Skip to main page content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2012 May;33(5):578-87.
doi: 10.1038/aps.2012.3. Epub 2012 Mar 26.

Asiatic Acid, a Pentacyclic Triterpene in Centella Asiatica, Attenuates Glutamate-Induced Cognitive Deficits in Mice and Apoptosis in SH-SY5Y Cells

Affiliations
Free PMC article

Asiatic Acid, a Pentacyclic Triterpene in Centella Asiatica, Attenuates Glutamate-Induced Cognitive Deficits in Mice and Apoptosis in SH-SY5Y Cells

Min-fang Xu et al. Acta Pharmacol Sin. .
Free PMC article

Abstract

Aim: To investigate whether asiatic acid (AA), a pentacyclic triterpene in Centella asiatica, exerted neuroprotective effects in vitro and in vivo, and to determine the underlying mechanisms.

Methods: Human neuroblastoma SH-SY5Y cells were used for in vitro study. Cell viability was determined with the MTT assay. Hoechst 33342 staining and flow cytometry were used to examine the apoptosis. The mitochondrial membrane potential (MMP) and reactive oxygen species (ROS) were measured using fluorescent dye. PGC-1α and Sirt1 levels were examined using Western blotting. Neonatal mice were given monosodium glutamate (2.5 mg/g) subcutaneously at the neck from postnatal day (PD) 7 to 13, and orally administered with AA on PD 14 daily for 30 d. The learning and memory of the mice were evaluated with the Morris water maze test. HE staining was used to analyze the pyramidal layer structure in the CA1 and CA3 regions.

Results: Pretreatment of SH-SY5Y cells with AA (0.1-100 nmol/L) attenuated toxicity induced by 10 mmol/L glutamate in a concentration-dependent manner. AA 10 nmol/L significantly decreased apoptotic cell death and reduced reactive oxygen species (ROS), stabilized the mitochondrial membrane potential (MMP), and promoted the expression of PGC-1α and Sirt1. In the mice models, oral administration of AA (100 mg/kg) significantly attenuated cognitive deficits in the Morris water maze test, and restored lipid peroxidation and glutathione and the activity of SOD in the hippocampus and cortex to the control levels. AA (50 and 100 mg/kg) also attenuated neuronal damage of the pyramidal layer in the CA1 and CA3 regions.

Conclusion: AA attenuates glutamate-induced cognitive deficits of mice and protects SH-SY5Y cells against glutamate-induced apoptosis in vitro.

Figures

Figure 1
Figure 1
Inhibition by AA of Glu-induced cell death. (A) Cell viability was determined using MTT assay. Cells were exposed to 2–10 mmol/L Glu for 24 h without AA pretreatment. (B) Cells were preincubated with different concentrations of AA for 24 h, and then exposed to 10 mmol/L Glu for 24 h. bP<0.05 vs control. eP<0.05 vs Glu. All data are expressed as mean±SD of 5 replicate values in 3 separate experiments.
Figure 2
Figure 2
AA pretreatment significantly suppressed Glu-induced cell apoptosis. (A) AA pretreatment significantly decreased Glu-induced nuclear condensation. Apoptosis was determined morphologically by staining with Hoechst 33342 (original magnification 400×). AA10, 10 nmol/L AA. (B) Flow cytometric analyses with Annexin V-FITC and PI labels of cultured SH-SY5Y cells. (C) Percentage of apoptotic cells in total cells (n=3). bP<0.05 vs control. eP<0.05 vs Glu.
Figure 3
Figure 3
AA reduced ROS generation and restored MMP following Glu stimulus. (A) Cells were exposed to 10 mmol/L Glu with or without 10 nmol/L AA pretreatment. Representative fluorescence photographs (100×). AA10: 10 nmol/L AA. (B) Levels of ROS were analysed by FACS. Representative flow cytometry graph showing the DCF staining (n=3). Red peak: control; green peak: 10 mmol/L Glu; blue peak: 10 nmol/L AA+10 mmol/L Glu. (C) MMP of cells with 24 h Glu stimulus in the absence or presence of AA, determined by fluorescence spectrometry. (D) Representative fluorescence photographs of cellular mitochondrial membrane potential (400×). Mean±SD (n=3) are shown. bP<0.05 vs control. eP<0.05 vs Glu.
Figure 4
Figure 4
Effects of AA on Glu-stimulated expressions of Sirt1 and PGC-1α. Sirt1 and PGC-1α expressions in cells pretreated with AA (0.1–10 nmol/L). Mean±SD. n=3. bP<0.05 vs Glu group.
Figure 5
Figure 5
Effects of MSG and treatment with AA on the performance of spatial memory acquisition. (A) Latencies of mice during the 4 d test. (B) Times on d 5 that mice spent in the target quadrant with the platform removed. Mean±SEM. n=10. bP<0.05 vs control. eP<0.05 vs MSG.
Figure 6
Figure 6
Effect of AA treatment on CA1 and CA3 neuron densities in hippocampus. Representative pictures obtained by HE staining (400×). When compared with the MSG-treated group, CA1 and CA3 neurons appear well preserved in AA-treated mice. MSG, 2.5 mg/g MSG; AA50, 50 mg/kg AA; AA100: 100 mg/kg AA.

Similar articles

See all similar articles

Cited by 32 articles

See all "Cited by" articles

Publication types

MeSH terms

Feedback