explore the feasibility and safety of caspofungin to prevent fungal infection after kidney transplantation using donation after cardiac death donors.METHODS: This was a prospective, single-center, controlled trial finished at the Department of Kidney Transplantation,the First Affiliated Hospital of Zhengzhou University, Henan Province, China. Totally 188 patients undergoing primary kidney transplantation without history of fungal infection and use of antifungal drugs between January 2012 and August 2013 were enrolled, including kidney transplantation with donation after cardiac death donors (n=102, trail group), and kidney transplantation with living-related donors (n=86, control group). The CYP3A5 genotype was determined preoperatively. All patients received tacrolimus+mycophenolate mofetil+prednisone triple immunosuppression after transplantation. The trial group was subjected to caspofungin therapy for 2 weeks. The risk factors for fungal infection in the two groups were compared, and the effects of caspofungin on the tacrolimus concentration, tacrolimus concentration/dose were detected in the recipients with same CYP3A5 genotype recipients at 1 and 2 weeks, and 1, 3 and 6 months postoperatively. The liver and kidney function, adverse events and fungal infections were recorded at different time points. This trial was registered with the Chinese Clinial Trial Registry (Regitration number:ChiCTR-OON-17013342).RESULTS AND CONCLUSION: The survival rate of patient/kidney was 98.4% and 97.3% respectively, 97 cases in the trial group and 86 controls competed 6-month follow-up. Preoperative hemodialysis time, hemoglobin value, cold ischemia time, warm ischemia time, intraoperative blood transfusion volume, time of central venous catheter kept in situ,methylprednisolone usage, ATG usage, serum creatinine reduced level at 1 week, thrombocytopenia and duration of postoperative body temperature > 38 ℃ were the risk factors for fungal infection in the trail group relative to the control group. The fungal infection rate in the trial and control groups was 0% and 2.3%, respectively, at 6 months of follow-up.The serum creatinine level in the trail group was significantly higher than that in the control group at 1 month postoperatively (P < 0.05), and the level showed no significant difference between two groups at other time points (P >0.05). After 2 weeks of caspofungin treatment, the concentrations of tacrolimus and tacrolimus concentration/dosage did not differ significantly in different CYP3A5 genotype recipients (P > 0.05). Caspofungin might induce some adverse reactions, especially electrolyte disturbance with an incidence of 21.6%, but there was no significant difference between two groups (P > 0.05). These findings imply that kidney transplantation using donation after cardiac death donors presents with various risk factors for fungal infection compared with living-related donor kidney transplantation.Furthermore, caspofungin is effective and safe for preventing fungal infection and has no effect on tacrolimus concentration; therefore, it can be used as a new anti-fungal agent after kidney transplantation.