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Journal of Chinese Integrative Medicine ›› 2010, Vol. 8 ›› Issue (7): 662-669.doi: 10.3736/jcim20100710

• Original Experimental Research • Previous Articles     Next Articles

Wuling Capsule promotes hippocampal neurogenesis by improving expression of connexin 43 in rats exposed to chronic unpredictable mild stress

Li De-qiang,Li Xu-juan,Duan Jin-feng,Cai Wei()   

  1. Department of Neurology, the First Affiliated Hospital, College of Medicine,Zhejiang University, Hangzhou 310003, Zhejiang Province, China
  • Received:2009-12-10 Online:2010-07-20 Published:2018-10-20
  • Contact: Cai Wei

Objective: To investigate the effects of Wuling Capsule, a compound traditional Chinese herbal medicine, on hippocampal neurogenesis by examining the expressions of brain-derived neurotrophic factor (BDNF) and connexin 43 (Cx43) in rats with depression induced by chronic unpredictable mild stress (CMS), and thereby to explore its antidepressant mechanism.
Methods: Forty-five adult male Sprague-Dawley rats were randomly divided into three groups: control group (n=15), untreated group (n=15) and Wuling group (n=15). All rats except those in the control group were subjected to 3-week CMS to induce depression. At the same time Wuling Capsule was daily added to the diet of the rats in the Wuling group at a dose of 100 mg/kg body weight for 21 days. The degree of depression was determined by sucrose preference test. BDNF expression and neurogenesis were tested by using immunohistochemical staining with BDNF and 5-bromodeoxyuridine (BrdU) antibodies; and the mRNA and protein expression levels of Cx43 in hippocampus were examined by semi-quantitative reverse transcription-polymerase chain reaction and Western blotting.
Results: The numbers of BDNF-positive neurons and BrdU-positive particles in dentate gyrus (DG) were significantly decreased in CMS rats as compared with the normal rats, and the same changes were found in Cx43 mRNA and protein expressions. After Wuling Capsule treatment, the depressed behaviors were improved. Moreover, the reduced expression levels of Cx43 mRNA and protein and fewer newborn neurons induced by CMS were recovered to the normal levels. However, BDNF-positive cells remained low in DG.
Conclusion: Wuling Capsule can improve the low hippocampal neurogenesis in rats subjected to CMS and the antidepressant effects are related to enhancing the Cx43 expression but not through BDNF mediation.

Key words: chronic unpredictable mild stress, brain-derived neurotrophic factor, 5-bromodeoxyuridine, connexin, rats

Figure 1

Sucrose preference of rats in three groups"

Figure 2

Expression of BDNF and number of BDNF-positive cells in DG of hippocampus of rats in three groups A: Photograph of BDNF expression detected by SABC immunohistochemical staining and the counting position (Light microscopy, ×200) in DG of hippocampus. B: Analysis of BDNF expression by SABC immunohistochemical staining (Light microscopy, ×400) in DG of hippocampus showing the morphous of the BDNF-positive cells among three groups and data were represented as x±s. n=15. 1: Control group; 2: Untreated group; 3: Wuling group. Bars in the photograph represent 50 μm. *P<0.05, vs control group."

Figure 3

Expression of BrdU and number of new-born neurons in DG of hippocampus of rats in three groups 1: Control group; 2: Untreated group; 3: Wuling group. Photographs (A, B, C) show BrdU expression in DG of the hippocampus detected by SABC immunohistochemical staining (Light microscopy, ×100 and ×400). Bars in the photographs represent 250 μm. Number of new-born neurons (D) was represented as x±s. n=15. *P<0.05, vs control group; △P<0.05, vs untreated group."

Figure 4

Expression of Cx43 mRNA in hippocampus in three groups The left graph is 2% agarose gel images showing specific Cx43 and GAPDH mRNA expressions, and the right bar graph shows the results of relative intensity of Cx43 mRNA expression to GAPDH mRNA. Data were represented as ?x±s. n=15. *P<0.05, vs control group; △P<0.05, vs untreated group."

Figure 5

Expression of Cx43 protein in hippocampus in three groups The left graph is Western blotting images, and the right bar graph shows the ratio of Cx43 protein to β-actin protein. Data were represented as ?x±s. n=15. *P<0.05, vs control group; △P<0.05, vs untreated group."

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