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Objective:Rosin alcohol two glycosidase (pinores inoldiglucoside, PDG) effect on the improvement of the osteoporosis in mice and the possible mechanism of action.Methods:A mouse model of osteoporosis was established by removing bilateral ovaries. According to the random number table method, 50 mice were divided into model (MOD) group (equal volume of normal saline), positive control (PC) group (0.2 mg/kg estradiol), PDG-low dose (L) group (5 mg/kg) PDG), PDG-high dose (H) group (10 mg/kg PDG by gavage) and PDG-H+ML385 group (10 mg/kg PDG by gavage after intramuscular injection of 30 mg·kg-1 ML385), 10 mice in each group; Another 10 mice without any ovarian treatment were selected as sham group (intragastric administration of the same amount of normal saline). After 3 weeks of drug intervention, the serum alkaline phosphatase (ALP), tartrate resistant acid phosphatase (TRAP), interleukin-6 (IL-6) and tumor necrosis factor were detected by enzyme-linked immunosorbent assay (ELISA) -α (TNF-α). The levels of superoxide dismutase (SOD) and malondialdehyde (MDA) were measured by dual energy X-ray. The mRNA and protein expression levels of nuclear factor related factor 2 (Nrf2), oxidase heme oxygenase 1 (HO-1), quinone oxidoreductase 1 (NQO1) were detected by real-time fluorescence quantitative PCR (qRT-PCR) and Western blot, respectively.Results:The serum ALP levels in Sham, MOD, PC, PDG-L, PDG-H and PDG-H + ML385 groups were (25.88±3.42), (47.42±5.32), (36.20±3.37), (38.95±3.24), (29.66±2.64), (39.57±2.06) U/dL, respectively; The serum trap levels were (2.18±0.40), (4.69±0.83), (3.58±0.38), (3.67± 0.48), (2.93±0.26), (3.81±0.49) U/L, respectively; BMD (153.04±12.96), (86.25±6.71), (126.53±8.99), (119.77±8.84), (139.18±15.94), (92.09±4.17) mg/cm 2; The expression levels of Nrf2 mRNA were 1.00±0.00, 0.35±0.04, 0.67±0.05, 0.54±0.03, 0.82±0.08 and 0.48±0.04, respectively; The expression levels of HO-1 mRNA were 1.00±0.00, 0.25±0.03, 0.56±0.03, 0.47± 0.03, 0.71±0.04 and 0.37±0.04, respectively; The expression levels of nqo1mrna were 1.00±0.00, 0.38± 0.02, 0.63±0.03, 0.58±0.04, 0.79±0.05 and 0.44±0.03, respectively; Nrf2 protein expression levels were 0.98±0.08, 0.26±0.04, 0.52 ±0.06, 0.46±0.03, 0.86±0.07, 0.45±0.05. HO-1 protein expression levels were 0.39±0.02, 0.09±0.01, 0.15 ±0.03, 0.17±0.03, 0.26±0.03, 0.12±0.02. NQO1 protein expression levels were 0.53±0.03, 0.21±0.02, 0.38±0.04, 0.29±0.02, 0.55±0.03, 0.24±0.04, respectively; The levels of serum IL-6 were (104.25±11.35), (515.38±74.48), (351.78± 65.12), (364.73±36.64), (246.18±17.52), (408.93±32.56) pg/ml, respectively; The levels of serum TNF-α were (33.79±3.55), (170.11±19.24), (76.09±8.99), (84.95±6.12), (66.98±3.73), (119.04±9.75) pg/mL, respectively; The serum SOD levels were (258.47±19.25), (72.15±8.12), (187.60±14.63), (152.61±12.36), (204.22±19.65), (138.01±13.62) U/mL, respectively; The serum MDA levels were (2.02±0.27), (4.75±0.73), (3.19±0.46), (3.7±0.49), (2.91±0.42), (4.10 ±0.25) μmol/L respectively. There were significant differences between MOD group and Sham group, or PC, PDG-L, PDG-H, PDG-H+ML385 group and MOD group,or PDG-H+ML385 group and PDG-H ( P<0.05) . Conclusion:PDG can reduce bone inflammation and oxidative stress by activating Nrf2 pathway and improve the state of osteoporosis.
