1.Analysis on Volatile Components in Cuscuta chinensis from Different Habitats by HS-SPME-GC-MS
Xuejun PEI ; Jinqing LU ; Qiang LI ; Shengnan GUO
China Pharmacy 2016;27(21):3006-3008,3009
OBJECTIVE:To establish a method for analyzing the volatile components in Cuscuta chinensis,and compare the difference of the volatile components in C. chinensis. METHODS:HS-SPME-GC-MS was adopted:sampling amount was 1.0 g, extracting fibers was 65 μm PDMS/DVB,equilibrium temperature was 120 ℃,equilibrium time was 15 min,extraction time was 30 min,resolution time was 3 min;GC conditions:the column was HP-5MS quartz capillary column,programmed temperature, inlet temperature was 230 ℃,carrier gas was high purity helium,the flow rate was 1.0 ml/min,splitless injection;MS condi-tions:ion source was electron ionization,temperature was 230 ℃,quadrupole temperature was 150 ℃,electron energy was 70 eV,photomultiplier tube voltage was 1.2 kV,the interface temperature was 280 ℃,and scanning range was m/z 35-550. Com-bined with the qualitative analysis for volatile components of C. chinensis from different habitats by HP ChemStation,the relative content was calculated by peak area normalization,and the data was analyzed by principal component analysis and cluster analysis. RESULTS:Totally 52 components were identified,9 of which were the common components in C. chinensis,namely leaf alcohol, 1-octene-3-ol,3-octanol,malt alcohol,diethyl phthalate,caryophyllene,nonaldehyde,octanol and palmitic acid. sample 1,2,3 were clustered into a group,then clustered with 4,5,6 into a group,sample 7,8,9 was clustered into a group,then clustered with 10,11,12 into a group,and sample 13,14,15 clustered into a group individually. CONCLUSIONS:The method is stable and reliable,and suitable for the rapid analysis of volatile components in C. chinensis;and differences of volatile components in C. chinensis from diflerent habitats are discernible.
2.GC-MS Comparative Analysis of Volatile Compounds Extracted from Rhodiola Crenulata Respectively by HS-SPME and SD
Shengnan GUO ; Jinqing LU ; Junlong CAI ; Qiang LI ; Huan LIANG
China Pharmacist 2014;(11):1885-1888
Objective: To compare the volatile compounds extracted from Rhodiola renulata respectively by HS-SPME and SD. Methods:The volatile constituents from Rhodiola crenulata were extracted respectively by HS-SPME and SD, and then the contents and the names were confirmed by GC-MS. Results:Totally 39 compounds were identified from Rhodiola crenulata by HS-SPME while 16 ones were identified by SD. Among them, 4 common compounds were detected. Conclusion: There are some differences between the two methods. Compared with SD, HS-SPME is obviously better because more volatile constituents can be extracted from the herb, furthermore, HS-SPME has notable advantages of higher retrieval matching and sensitivity.
3.Antioxidant Activity of Total Flavonoids of Cinnamomum Septentrionale Hand.-Mzt
Hua LU ; Yisheng ZHANG ; Shengnan GUO ; Qiang LI ; Yuling LI ; Jinqing LU
China Pharmacist 2015;(4):555-557
Objective:To investigate the antioxidant activity of the total flavonoids of Cinnamomum septentrionale Hand.-Mzt. Methods:A ferricyanide reduction method was used to study the reduction ability. The scavenging activity of the total flavonoids on DPPH· free radical( DPPH·),hydroxyl radicals( ·OH)and superoxide anion free radical( O2-·)was investigated as well. Re-sults:Cinnamomum septentrionale Hand.-Mzt. had good reduction and scavenging activity on the free radicals in a concentration-de-pendant manner. When the concentration of total flavonoids of Cinnamomum septentrionale Hand.-Mzt reached 0. 15 mg·ml-1 ,86%of DPPH free radical( DPPH·),68% of hydroxyl radicals( ·OH)and 59% of ultra oxygen anion free radical( O2-·)were scav-enged. Conclusion:Cinnamomum septentrionale Hand.-Mzt. has good antioxidant activity,and as a natural antioxidant,it is worthy of further research and development.
