3.Protective effects of notoginsenoside R1 against amyloid-β (1-42) induced mitochondrial apopototic death in SH-SY5Y cells.
Tao MA ; Wen-feng XIN ; Wen-sheng ZHANG ; Yong-yan WANG
China Journal of Chinese Materia Medica 2015;40(2):303-307
OBJECTIVETo investigate the effects and underlying mechanism of notoginsenoside R1 on amyloid-β (1-42) (Aβ(1-42)) induced mitochondrial apoptotic death in SH-SY5Y cells.
METHODCell viability was assayed by MTT, apoptotic rates were analyzed with PI/Annexin V flow cytometry, Bax and Bcl-2 expression were detected with Western blotting, enzymatic activity of caspase-3, caspase-8 and caspase-9 were measured by ELISA assay.
RESULTThe 6.25-100 nmol x L(-1) of notoginsenoside R1 attenuate Aβ(1-42) induced apoptotic death of SH-SY5Y in dose dependent manner. The ratio of Bcl-2/Bax was elevated in SH-SY5Y with notoginsenoside R1 treatment. Caspase-3 and caspase-9 were activated with notoginsenoside R1 treatment while caspase-8 was not affected.
CONCLUSIONNotoginsenoside R1 could protect SH-SY5Y cells from Aβ(1-42) induced apoptosis via mitochondria related apoptotic pathway.
Amyloid beta-Peptides ; antagonists & inhibitors ; Apoptosis ; drug effects ; Caspases ; metabolism ; Cell Line, Tumor ; Cell Survival ; drug effects ; Cytoprotection ; Ginsenosides ; pharmacology ; Humans ; Mitochondria ; drug effects ; Peptide Fragments ; antagonists & inhibitors
4.Resveratrol protects against oxidative damage of fibroblasts irradiated with UVB
Yuehua YAN ; Xin ZHOU ; Tao SONG ; Jianbo WU ; Wanxiang SHENG ; Xiaoming LIU ; Chunrong YUAN
Chinese Journal of Physical Medicine and Rehabilitation 2010;32(7):503-505
Objective To explore the protective effect of resveratrol against oxidative damage to cultivated fibroblasts irradiated with UVB. Methods Fibroblasts from normal human skin cultured in vitro were divided into 5 groups (a normal control group, a group irradiated with UVB, a group treated with resveratrol before UVB irradiation, and a group treated after irradiation). A monolayer of fibroblasts was irradiated with UVB at 60 mJ/cm2. The vitality of the cells was measured using the methylthiazol tetrazolium (MTT) method. The activity of superoxide dis-mutase (SOD) and malondialdehyde (MDA) content were determined using enzyme biochemistry. Results Resveratrol over 100 μM inhibited the proliferation of fibroblasts. Resveratrol under 100 μM improved the proliferation of cells. The optimal concentration was 50 μM. UVB irradiation decreased the vitality of the cells and SOD activity, and it significantly enhanced MDA content. Conclusions Resveratrol treatment before or after UVB irradiation elevates the survival rate of fibroblasts, enhances the activity of SOD, and decreases MDA content. Resveratrol at low concentration could improve the proliferation of fibroblasts, and at high concentration could inhibit their proliferation. Res-veratol at 50 μM relieves the inhibited proliferation of fibroblasts damaged by UVB irradiation.
