1.Study on synergistic antidepressant effect of 3,6-disinapoyl sucrose combined with tenuifoliside A and its possible mechanism
Runqing ZHAO ; Yuan HU ; Muhan LI ; Jing ZHANG ; Xiao TAN ; Ping LIU
Chinese Pharmacological Bulletin 2016;32(5):716-721,722
Aim To study the synergistic anti-depres-sion effect of 3 , 6-disinapoyl sucrose ( DISS ) and tenuifoliside A ( TFSA ) from Radix Polygalae and the preliminary mechanism . Methods Using the classical behavioral despair and depression model of mouse tail suspension test, 120 mice were divided into control group, positive group, DISS 5 mg·kg-1 group,DISS 10 mg·kg-1 group,TFSA 5 mg·kg-1 group,TFSA 10 mg· kg-1 group, DISS 5 mg · kg-1 +TFSA 5 mg · kg-1 group,DISS 5 mg·kg-1 +TFSA 10 mg·kg-1 group,DISS 10 mg·kg-1 +TFSA 5 mg·kg-1 group and DISS 10 mg · kg-1 +TFSA 10 mg · kg-1 group randomly. They were given intragastric injection for 7 days continuously, to observe the effect of DISS and TFSA monomer and its combination on the time of mouse tail suspension. Expression of BDNF in the hip-pocampus of mice was detected by immunohistochemis-try. The expressions of CREB, pCREB, CRTC1 and BDNF in the hippocampus of mice were detected by Western blot method. Results The administration of DISS and TFSA could shorten the immobility time of mice subjected to the tail. DISS ( 10 mg · kg-1 ) and TFSA( 10 mg · kg-1 ) group were significantly lower than single dose drug group(P<0. 05). DISS and TF-SA and the combination groups could increase the ex-pression of BDNF in hippocampus and cortex by immu-nohistochemistry(P <0. 05). At the same time, the contents of CREB, CRTC1, pCREB and BDNF protein in the hippocampus were increased by DISS and TF-SA, and the combination group was significantly higher than the single drug group ( P<0. 05 ) . Conclusion The administration of DISS and TFSA are used to acti-vate CREB transcription factor CRTC1 , and activate the phosphorylation of CREB in the hippocampus, and then increase the expression of BDNF in the hippocam-pus and plays a synergistic antidepressant effect.
2.Discovery and functional verification of endogenous glucanases for scleroglucan hydrolysis in Sclerotium rolfsii.
Weizhu ZENG ; Runqing TAN ; Jingwen ZHOU
Chinese Journal of Biotechnology 2021;37(1):207-217
Scleroglucan is a high-molecular water-soluble microbial exopolysaccharide and mainly applied in the fields of petroleum, food, medicine and cosmetics. The high molecular weight of scleroglucan produced by microbial fermentation leads to low solubility, high viscosity and poor dispersibility, thus bringing a series of difficulties to extraction, preservation and application. It is important to explore suitable degradation method to adjust the molecular weight of scleroglucan for expanding its industrial application. Taking Sclerotium rolfsii WSH-G01 as a model strain, in which functional annotations of the glucanase genes were conducted by whole genome sequencing. Based on design of culture system for culture system for differential expression of β-glucanase, endogenous β-glucanase genes in S. rolfsii WSH-G01 were excavated by transcriptomics analysis. Functions of these potential hydrolases were further verified. Finally, 14 potential endogenous hydrolase genes were obtained from S. rolfsii. After heterologous overexpression in Pichia pastoris, 10 soluble enzymes were obtained and 5 of them had the activity of laminarin hydrolysis by SDS-PAGE and enzyme activity analysis. Further investigation of the 5 endogenous hydrolases on scleroglucan degradation showed that enzyme GME9860 has positive hydrolysis effect. The obtained results provide references not only for obtaining low and medium molecular weight of scleroglucan with enzymatic hydrolysis, but also for producing different molecular weight of scleroglucan during S. rolfsii fermentation process with metabolic engineering.
Basidiomycota/genetics*
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Glucans
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Hydrolysis
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Saccharomycetales
3.CCL2/CCR2 signaling activation contributes to tooth movement pain
Zhi YANG ; Wei LUO ; Runqing FU ; Yu TAN ; Lingjun YUAN ; Bing FANG
Chinese Journal of Stomatology 2014;49(8):500-505
Objective To test the hypothesis that the CCL2/CCR2 signaling pathway plays an important role in pain induced by experimental tooth movement.Methods Male Sprague-Dawley rats weighing between 200 and 300 g were used in this study.Expression of CCL2/CCR2 in the trigeminal ganglion(TG) was determined by Western blotting 0 h,4 h,1 d,3 d,5 d,7 d after tooth movement.Localization of the CCL2 was revealed by immunohistochemistry.Changes in body weight,nocifensive behaviors,and the effects of CCL2/CCR2 antagonists on these changes in pain behaviors were evaluated.Exogenous CCL2 was injected into periodontal tissues and added to TG neurons in culture and the resulting c-fos expression and pain responses were detected.In addition,the expression and cellular localization of CCL2 in the medullary dorsal horn (MDH) was determined by immunohistochemistry 3 d and 14 d after tooth movement.Results Experimental tooth movement led to a statistically significant increase in CCL2/CCR2 expression at the protein level from day 3 to 7 after application of force initiating tooth movement.When compared with control group(1.000± 0.000),CCL2 increased to (2.620 ± 0.128),(3.300±0.197) and (1.740±1.290) at day 3,5 and 7 respectively,which were statistically significant (P<0.05).CCR2 expression levels were (1.636±0.061) and (1.766±0.126) compared with that in control group (1.000±0.000) at day 3 and 5 respectively with statistical significance (P<0.05).Both of them peaked on day 5 (3.3 and 1.8 time compared to control group).Application of recombinant CCL2 led to the up-regulation of c-fos expression in vivo and in vitro,and triggered a corresponding nocifensive behavior in rats.The magnitude of the nocifensive behavior could be reduced by a CCR2 antagonist,and by CCL2 neutralizing antibody.Furthermore,we found a significant increase in the expression of CCL2,corresponding well to the upregulation of the time spent on nocifensive behaviors after ETM.In addition,CCL2 was up-regulated in TG neurons and astrocytes in Vc.Conclusions The CCL2/CCR2 axis was modulated by experimental tooth movement and involved in the development of tooth movement pain,and thus palyed an important role in orthodontic pain mechanism.