1.CRMP2 alleviates neurological deficit by reducing neuron apoptosis in rats after cerebral ischemia/reperfusion injury
Xiangfeng XING ; Tianzhu WANG ; Xinyue QIN
Chinese Pharmacological Bulletin 2016;32(4):548-553
Aim To investigate the influence of the overexpression of CRMP2 on neural cell apoptosis after ischemia reperfusion injury in rats and its possible mechanism. Methods A total of 192 male adult SD rats were divided into four groups: sham group, cere-bral ischemia/reperfusion group( MCAO group) , cere-bral ischemia with blank plasmid control group( MCAO+GFP group ) , cerebral ischemia with CRMP2 eu-karyotic plasmid group ( MCAO + CRMP2/GFP group) . One day after injecting eukaryotic plasmid, the rats were operated for 120-min ischemia through MCA occlusion and reperfused. At 48 h and 1 wk, the expression of CRMP2 , p53 , Caspase-3 , Caspase-8 and BCL2 in brain tissue was tested by RT-PCR and West-ern blot. Apoptotic cells were observed by TUNEL test. TTC staining was use to detect cerebral infarction volume. The neural function of the rats were also eval-uated. Results Compared with the sham group, the expression levels of CRMP2 and BCL2 in MCAO group and MCAO +GFP group were significantly decreased ( P <0. 01 ) , while p53 , Caspase-3 , Caspase-8 and TUNEL positive cells were elevated(P<0. 01). Inter-vention of CRMP2 eukaryotic plasmid resulted in the increased expression of CRMP2 and BCL2 ( P<0. 01 ) and the decreased p53 , Caspase-3 and Caspase-8 ex-pression. In TUNEL test, overexpression of CRMP2 obviously decreased the number of TUNEL positive cells(P<0. 01). The expression of BDNF was upregu-lated after cerebral ischemic injury ( P<0. 01 ) , while overexpression of CRMP2 increased BDNF more signif-icantly ( P <0. 01 ) . TTC staining showed cerebral in-farction Volume of MCAO + CRMP2/GFP group was obviously decreased ( P <0. 01 ) , and neurologic defi-cits were significantly improved ( P <0. 01 ) . Conclu-sion The overexpression of CRMP2 reduces nerve cell apoptosis possibly by regulating the mitochondrial ap-optosis pathway after cerebral ischemia/reperfusion in-jury to protect nervous system.
2.Pioglitazone administration combined with bone marrow mesenchymal stem cells transplantation improved the heart function of rats with myocardial infarction
Quanhua WU ; Jingying HOU ; Tianzhu GUO ; Tingting ZHONG ; Huibao LONG ; Yue XING ; Changqing ZHOU ; Shaoxin ZHENG ; Tong WANG
Chinese Journal of Tissue Engineering Research 2015;(23):3698-3704
BACKGROUND:Our previous work has demonstrated that bone marrow mesenchymal stem cels (BMSCs) transplantation can improve the heart function of rats with myocardial infarction. However, the overal efficacy is not satisfactory. OBJECTIVE: To adopt pioglitazone as a peroxisome proliferator-activated receptor gamma (PPAR-γ) agonist combined with BMSCs transplantation therapy, thereby further improving cardiac function of rats with myocardial infarction as wel as investigating the relevant mechanisms. METHODS:Twenty Sprague-Dawley rats with myocardial infarction were induced by the left anterior descending coronary artery ligation. The animals were randomized into two groups: BMSCs and BMSCs+pioglitazone. Two weeks later, al the animals received the injection of BMSCs labeled with PKH26 in PBS into the local infarct zone, and then pioglitazone (3 mg/kg/d) was given by the oral gavage for 2 weeks in the BMSCs+pioglitazone group after the cel transplantation. After 2 weeks of cel transplantation, cardiac functions were evaluated by echocardiography. The expressions of PPAR-γ, Connexin 43 and molecules in TGF-β1/SMAD signaling pathway were examined in different areas of the left ventricle from each harvested heart using immunofluorescent staining, western blot assay and qRT-PCR. RESULTS AND CONCLUSION:There were no differences in the baseline parameters of cardiac function between the two groups. At 2 weeks after cel transplantation, the left ventricular internal diameter at end-diastole, left ventricular internal diameter at end-systole and left ventricular ejection fraction were significantly improved in the BMSCs+ pioglitazone group; the expressions of PPAR-γ and Connexin 43 were distinctly increased in different zones of the left ventricle; the levels of TGF-β1, SMAD2 and SMAD3 were obviously attenuated in the infarct zone and border zone. The above-mentioned findings suggest that pioglitazone, a PPAR-γ agonist, can enhance BMSCs potential in improvingthe heart function after myocardial infarction, and PPAR-γ may elevate the expression of Connexin 43via the blockade of the TGF-β1/SMAD signaling pathway in the procedure.
3.Study on Virtual Screening of Active Components from Yiqi Ziyin Decoction in the Treatment of Type 2 Diabetes Mellitus
Tianzhu XING ; Meng XU ; Zibo LIU ; Xiaojuan YU ; Shun XU
China Pharmacy 2019;30(7):910-916
OBJECTIVE: To conduct virtual screening for active component, target and pathway of TCM Yiqi ziyin decoction in the treatment of type 2 diabetes mellitus, and to elucidate the pharmacodynamic mechanism of this decoction in the treatment of type 2 diabetes mellitus at molecular level so as to provide theoretical basis for its popularization, application and rational organization. METHODS: Chemical compounds isolated from 10 ingredients of Yiqi ziyin decoction were retrieved from Handbook of Chemical Constituents of Original TCM Plants. Drug/drug-like molecules for the treatment of type 2 diabetes mellitus were retrieved from DrugBank database. Three-dimensional molecular structures of each component were downloaded from PubChem Compound database. The structure was optimized by using Cerius2 4.10 software, and the small molecular data set of Yiqi ziyin decoction was established. Using “type 2 diabetes mellitus” as keywords, target protein of type 2 diabetes mellitus were retrieved and screened from target protein databases, and their three-dimensional structures were downloaded from protein database. The selected compound molecules (ligands) were docked with target proteins (receptors), and small molecules with high activity (sorted according to the values of Degree and Betweenness) and target proteins with high correlation were found. The interaction mechanism between ligand and receptor was discussed by taking the fine docking of selected small molecules with target protein (N-trans-caffeoyl tyrosine molecule and glycogen synthase kinase-3β) as an example. RESULTS: There were 732 kinds of chemical components and 127 drug/drug-like molecules; 30 target proteins of type 2 diabetes mellitus were screened. According to the values of Degree and Betweenness, 8 molecules with high activity (such as N-trans-caffeoyl tyrosine, hyperin and so on) and 10 target proteins with high correlation (such as carbonic anhydrase1, glycogen synthetase kinase-3β and so on) were screened out. The interaction between N-trans-caffeoyl tyrosine and glycogen synthase kinase-3β could form hydrogen bonds and π alkylation, thus affecting the structural stability and activity of the latter and reducing blood glucose. CONCLUSIONS: In this study, it found there existed not only one molecule compound interacting with multiple targets, but also different molecule compound interacting with the same target on Yiqi Ziyin decoction for the treatment of type 2 diabetes mellitus by virtual screening, it revealed the characteristics of multi-component and multi-target synergism of TCM prescriptions at the molecular level, providing reference for screening lead compounds and structural modification of new drug for diabetes mellitus.