4.Effect of astragaloside IV on expression of PGC-1αand NRF-1 in myocardial cells of typeⅠdiabetic rat
Qiongdan CAO ; Yuhong YANG ; Shengnan YU ; Meili LU ; Suping ZHANG ; Ronghui HAN ; Jin HU
Chinese Pharmacological Bulletin 2015;(8):1096-1100,1101
Aim To investigate the effect of astragalo-side IV ( ASIV) on myocardial energy metabolism and mitochondrial biosynthesis in myocardial cells of dia-betic rats induced by streptozotocin ( STZ ) . Methods
50 SD rats at 6 weeks of age were assigned to 5 groups,10 for each group:control group, model group, ASIV 10 mg·kg-1 ·d-1 group, ASIV 20 mg·kg-1 ·d-1 group, ASIV 40 mg·kg-1 ·d-1 group. Except the control group,the remaining 40 were used to estab-lish type 1 diabetes model by the tail vein injection of STZ (35 mg·kg-1 ) . At the end of 16 weeks of treat-ment, left ventricular systolic pressure ( LVSP ) , left ventricular diastolic final pressure ( LVEDP ) and left ventricular maximum rising/falling rate ( ± dp/dtmax ) were tested. Pathological section was observed by HE staining. ATP, ADP, AMP levels were detected by ELISA. The expressions of PGC-1α and NRF-1 protein were assessed by Western blot. The expressions of PGC-1α and NRF-1 mRNA were determined by RT-PCR. Results Compared with control group, model group markedly elevated LVEDP and decreased LVSP, ± dp/dtmax , ATP/AMP and ATP/ADP ratio. Com-pared with model group, low-dose ASIV group did not change significantly,middle-dose ASIV group and high-dose ASIV group obviously decreased LVEDP, and im-proved LVSP, ± dp/dtmax , ATP/ADP and ATP/AMP ratio. Meanwhile, the expressions of PGC-1α and NRF-1 protein and mRNA were increased in a dose-de-pendent manner. Conclusion ASIV could promote mitochondrial biosynthesis, improve energy metabolism in myocardial cells of type 1 diabetic rats by PGC-1αand NRF-1 .
5.Effect of meloxicam on CUMS-induced depressive-like behavior in rats and its preliminary mechanism
Shengnan KUANG ; Ying LUO ; Xiaoyan TIAN ; Lu ZHANG ; Yang YANG ; Junqing YANG
Chinese Pharmacological Bulletin 2016;(2):263-267,268
Aim To explore the effect of meloxicam on the CUMS-induced depressive-like behaviors in rats and its preliminary mechanism. Methods The rats were exposed to CUMS procedure for 6 weeks to estab-lish the model of depression. Meloxicam(1,3 mg· kg-1 ) and sertraline(5 mg·kg-1 ) were administered to rats from 22d of the stress procedure(once a day,for 21 days,p. o. ) . Depressive-like behaviors were evalu-ated by the open-field test and force swimming test. The levels of PGE2 and TNF-αin cortex were measured by ELISA. Moreover, the concentrations of NE, DA, DOPAC and 5-HIAA were also measured by HPLC, and the protein expression of 5-HT1 AR in cortex was analyzed by the immunohistochemistry. Results Com-pared with the rats of normal control group,the vertical and horizontal movement scores of rats in the open-field test were decreased and the immobility time in the forced swimming test was increased in model group. The levels of PGE2 and TNF-α were both increased signifi-cantly,whereas the concentrations of NE, DA, DOPAC and 5-HIAA were decreased and the expression of 5-HT1AR was reduced in cortex. Compared with the rats of model group, meloxicam significantly improved the depressive behaviors of rats in experimental groups and reversed the content of PGE2 ,TNF-α,NE,DA,DOPAC and 5-HIAA, as well as the expression of 5-HT1AR. Conclusion Meloxicam has a significant protective effect on CUMS-induced depressive-like behaviors, and the protective mechanism might be related to atten-uating inflammation response and reconstructing the balance of the monoamine neurotransmitter system in rat cortex.
