1.Effect of lactuside B on the expression of bcl-2 and bax mRNA and their protein in rats' cerebral cortex after cerebral ischemia-reperfusion injury.
Shengying LI ; Juan SUN ; Bingxuan NIU ; Fulin YAN ; Heqin ZHAN
Acta Pharmaceutica Sinica 2011;46(11):1314-20
This study is to investigate the effect of the major chemical composition in rhizome of Pterocypsela elata, lactuside B, on expression of bcl-2, bax mRNA and their protein in rats' cerebral cortex after cerebral ischemia-reperfusion injury. First, middle cerebral artery ischemia-reperfusion injury model was established, and each group was treated with the corresponding medicines. Animals were separately sacrificed at 24 h and 72 h. The brain infarct volumes were detected by TTC dye, bcl-2 and bax mRNA expression was checked by RT-PCR, and the proteins of bcl-2 and bax were explored by two-step immunohistochemistry in cerebral cortex of rats. Lactuside B can reduce brain infarct volume of cerebral cortex of rats, increase the expression of bcl-2 mRNA and decrease that of bax mRNA. Moreover, the ratio of bcl-2 to bax mRNA is higher in 12.5 and 25 mg kg(-1) dose group, respectively, which is significantly different from that of model group (P < 0.05 or P < 0.01). Generally, either 12.5 or 25 mg kg(-1) dose group is better than positive control medicine nimodipine (P < 0.05 or P < 0.01). In addition, the expression of bcl-2 and bax protein is consistent with their gene expression. Infarct volume and the ratio of bcl-2 to bax mRNA expression are significantly different (P < 0.05 or P < 0.01) between 72 h and 24 h group. The results demonstrated that lactuside B could play a good role in resisting cerebral ischemia by upregulating the expression of bcl-2 mRNA and protein and downregulating that of bax mRNA and protein.
2.Inmbition of nodosin on cell proliferation and expression of Bcl-2 and Bax in HepG2 cells
Guangfan HAI ; Bingxuan NIU ; Pinpin LI ; Lanqing GUO ; Taizhen CUI ; Shengying LI
Chinese Journal of Pathophysiology 2014;(10):1879-1882
AIM:To investigate the effects of nodosin extracted from Chinese traditional medicine on the pro-liferation of HepG2 cells cultured in vitro and to detect the protein expression of Bcl-2 and Bax in HepG2 cells.METH-ODS:HepG2 cells were treated with different concentrations (1.25, 2.5, 5, 10 and 20 μmol/L) of nodosin for 24 h. The morphological changes of HepG2 cells were observed under inverted microscope.The inhibitory rates of HepG2 cell growth were detected by MTT assay.The apoptotic rates and the protein expression of Bcl-2 and Bax were analyzed by flow cytometry.RESULTS:Shrunken and suspended HepG2 cells increased with the increases in the concentrations of nodo-sin.The apoptotic rates and the expression of Bax increased with the increases in the doses of nodosin, while the expression of Bcl-2 decreased.CONCLUSION:Nodosin inhibits the growth of HepG2 cells in a dose-dependent manner.The inhibi-tion of HepG2 cell growth is induced by decreasing Bcl-2 and increasing Bax, thus promoting cell apoptosis.
3.A study of Tongqiao Huoxue decoction combined with edaravone and hyperbaric oxygen for treatment of delayed encephalopathy after carbon monoxide poisoning
Xibo SUN ; Chuanlei CHEN ; Haiying PAN ; Chenghua ZHAO ; Zhen LI ; Mingde LIU ; Jianyi NIU ; Bingxuan LI
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2015;26(2):170-172
Objective To study the clinical curative effect of Tongqiao Huoxue decoction combined with edaravone and hyperbaric oxygen for treatment of delayed encephalopathy after carbon monoxide poisoning (DEACMP). Methods A prospective study was conducted. Forty-six patients with DEACMP admitted into Yidu Central Hospital of Weifang Medical College from January 2012 to January 2014 were randomly divided into observation group (23 cases) and control group (23 cases). The basic treatments of two groups were identical. Based on the basic treatments including hyperbaric oxygen and citicoline sodium injection etc, the observation group was treated with our-self made Tongqiao Huoxue decoction which could be modified in accord to the individual differentiation of syndromes in traditional Chinese medicine (the composition of decoction included Paeoniae Radix Rubra 15 g, Chuanxiong Rhizoma 15 g, Persicae Semen 15 g, Carthami Flos 15 g, Zingiberis Rhizoma Recens 3 pieces, Jujubae Fructus 2 pieces, Moschus 0.5 g, Allium Fistulosum 1 segment). The decoction was administered orally or by nasogastric gavage, one dosage everyday for 1 month, and in the mean time, edaravone intravenously drip 30 mg was given to the observation group twice a day for 14 days. The control group was given hyperbaric oxygen and other conventional treatment for 30 days. The clinical therapeutic effect and adverse reaction were observed after treatment for 30 days. The changes of intelligent level were detected by Hasegawa dementia scale (HDS), and the changes of latency of P300 were measured by electromyologram/evoked potential instrument in two groups before and after treatment. Results The total effective rate in observation group was significantly higher than that in control group [91.3% (21/23) vs. 65.2% (15/23), P < 0.01]. Elevation of creatinine occurred in 1 case, moderate increase in alanine aminotransferase (ALT) appeared in 1 case, and both of them were reduced to normal after treatment in observation group; no adverse reaction occurred in control group. The HDS scores were significantly higher 30 days after treatment than those before treatment in the two groups [control group:13.4±2.8 vs. 6.8±2.3, observation group:20.8±3.4 vs. 6.6±2.5, both P<0.05]. The latency of P300 after treatment was significantly lower in two groups than that before treatment [control group (ms): 355.7±25.7 vs. 385.5±27.8, observation group (ms): 337.3±24.6 vs. 386.8±25.4, both P < 0.05], the change in observation group being more significant [the HDS score: 20.8±3.4 vs. 13.4±2.8, the latency of P300 (ms): 337.3±24.6 vs. 355.7±25.7, both P<0.05]. Conclusion Tongqiao Huoxue decoction combined with edaravone and hyperbaric oxygen has favorable cognitive effect on patients with DEACMP, thus, it can be used extensively in clinic.