5.The experimental study on infant rabbit lung injury induced by ischemia-reperfusion
Wanshan QIU ; Bing JIA ; Ming YE ; Xiangang YAN ; Gang CHEN ; Qilin TAO ; Sheng SHEN ; Zhanggen CHEN
Chinese Journal of Thoracic and Cardiovascular Surgery 2012;(12):729-731
Objective To explore the characteristics of ischemia-reperfusion induced infant lung damage and the potential mechanisms of the injuried.Methods Both infant (15-21 days old) and adult (5-6 months old) rabbits were subjected to either ischemia-reperfusion or sham operation.Ischemia-reperfusion was induced by clamping the right pulmonary hilum for 1 hour and then removal of the clamp for 4 hours under anesthesia.The lung tissue were sampled for histological examination by light and electron microcopies and for biological evaluation of mitochondrial alterations.Production and expression of free radical species-hydroxyl radical (ROS-HR),malondialdehyde (MDA),superoxide dismutase (SOD),glutathione peroxidase (GSH-PX),myeloid differentiation factor-88 (MyD-88),and nuclear factor-κB (NF-κB) in the lung tissue were also examined.In addition,circulating levels of interleukin-β and tumor necrosis factor-α were measured during the ischemia-reperfusion process.Results In comparison to adult lungs,the infant lungs had more increased neutrophil infiltration,edema,swelled alveolar epithelial and endothelial cells,and severer mitochondrial impairment reflected by damage of the inner membrane as well as decrease in the membrane potential after ischemia-reperfusion.The lungs in infant animals subjected to sham operation displayed higher levels of ROS-HR and MDA and lower levels of SOD and GSH-PX than those in adult controls.The lungs in infants with ischemia-reperfusion were found to further produce more ROS-HR,and MDA,and less SOD and GSH-PX than the ischemia-reperfused adult lungs.Moreover,the circulating levels of interleukin-1β and tumor necrosis factor-α were elevated during the period of ischemia-reperfusion,particularly in the infant animals,which appeared to be associated with the expression of MyD-88 and NF-κB in the lungs.Conclusion Lung ischemia-reperfusion causes more severe lung damage in infants than in adults,probably due to combination of low antioxidant capacity and overproduction of ROS in infants.
6.Effects of Ulinastatin and 1, 6 fructose diphosphate on GSH-Px and MDA in cerebral ischemia-reperfusion injury
Jian ZHANG ; Sheng-Tao YAN ; Shi-Dong GUO ; Li-Da ZHI ;
Chinese Journal of Emergency Medicine 2006;0(06):-
Objective To investigate the protective effects of Ulinastatin (UTI) and 1, 6 fructose diphosphate (FDP) on cerebral ischemia-reperfusion injury, and to discuss the protective mechanisms of UT1 and FDP on cerebral ischemia-repeffusion injury. Method Rabbit ischemia-repeffusion injury models were prepared by"four arteries occlusion". All rabbits were divided into four groups (n=6): control group, UTI (10 U/kg) group, FDP (200 mg/kg) group, UTI+ FDP group. Salt water, UTI, FDP and UTI + FDP were respectively used immediately after ischemia-reperfusion. Plasmic MDA and GSH-Px were measured at 30 minutes, 1 hour, 3 hours and 6 hours after reperfusion. Results The concentrations of plasmic MDA in every group were significantly improved compared with those in control group (P0.05 ), but significant difference could be found after 3 hours (P
7.Soluble Expression and Purification of Snake Venoms Fihrino(geno)lytic Emzyme Alfimeprase in E.coli
Shou-Tao ZHANG ; Yan-Sheng ZHOU ; Xue-Hua LAI ; Xing-Feng BAO ; Ai-Guang GUO ;
China Biotechnology 2006;0(03):-
Fibrolase is a non-hemorrhagic zinc metalloproteinase isolated from southern copperhead snake venom (Agkistrodon contortrix contortrix) and is capable of degrading fibrin clots resulting from purified fibrinogen or from blood plasma. Alfimeprase, a truncated form of fibrolase, as a clinical agent was successfully completed PhaseII clinical trials.The cDNA of alfimeprase was amplified by recursive PCR, digested with BamHI and HindII, and cloned into pET43.1a, pMALp2X and pMALc2X vectors to generate fusions with NusA, MBP and sMBP(with signal peptide), respectively. Nus/alfimeprase was expressed in soluble form by co-expressing with chaperone FkpA and inducing with1mmol/L IPTG. The fusion protein accounted for about 25 % of total protein following cell lysis. Alfimeprase was successfully purifiesd by Ni-NTA affinity chromatography and cleaved by enterokinase. The results demonstrate the fibrinolytic activity of recombinant alfimeprase using fibrin plate assays and fibrinogen hydrolysis.
