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Journal of Chinese Integrative Medicine ›› 2012, Vol. 10 ›› Issue (11): 1293-1302.doi: 10.3736/jcim20121114

• Original Experimental Research • Previous Articles     Next Articles

Ameliorative potentials of Syzygium jambolanum extract against arsenic-induced stress in L6 cells in vitro

Samadder Asmita,Das Jayeeta,Das Sreemanti,Biswas Raktim,Rahman Khuda-Bukhsh Anisur()   

  1. Cytogenetics and Molecular Biology Laboratory, Department of Zoology, University of Kalyani, Kalyani-741235, IndiaCytogenetics and Molecular Biology Laboratory, Department of Zoology, University of Kalyani, Kalyani-741235, India
  • Received:2012-04-14 Accepted:2012-07-06 Online:2012-11-20 Published:2018-11-15

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Objective: To determine the ameliorative potentials of Syzygium jambolanum (SJ) extract in L6 skeletal muscle cells in regard to arsenic-induced impairment of optimum glucose homeostasis and improper functioning of mitochondria.

Methods: Several study parameters like glucose level and mitochondrial functioning through indexes of pyruvate kinase, glucokinase and mitochondrial membrane potential were assessed. The expression of the relevant marker proteins and mRNAs like glucose transporter 4 (GLUT4), insulin receptor substrate 1 (IRS1), IRS2 and glucokinase for tracking down the signalling cascade were critically analyzed.

Results: Introduction of SJ extract could bring about positive modulation of various markers, by acting on GLUT4, thereby bringing about an attenuation of the arsenite-induced toxic conditions in L6 cells.

Conclusion: Syzygium jambolanum extract has considerable ameliorating potentials against arsenic-induced glucose imbalance and stress and has possibility of therapeutic use in the management of arsenic-induced toxicity including hyperglycemia.

Key words: Syzygium, plant extracts, arsenates, glucose transport proteins,facilitative, L6 skeletal muscle cells, in vitro

Figure 1

Thermal stability curve of Syzygium jambolanum extract"

Figure 2

Graphical representation of cell viability in different set of experimental cells Data are presented as mean±standard deviation, n=3; **P<0.01, vs normal control group; △△P<0.01, vs SA-treated group. SA: sodium arsenite; SJ: Syzygium jambolanum."

Figure 3

Morphological analysis of different sets of L6 cells before and after treatment (Light microscopy, ×200) A: Normal control; B: SA control; C: SA plus SJ (50 ng/mL); D: SA plus SJ (100 ng/mL). SA: sodium arsenite; SJ: Syzygium jambolanum."

Figure 4

Morphological analysis of nuclear condensation of different sets of L6 cells before and after treatment stained by DAPI (Fluorescence microscopy, ×200) A: Normal control; B: SA control; C: SA plus SJ (50 ng/mL); D: SA plus SJ (100 ng/mL). SA: sodium arsenite; SJ: Syzygium jambolanum; DAPI: 4′,6-diamidino-2-phenylindole."

Figure 5

Graphical representation of glucose level in different set of experimental cells Data are presented as mean±standard deviation, n=3; **P<0.01, vs normal control group; △△P<0.01, vs SA-treated group. SA: sodium arsenite; SJ: Syzygium jambolanum."

Figure 6

Graphical representation of pyruvate kinase activity and glutathione level in different sets of experimental cells A: Pyruvate kinase activity; B: Glutathione level. Data are presented as mean±standard deviation, n=3; **P<0.01, vs normal control group; △P<0.05; △△P<0.01, vs SA-treated group. SA: sodium arsenite; SJ: Syzygium jambolanum."

Figure 7

Analysis of mitochondrial membrane potential in L6 cells (Fluorescence microscopy, ×200) A: Normal control; B: SA control; C: SA plus SJ (50 ng/mL); D: SA plus SJ (100 ng/mL). SA: sodium arsenite; SJ: Syzygium jambolanum."

Figure 8

Flow cytometric analysis of mitochondrial membrane potential of L6 cells A: Normal control; B: SA control; C: SA plus SJ (50 ng/mL); D: SA plus SJ (100 ng/mL). SA: sodium arsenite; SJ: Syzygium jambolanum."

Figure 9

Immunofluorescence detection of GLUT4 distribution in different groups of L6 cells (Fluorescence microscopy, ×200) A: Normal control; B: SA control; C: SA plus SJ (50 ng/mL); D: SA plus SJ (100 ng/mL). SA: sodium arsenite; SJ: Syzygium jambolanum."

Figure 10

Densitometric expression levels of different mRNAs tested by RT-PCR Ln1: Normal control; Ln2: SA control; Ln3: SA plus SJ (50 ng/mL); Ln4: SA plus SJ (100 ng/mL). SA: sodium arsenite; SJ: Syzygium jambolanum; SA: sodium arsenite; RT-PCR: reverse transcriptase-polymerase chain reaction analysis; G3PDH: glyceraldehyde-3-phosphate-dehydrogenase; IRS: insulin receptor substrate; GLUT: glucose transporter."

Figure 11

Graphical representation of expression levels of different mRNAs tested by RT-PCR Data are presented as mean±standard deviation, n=3; **P<0.01, vs normal control group; △P<0.05; △△P<0.01, vs SA-treated group. SA: sodium arsenite; SJ: Syzygium jambolanum; RT-PCR: reverse transcriptase-polymerase chain reaction analysis; G3PDH: glyceraldehyde-3-phosphate-dehydrogenase; IRS: insulin receptor substrate; GLUT: glucose transporter."

Figure 12

Expression of different proteins tested by indirect ELISA technique Data are presented as mean±standard deviation, n=3; **P<0.01, vs normal control group; △P<0.05; △△P<0.01, vs SA-treated group. SA: sodium arsenite; SJ: Syzygium jambolanum; ELISA: enzyme-linked immunosorbent assay; IRS: insulin receptor substrate; GLUT: glucose transporter."

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