6.Recombination of RegIII-proinsulin-pBudCE4.1 plasmid and its therapeutic effect on STZ-induced type 1 diabetes mellitus.
Wenrui HOU ; Shengnan XIE ; Jingli LU ; Wei XI ; Xiang LUO ; Ming XIANG
Acta Pharmaceutica Sinica 2010;45(8):987-94
The aim of this study is to investigate the therapeutic effect of RegIII-proinsulin-pBudCE4.1 plasmid on streptozotocin (STZ)-induced type 1 diabetes mellitus and its underlying mechanisms. The model of type 1 diabetes mellitus was established by intraperitoneal injections of STZ (40 mg kg(-1)) to Balb/c mice for five consecutive days. Then, ten type 1 diabetic mice were intramuscularly injected with 100 microg RegIII-proinsulin-pBudCE4.1 plasmid for 4 weeks (one time/week) and the blood glucose levels were monitored every week; whereas another ten diabetic mice served as negative control group were injected with pBudCE4.1 vector at the same dose. Normal control and model control mice were treated with normal saline at identical volume under the same way. Western blotting, MTT assay, ELISA, HE staining and Tunel assay were applied to explore the underlying mechanisms. Results showed that RegIII-proinsulin-pBudCE4.1 plasmid ameliorated the hyperglycemia symptoms in diabetic mouse remarkably. It induced an immunological tolerance state in type 1 diabetic mice by inhibiting the proliferation of splenic lymphocytes and recovering Th1/Th2 balance evidenced by MTT and ELISA analysis. Furthermore, it elevated insulin concentration in the serum of type 1 diabetic mice and promoted the regeneration of beta cells supported by the results of HE staining and Tunel assay. In conclusion, RegIII-proinsulin-pBudCE4.1 plasmid possesses powerful anti-diabetic ability, which may be involved in the inducing of immunological tolerance and enhancing beta cells recovery.
7.Effect of Tripterygium wilfordii combined with methotrexate on short-term effect of rheumatoid arthritis and the levels of IL-6, IL-10 and TNF- in peripheral blood
Shengnan WANG ; Ping CHEN ; Suxian LIN ; Jianchang JIANG ; Yang LU ; Zhiyong ZHANG ; Xiaochun ZHU
Chinese Journal of Biochemical Pharmaceutics 2017;37(9):202-204
Objective To investigate the effects of tripterygium glycosides combined with methotrexate in treatment of rheumatoid arthritis (RA) curative effect and peripheral blood of patients with IL-6, effects of IL-10 and TNF- alpha. Methods 80 patients with RA from February 2014 to August 2016 in our hospital were retrospectively analyzed. According to the different treatment methods by for the observation group, the control group (40 cases). To compare and analyze the curative effect before and after short-term treatment and treatment of patients in the 2 groups were the main clinical symptoms and signs of change and ESR, RF, CRP, IL-6, TNF- alpha, IL-10 index. Results The total effective rate of observation group (92.50%) was significantly higher than the control group (77.50%), the difference was statistically significant (P<0.05), 2 groups of patients before treatment of main clinical symptoms and signs, there was no significant difference, after treatment, clinical symptoms and signs of 2 groups of patients were compared with those before treatment was statistically significant to alleviate the differences (P<0.05), and the observation group of main clinical symptoms and signs were lower than the control group and the remission (down), the difference