SCLM, total saponins extracted from Chaihu-jia-longgu-muli-tang, reduces chronic mild stress-induced apoptosis in the hippocampus in mice
Authors: Yadong Liu
Shiping Ma
Rong Qu
Journal: Pharmaceutical Biology (2010)
Discipline: PHARMACOLOGY & TOXICOLOGY
Keywords: stress; depression; hippocampus; apoptosis; neuroprotective
Affiliations: Department of Pharmacology of Chinese Materia Medica, China Pharmaceutical University, Nanjing, People's Republic of China.

Abstract:  Increasing evidence demonstrates that stress or depression can lead to atrophy and cell loss in the hippocampus. In contrast, antidepressant treatment significantly reduces apoptosis in the dentate granule cell layer and subgranular zone in animal models of depression. In the present study, we investigated the neuroprotective action of SCLM, the total saponins extracted from Chaihu-jia-longgu-muli-tang, a traditional Chinese medicinal formula which was prescribed 1000 years ago, in the reduction of apoptosis in hippocampal neurons using an experimental chronic mild stress (CMS) model. Mice were subjected to the CMS procedure for a period of 21 consecutive days. SCLM (100 mg/kg, p.o.) or fluoxetine (20 mg/ kg, p.o.) was administered during the stress periods. CMS mice showed a decreased sucrose intake over 21 days, and an increase in the number of TUNEL-positive neurons as well as up-regulation of the apoptotic-related factors, such as Bax and caspase-3 in the hippocampus, compared with control mice. On the other hand, the administration of SCLM (100 mg/kg) and fluoxetine (20 mg/kg) reversed these effects induced by CMS, showing a significant increase of sucrose intake and a dramatic reduction of TUNEL-positive neurons and decreased expression of Bax and caspase-3 proteins. The present results suggest that SCLM possesses a significant antidepressant-like property, and this effect may be through protection against stress-induced neuronal apoptosis by affecting the expression of Bax and caspase-3 proteins in the hippocampus. These findings provide important information that the anti-apoptotic effect of herbal medicine therapy may be beneficial for the treatment of depression. 

Retracted: No

PubPeer comments:
  Figure 2. (a) SCLM inhibited up-regulation of Bax in the hippocampus of stressed mice. (A) Control; (B) stressed mice: numerous Bax-positive neurons could be observed, dark brown (arrow); (C) fluoxetine-treated (20mg kg) stressed mice: Bax-positive neurons were significantly decreased; (D) SCLM-treated (100mg kg) stressed mice: Bax-positive neurons were significantly reduced. C appears to be an enlargement of A, though with additional cells becoming visible or changing color. ![file] D and (B display a similar relationship. Here rescaled for ease of comparison. ![file] , 
  Figure 1. (a) SCLM inhibited levels of TUNEL-positive cells in the hippocampus of stressed mice. (A) Control; (B) stressed mice: numerous TUNEL-positive neurons could be observed, dark brown (arrows); (C) fluoxetine-treated (20mg kg) stressed mice: TUNEL-positive neurons were significantly decreased (arrows) . I have marked a few of the internal similarities. ![file] , 
 Fig 3 (a)  SCLM inhibited up-regulation of caspase-3 in the hippocampus of stressed mice. (A) Control; (B) stressed mice: numerous neurons with positive staining of caspase-3 could be observed, dark brown (arrow); (C) fluoxetine-treated (20mg kg) stressed mice: caspase-3-positive neurons were significantly reduced; (D) SCLMtreated (100mg kg) stressed mice: caspase-3-positive neurons were significantly decreased . Panels C and D are the same, in an unfortunate error. I am more interested in the marked similarities within B. ![file] A and C reveal a surprising overlap (after some rescaling). ![file] , 
 Dear Hoya, Thank you for performing query on our manuscript. We are really sorry for the low-quality representative pictures from immunohistochemical staining. In order to obtain optimal vision field and get clear photos in the microscope, we used different magnifications for each mouse brain slice. When we zoomed and cropped the original images to collectively present the representative pictures, we accidentally inserted the wrong images into the combined four groups, leading to some overlap and similar pictures between and within groups. Itu2019s our fault for not critical checking these important images before submission and during revision and proof stages. Itu2019s also our responsibility to correct this mistake by providing high-quality images after revisiting the raw data. Meanwhile, we will send an erratum request to the editor of Pharmaceutical Biology shortly and resubmit the correct images immediately. We apologize for any negative impact caused by this unintended error. We believe our update data will give you additional confidence in the results we presented in this paper. Regards, Yadong , 
 #4 Thank you for your reply. It will be interesting to see how the journal views the repetitive features clearly identified in #2 and #3. It is difficult to understand how these concerns can be addressed with higher resolution images. ,