8.Effect of carbonated hydroxyapatite cement for filling vertebral body on the vertebral heights and pain in patients with osteoporotic vertebral compression fractures
Keya MAO ; Baowei LIU ; Yan WANG ; Sheng TAO ; Jifang WANG ; Zhengsheng LIU ; Songhua XIAO ; Yonggang ZHANG
Chinese Journal of Tissue Engineering Research 2007;11(1):188-190,封3
BACKGROUND: Carbonated hydroxyapatite cement (CHC) s a new kind of biomaterial for bone defect, which is made of powder and fluid, and can be mixed to be pasty to repair various bone defects.OBJECTIVE: To observe the improvement of vertebrae height and pain in patients with osteoporosis vertebral compression fracture (VCF) after vertebroplasty by using a new kind of bone graft biomaterial, taking CHC as the filling material to reinforce the vertebral body.DESIGN: A contrast observation trial taking patients as subjects.SETTING: Department of Orthopaedics, General Hospital of Chinese PLA.PARTICIPANTS: Totally 34 patients with thoracic or lumbar osteoporosis VCF who received the treatment in the Department of Orthopaedics, General Hospital of Chinese PLA between October 2000 and August 2003. Inclusive criteria: ①Definite diagnosis by CT; ② Informed consents were obtained from the patients. Exclusive criteria: The patients with osteoporosis vertebral compression fractures who suffered vertebral posterior wall fracture. There were 6 males and 28 females, and they were aged (72±13)years; Among the patients, 27 were diagnosed as postmenopausal osteoporosis, 1 as cortical hormone-induced osteoporosis and 6 male patients weresenile osteoporosis.METHODS: ①All the patients were randomly divided into two groups: Experimental group (n =23) and control group (n=11). All the patients were performed percutaneous operation with local anesthenia. All cases were performed percutaneous operation under local anesthesia. Under the C-arm monitored, one side pedicle puncture was performed to enter the anterior column of the involved VCF. Patients of the experimental group were filled with CHC. Patients of control group were filled with polymethyl Methacrylate (PMMA) with the same way. ② Referred to McGill-Melzack scoring. Among the scale 0-100 mm (0 was no pain, 100 was acute pain), the value indicated the painful intensity and mental assault degree. < 30 scores indicated good, 30-40 basically satisfied and ≥ 50 poor .③ Referred to the method from Lee et al, the preoperative height (A1) and postoperative height (A2) of compression fracture position of VCF were measured according to the lateral X-ray film. At the same time, the upper vertebral height (A3) and the inferior vertebral height (A4) were measured at the same position. The original height (A) of the involved vertebra was calculated as (A)= (A3+A4)/2,and the preoperative vertebral compression rate =(A-A1 )/A, the postoperative vertebral compression rate =(A-A2)/A, the restoring rate = (the preoperative vertebral compression rate-the postoperative vertebral compression rate)/the preoperative vertebral compression rate. ④ The wounds of the patients were observed after operation. The levels of blood routine, serum calcium and serum phosphorus were detected before, one day and one week after operation. MAIN OUTCOME MEASURES: ① Preoperative and postoperative VAS scoring. ② The vertebral compression rate and restoring rate. ③ Wounds were observed after operation. The blood routine, the serum calcium and serum phosphorus were detected before, one day and one week after operation.RESULTS: Totally 34 patients were involved in the result analysis. ①The preoperative visual analogue scale (VAS) score of experimental group were (91.5±21.7) points, and the postoperative ones were (44.5±27.2) points. The difference of VAS score reduced gradually along with the postoperative time. There was no difference of VAS score between experimental group and the control group 4 weeks after operation. ② The biocompatibility of CHC in the vertebral body was fine. The vertebral compression rate of experimental group was recovered from (43.1±21.4)% preoperatively to (27.3± 18.5)% postoperatively. The rate of restored heights was (27.3±18.5)%. ③ All patients obtained Ⅰ stage wound healing, and none of them had infection, inflammatory secretion and nervous symptom. There were no differences in blood routine test, serum calcium, serum phosphorus between patients in two groups. One case filled by PMMA and two cases filled by CHC presented leakage, and none had nervous symptom.CONCLUSION: As the filling materials for vertebropalsty, CHC can restore the vertebral heights and relieve pain safely and effectively, however, its efficacy to relieve pain is not significant as PMMA in the short term.