was statistically significant (P<0.05).Before ESR, the treatment of 2 patients with RF, CRP, TNF- IL-6, alpha, IL-10 index level, no statistical significance, the 2 groups after treatment in patients with ESR, RF, CRP, IL-6, TNF- alpha, IL-10 index level than before treatment with significant difference (P<0.05), and the observation group ESR, RF, CRP, TNF- alpha, IL-6 average index of water compared with the control group decreased significantly, IL-10 index increased significantly than the control group. The difference was statistically significant (P<0.05). Conclusion Tripterygium glycosides combined with methotrexate in treatment of rheumatoid arthritis And it can effectively inhibit the expression of peripheral serum L-6 and TNF- alpha, promote the expression of IL-10 and alleviate the inflammatory reaction
8.Effects of eplerenone on expression and activity of aortic endothelial nitric oxide synthase in hypertensive rats induced by high-salt intake
Qian ZHANG ; Jing DING ; Fei YANG ; Shengnan WANG ; Feng SU ; Deqin LU
Chinese Journal of Pathophysiology 2015;(9):1606-1610
[ ABSTRACT] AIM:To explore the effects of eplerenone on the expression and activity of aortic endothelial nitric oxide synthase ( eNOS) in high salt-induced hypertensive rats .METHODS: Male Wistar rats (4 week old, weighting 50~60 g) were randomly divided into control group , high-salt diet group and eplerenone group .The rats in control group were fed with ordinary rodent animal diet , the rats in high-salt group and eplerenone group were exposed to 5%salt diet for 16 weeks and administrated with the same dosage of saline or eplerenone (40 mg? kg-1? d-1 ) by gavage for 4 weeks, re-spectively.Systolic blood pressure (SBP) was measured by tail-cuff every 2 weeks.The rats were sacrificed after 16 weeks and the thoracic aorta was collected .The aldosterone content in the aorta was measured by ELISA .The protein levels of mineralocorticoid receptor (MR) and eNOS were determined by Western blot.The activitie of constitutive NOS (cNOS) was measured by chemocolorimetry .The protein localization of eNOS , neuronal nitric oxide synthase ( nNOS) and MR was observed by immunohistochemistry .RESULTS: A process of 8-week high-salt diet increased SBP gradually .SBP in the rats exposure to high salt for 16 weeks was significantly higher than that in control group ( P<0.05 ) .After 4 weeks of eplerenone treatment, SBP in the rats was significantly lower than that before treatment (P<0.05).Compared with control group, the aldosterone content in the aorta were significantly increased in high-salt diet group and eplerenone group ( P<0.05), the expression level of MR also increased significantly (P<0.05).Compared with control group, both eNOS pro-tein expression (P<0.05) and cNOS activity in high-salt diet group were significantly decreased (P<0.05).The protein expression of eNOS as well as cNOS activity in aorta increased significantly in eplerenone group compared with high -salt diet group (P<0.05).CONCLUSION:Aldosterone content in aorta of high-salt-induced hypertensive rats increases signifi-cantly .Aldosterone attenuates the protein expression of eNOS and reduces the enzyme activity through the activation of min -eralocorticoid receptor .The selective mineralocorticoid receptor antagonist eplerenone enhances the protein expression of eNOS and its activity , thereby improves eNOS function .