4.Rhein attenuates bleomycin-induced rats pulmonary fibrosis through TGF-β1/Smad pathway by inhibiting miR-21 expression
Yan QU ; Chong ZHANG ; Yanlong JIA ; Yu SONG ; Bingxuan NIU ; Heqin ZHAN
Chinese Journal of Pathophysiology 2017;33(1):149-153
AIM:To investigate the effect of rhein on bleomycin-induced pulmonary fibrosis and the expression of microRNA-21 (miR-21) and transforming growth factor-β1 (TGF-β1)/Smad signaling molecules in rats.METHODS:A single dose of bleomycin was intratracheal injected into the SD rats to induce pulmonary fibrosis .After injection of bleo-mycin, the rats were randomly divided into low-, medium-and high-dose rhein treatment groups and model group .The rats that were instilled with normal saline intratracheally served as control group .After the treatment for 28 d, the pulmonary pathologic changes were observed under microscope with hematoxylin-eosin staining .The lung coefficient and hydroxypro-line content were also measured .The expression of miR-21 and the mRNA levels of TGF-β1 and Smad7 in the lung tissues were detected by real-time PCR.The protein levels of TGF-β1 and Smad7 were determined by Western blot .RESULTS:Rhein significantly attenuated the experimental alveolitis , pulmonary fibrosis , lung coefficient and hydroxyproline contents in the rats.Rhein obviously decreased the expression of miR-21,and the mRNA and protein levels of TGF-β1, but signifi-cantly increased the mRNA and protein levels of Smad 7 in the lung tissues .CONCLUSION: Rhein effectively prevents bleomycin-induced pulmonary fibrosis by inhibiting the expression of miR-21 and promoting the expression of Smad 7, thus regulating the TGF/Smad signaling pathway to decrease extracellular matrix deposition .
5.Effect of up-regulation of miR-132 on expressions of angiopoietin-1/endothelium-specific tyrosine kinase receptor 2 in focal cerebral ischemia reperfusion injury of rats
Peng CHEN ; Chuanlei CHEN ; Aiwu HOU ; Qian GAO ; Shijuan ZHANG ; Xibo SUN ; Bingxuan LI ; Jianyi NIU
Chinese Journal of Neuromedicine 2017;16(8):772-776
Objective To investigate the effect of up-regulation of miR-132 on expressions of angiopoietin-1 (Ang-1)/endothelium-specific tyrosine kinase receptor 2 (Tie2) in focal cerebral ischemia reperfusion injury of rats.Methods Forty adult healthy SD rats were randomly divided into sham-operated group,cerebral ischemia group,miR-132 mimic group and negative control group (n=10).The models of middle cerebral artery occlusion in the later three groups were established by using modified Longa suture method.Rats in the miR-132 mimic group and negative control group were injected miR-132 mimic 15 μg and negative control 15 μg via paracele.Rats in each group were sacrificed 24 h after ischemia,and the brain tissues were collected;the total infarct volumes were calculated by TTC staining.The mRNA expressions ofmiR-132,Ang-1 and Tie2 in ischemic cerebral cortex tissues were detected by real-time fluorescence quantitative PCR.The protein expressions of Ang-1,Tie2,CD31 and vessel endothelial growth factor (VEGF) in ischemic cerebral cortex tissues were detected by Western blotting.Results The total infarct volume in the miR-132 mimic group was (27.92±3.05) mm3,which was significantly smaller than that in the cerebral ischemia group and negative control group ([51.34±2.86] mm3 and [50.46±2.57] mm3,P<0.05).The relative mRNA expression levels of miR-132,A ng-1,Tie2,and the protein expression levels of Ang-1,Tie2,CD31,VEGF in the ischemic cerebral cortex tissues of the miR-132 mimic group were significantly higher than those in the negative control group,cerebral ischemia group and sham-operated group (P<0.05);and those in the negative control group and cerebral ischemia group were significantly higher than those in the sham-operated group (P<0.05).Conclusion Up-regulation ofmiR-132 expression could improve the ischemic states of ischemic stroke in rats,which might be related to Ang-1/Tie2 increased expressions to promote angiogenesis in ischemic brain tissues.