9.Clinical anatomy of three major salivary glands to treat xerophthalmia
Xuan, XIAO ; An-huai, YANG ; Yan-ning, YANG ; Li, YU ; Sheng-xiang, TAO
Chinese Journal of Experimental Ophthalmology 2012;(10):889-892
Background Salivary transplantation or duct transposition can provide continuous physiological secretion of tear substitutes.This may be an ideal method in treatment of dry eye.But the relative anatomical literatures is few,and some of the conclusions in the literatures are still controversial,which limit its clinical application.Objective This study was to discuss the possibility and the advantage and disadvantage of applying three major salivary glands to treat xerophthalmia.Methods The relationship between the branches of the facial nerve out of the parotid gland and the salivary glands,the salivary glands size,origin of blood supply,out diameter of vessels and adjacent relation were observed in 34 sides pate specimens perfused with red latex under the operating microscope.To find the vessels in recipient site to anastomose,the vessels around fossa orbitalis and forehead were anatomized and observed.The parotid gland duct transfer operation,the submandibular gland free transplantation surgery and sublingual gland free transplantation surgery in the human anatomy specimens were simulated.Results The position of parotid duct was constant.The duct length was(4.20± 1.10) cm,duct diameter was (O.60±0.30) cm.The stensen's duct was likely to be prolonged by the cheek mucous membrane or venous andthe damage of buccal branch,zygomatic branch and temporal branches of facial nerve should be avoided during the operation of transplanting stensen' s duct.When submandibular gland was transplanted,facial vessel was taken as its pedicle,whose outside diameter was (2.70 ± 0.28) mm,and the length of the transplant vascular pedicle was (1.90 ± O.30) cm.Thc anastomosed vessel was superficial temporal vessel in recipient site.When sublingual gland was transplanted,sublingual(88.2%,30 sides) or submental vessel(11.8%,4 sides) was taken as its pedicle,whose outside diameter was(1.92±0.36) mm and (1.96±0.54) mm,and the length of the transplant vascular pedicle was(2.60± 1.10) cm and(3.50±0.40) cm,and the anastomosed vessel was the frontal branch of superficial temporal vessel in recipient site.Three sides of specimens lacked sublingual glands.Conclusions It is feasible that treating severe xerophthalmia by the operation of grafting the major salivary glands or transplanting stensen' s duct on the point of anatomical view.Parotid duct inversion and the submandibular gland transplantation have been applied to clinic.However,sublingual transplantation remains to be further confirmed by the animal experiments.
10.Puerarin effects on the mRNA expression of osteoblast differentiation-related proteins
Siyuan YUAN ; Beibei KONG ; Tong SHENG ; Xinxiang WANG ; Yunling ZHANG ; Xuemei LIU ; Tao MA ; Hong ZHENG ; Yan YAN ; Lianqi LIU
Chinese Journal of Tissue Engineering Research 2014;(42):6732-6736
BACKGROUND:Experimental studies have showed that puerarin has an obvious protective effect on osteoporosis in ovariectomized and orchiectomized mice. But the influence of puerarin in the molecular level in the process of osteoblast differentiation is seldom reported.
OBJECTIVE:To observe the effect of puerarin on the mRNA expression of alkaline phosphatase, bone sialoprotein, osteopontin and osteocalcin in osteoblasts.
METHODS:The MC3T3-E1 cells from mice cultured in vitro were randomly divided into control group, puerarin group (10-6 mol/L puerarin) and estradiol group (10-7 mol/L estradiol) to observe the effects of puerarin on the differentiation of osteoblasts. mRNA expression of alkaline phosphatase, bone sialoprotein, osteopontin and osteocalcin in MC3T3-E1 cells was determined using RT-PCR method.
RESULTS AND CONCLUSION:Puerarin and estradiol both could prolong the expression of alkaline phosphatase that reached the peak at 12 days. Puerarin and estradiol strengthened the mRNA expression of bone sialoprotein at 10 and 12 days, reduced expression of osteopontin at 5 and 12 days, and increased expression of osteocalcin at 10 and 12 days. These results reveal that puerarin can induce the differentiation of cultured osteoblasts by influencing osteoblast differentiation-related protein mRNA expressions, which may be one of the important molecular mechanisms of puerarin for prevention of osteoporosis.