9.Ablation of adenosine monophosphate-activated protein kinaseα1 in vascular smooth muscle cells promotes diet-induced atherosclerotic calcification in vivo
Zhejun CAI ; Ye DING ; Miao ZHANG ; Qiulun LU ; Shengnan WU ; Huaiping ZHU ; Ping SONG ; Minghui ZOU
Chinese Journal of Pathophysiology 2016;32(8):1493-1493,1494
AIM:Atherosclerotic calcification is highly linked with plaque instability and cardiovascular events .Adenosine monophosphate-activated protein kinase ( AMPK) has been involved in the pathogenesis of various cardiovascular disease .The contributions of AMPKαsubunits to the development of atherosclerotic calcification in vivo remained unknown .We hypothesized that AMPKαsubunits may play a role in the development of atherosclerotic calcification .METHODS: Atherosclerotic calcification was generated by 24-week fed of western diet in ApoE-/-background mice .Calcification was evaluated in aortic roots and innominate arteries of ApoE-/-mice or in mice with dual deficiencies of ApoE and AMPKαsubunits globally ( AMPKα1 and AMPKα2 ) , or vascular smooth muscle cell ( VSMC)-specific or macrophage-specific knockout of AMPKα1 with atherosclerotic calcification pone diet . The mechanism of AMPKα1 in regulating Runx2 was further explored in human aortic VSMC .RESULTS: Ablation of AMPKα1 but not AMPKα2 in ApoE-/-background promoted atherosclerotic calcification with increased Runt -related transcription factor ( Runx2 ) expression in VSMC compared with ApoE-/-mice.Conversely, chronic administration of metformin, which activated AMPK, markedly reduced ath-erosclerotic calcification and Runx2 expression in ApoE-/-mice but had less effects in ApoE-/-/AMPKα1 -/-mice.Furthermore, VSMC-but not macrophage-specific deficiency of AMPKα1 in ApoE-/-background promoted atherosclerotic calcification in vivo com-pared with the controls .AMPKα1 silencing in human aortic VSMC prevented Runx 2 from proteasome degradation to trigger osteoblastic differentiation of VSMC .Conversely , activation of AMPK led to Runx 2 instability by inducing its small ubiquitin-like modifier modifi-cation (SUMOylation).Protein inhibitor of activated STAT-1 (PIAS1), the SUMO E3-ligase of Runx2, was directly phosphorylated by
AMPKα1 at serine 510, to enhance its SUMO E3-ligase activity.Ablation of PIAS1 serine 510 phosphorylation inhibited metformin-in-duced Runx2 SUMOylation, and subsequently prevented the effect of metformin on reducing oxLDL-triggered Runx2 expression in hu-man aortic VSMC.CONCLUSION:Deficiency of AMPKα1 in VSMC increases Runx2 expression and promotes atherosclerotic calcifi-cation in vivo.AMPKα1 phosphorylates PIAS1 to enhance Runx2 SUMOyalation and subsequent degradation .
10.The study on the combined regional homogeneity and functional connectivity of resting-state magnetic resonance in patients with subjective cognitive decline
Tenglong WANG ; Xingfu ZHAO ; Yue WU ; Zaohuo CHENG ; Zhenhe ZHOU ; Dong WANG ; Liping LU ; Xuean WU ; Shengnan HAN
Chinese Journal of Geriatrics 2021;40(1):72-75
Objective:To investigate the mechanism of subjective cognitive decline(SCD)in resting-state by using regional homogeneity(ReHo)and functional connectivity(FC)in SCD patients.Methods:Resting-state functional magnetic resonance imaging(RS-fMRI)was performed in 25 SCD patients and 30 normal controls matched by sex, education and nationality.DPARSFA2.3 and SPM8 software were used to analyze and screen the brain areas with abnormal ReHo values in SCD group, with the posterior cingulated(PCC)/paruneus as seed points for whole-brain FC analysis.Results:Compared with the normal control group, the SCD group showed that ReHo values of right occipital gyrus and left precuneus were increased, and ReHo values of right inferior temporal gyrus, right orbital inferior frontal gyrus and bilateral thalamus were decreased(Voxel level, Alphasim correction, P<0.05). Using PCC/ precuneus as seed voxels, the whole brain functional connectivity analysis showed that the functional connectivity with cerebelum Crus 2 R was increased, and the functional connectivity with right orbital inferior frontal gyrus, left inferior temporal gyrus and temporal pole was reduced(Voxel level, Alphasim correction, all P<0.05). Conclusions:Default mode network may play an important role in the mechanism of SCD, and abnormalities in brain areas may first occur in PCC/precuneus.