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ObjectiveTo explore the neuroprotective mechanism of Buyang Huanwutang （BYHW） on diabetic peripheral neuropathy （DPN） rats based on oxidative stress and investigate the dosage of Astragali Radix （AR）. MethodNinety SD rats were randomly divided into a normal group， a model group， an α-lipoic acid group （60 mg·kg-1·d-1）， and BYHW groups with high- （15 g·kg-1·d-1）， medium- （8.75 g·kg-1·d-1）， and low-dose （5.625 g·kg-1·d-1） AR groups. The diabetes model was induced in rats except for those in the normal group by the high-sugar/high-fat diet and intraperitoneal injection of streptozotocin （STZ）. Drug intervention lasted for 12 weeks. The paw withdrawal threshold （PWT） and sensory nerve conduction velocity （SNCV） were detected after drug intervention. Gonad-stimulating hormone （GSH） and malondialdehyde （MDA） were determined. The mitochondrial morphology and structure in sensory neurons of L4-5 dorsal root ganglion （DRG） of rats were observed by electron microscopy. Respiratory chain complex Ⅰ， Ⅱ， Ⅲ， and Ⅳ activities and the mitochondrial membrane potential were detected. The main proteins in the adenosine monophosphate-activated protein kinase （AMPK）/nuclear factor-related factor-2 （Nrf2） pathway， such as phosphorylated AMPK （p-AMPK）， phosphorylated Nrf2（p-Nrf2）， heme oxygenase-1 （HO-1）， and quinone NADH dehydrogenase 1 （NQO1）， were detected by immunohistochemistry and Western blot. ResultCompared with the normal group， the model group showed increased fasting blood glucose （P<0.01）， decreased content of SNCV， PWT， and GSH （P<0.01）， elevated MDA content （P<0.01）， obvious mitochondrial damage with vacuolations， reduced activities of respiratory chain complex Ⅰ， Ⅱ， Ⅲ， and Ⅳ and mitochondrial membrane potential （P<0.01）， and declining p-AMPK， p-Nrf2， HO-1， and NQO1 （P<0.01）. Compared with the model group， the α-lipoic acid group and BYHW high-dose group showed increased SNCV， PWT， and GSH， decreased MDA （P<0.05， P<0.01）， alleviated mitochondrial structural damage， increased respiratory chain complex Ⅰ， Ⅱ， Ⅲ， and Ⅳ activities and mitochondrial membrane potential （P<0.01）， and elevated p-AMPK， p-Nrf2， HO-1， and NQO1 （P<0.05， P<0.01）. ConclusionBYHW regulates oxidative stress through the AMPK/Nrf2 pathway to treat DPN. The therapeutic effect of BYHW is related to the dosage of AR. The BYHW group with high-dose AR is superior to the BYHW groups with medium- and low-dose AR groups in inhibiting oxidative stress.
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Objective To investigate the correlation between nuclear factor-associated factor 2 gene polymorphism and COPD and the effect of tiotropium bromide combined with budesonide/formoterol inhalation. Methods Seventy-nine patients with chronic obstructive pulmonary disease and 54 healthy subjects were enrolled to determine the NRF2-617C/A(rs6721961)polymorphism by PCR-RFLP and to analyze the relationship between different genotypes and COPD ,and then to observe the therapeutic effect of different genotypes. Results The frequency of A allele in NRF2-617C/A(rs6721961)was a risk factor for COPD(OR=2.312,95%CI:1.517 to 6.824,P<0.05). The total effective rate of AA was lower than that of CC and CA(χ2=8.65,7.24,P<0.05). After treatment,FEV1% pred,FEV1/FVC in the AA type were significantly lower in the CC type(t=6.48,5.62, P<0.05),while IL-8,TNF-αand hs-CRP were significantly higher than those in the CC type(t=6.07,9.21, 4.79,P<0.05). Conclusion NRF2-617C/A(rs6721961)gene polymorphism has a certain correlation with COPD,on which it has a significant effect to tiotropium combined with budesonide treatment and has a moderating effect on inflammatory response and pulmonary function in COPD patients.