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Journal of Chinese Integrative Medicine ›› 2011, Vol. 9 ›› Issue (8): 894-900.doi: 10.3736/jcim20110812

• Original Experimental Research • Previous Articles     Next Articles

Effects of ethanol extract of Rhizome Pinelliae Preparata on intracellular pH value of human gastric adenocarcinoma cells

Ci-an Zhang, Feng Wu, Zhu-jun Mao, Zhen Wei, Yong-jin Li, Pin-kang Wei()   

  1. Department of Traditional Chinese Medicine, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China
  • Received:2011-04-23 Accepted:2011-05-11 Online:2011-08-20 Published:2011-08-15

Objective: To observe the effects of ethanol extract of Rhizome Pinelliae Preparata on the intracellular pH value of human gastric cancer SGC7901 cells.
Methods: After coculturing SGC7901 cells with ethanol extract of Rhizome Pinelliae Preparata (1, 0.5, 0.25 and 0.125 mg/mL), cell viability was evaluated by chromatometry with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) staining. Intracellular pH value of SGC7901 cells was measured in the monolayer by using the pH-sensitive fluorescent probe 2,7-bis-(2-carboxyethyl)-5-carboxyfluorescein-acetoxymethyl ester. The extracellular pH value of culture medium was measured by a pH211 Calibration Check Microprocessor pH Meter. Half-inhibitory concentration (IC50) of ethanol extract culture to SGC7901 cells was decided by the MTT method and expressions of vacuolar-H+-ATPase (V-ATPase) and Na+/H+ exchanger isoform 1 (NHE1) mRNAs were examined by the method of fluorescence quantitative-polymerase chain reaction after 72 h of drug treatment.
Results: Ethanol extract of Rhizome Pinelliae Preparata at different concentrations significantly inhibited the proliferation of SGC7901 cells, lowered the intracellular pH values and heightened the extracellular pH values. The IC50 of 72 h culture was 0.5 mg/mL and it inhibited the expressions of V-ATPase and NHE1 mRNAs.
Conclusion: Ethanol extract of Rhizome Pinelliae Preparata can lower down the intracellular pH value of SGC7901 cells. The mechanism may be related to inhibiting the expressions of V-ATPase and NHE1 mRNAs.

Key words: drugs,Chinese herbal, Rhizome Pinelliae Preparata, stomach neoplasms, intracellular pH value, sodium-hydrogen antiporter, vacuolar proton-translocating ATPases, cell line, tumor

Table 1

Effects of ethanol extract of Rhizome Pinelliae Preparata on proliferation of SGC7901 cells ($\bar{x}$±s)"

Group n Optical density value
24 h 48 h 72 h
Control 5 0.317±0.013 0.563±0.071 0.824±0.063
Ethanol extract 0.125 mg/mL 5 0.305±0.037 0.500±0.077 0.689±0.102**
Ethanol extract 0.25 mg/mL 5 0.292±0.025* 0.473±0.087* 0.614±0.058**
Ethanol extract 0.5 mg/mL 5 0.286±0.026* 0.373±0.095**△△ 0.485±0.030**△△
Ethanol extract 1 mg/mL 5 0.246±0.021**△△ 0.263±0.046**△△ 0.203±0.061**△△

Figure 1

Inhibitory effects of ethanol extract of Rhizome Pinelliae Preparata on SGC7901 cells"

Figure 2

Standard curve of intracellular pH values of SGC7901 cells BCECF: 2,7-bis-(2-carboxyethyl)-5-carboxyfluorescein."

Table 2

Effects of ethanol extract of Rhizome Pinelliae Preparata on intracellular pH values of SGC7901 cells ($\bar{x}$±s)"

Group n Intracellular pH
0 h 24 h 48 h 72 h
Control 5 7.51±0.03 7.50±0.04 7.51±0.05 7.50±0.06
Ethanol extract 0.125 mg/mL 5 7.51±0.03 7.50±0.04 7.49±0.03 7.47±0.03
Ethanol extract 0.25 mg/mL 5 7.51±0.03 7.49±0.03 7.46±0.03** 7.44±0.02**
Ethanol extract 0.5 mg/mL 5 7.51±0.03 7.47±0.03* 7.43±0.02**△△ 7.40±0.04**△△
Ethanol extract 1 mg/mL 5 7.51±0.03 7.40±0.02**△△ 7.35±0.01**△△ 7.31±0.03**△△

Table 3

Effects of ethanol extract of Rhizome Pinelliae Preparata on extracellular pH values of SGC7901 cells ($\bar{x}$±s)"

Group n Extracellular pH
0 h 24 h 48 h 72 h
Control 5 7.38±0.02 7.17±0.02 7.06±0.04 7.01±0.06
Ethanol extract 0.125 mg/mL 5 7.38±0.02 7.18±0.02 7.08±0.05 7.03±0.03
Ethanol extract 0.25 mg/mL 5 7.38±0.02 7.19±0.04 7.10±0.02* 7.06±0.04*
Ethanol extract 0.5 mg/mL 5 7.38±0.02 7.26±0.03**△△ 7.21±0.05**△△ 7.18±0.04**△△
Ethanol extract 1 mg/mL 5 7.38±0.02 7.32±0.03**△△ 7.28±0.04**△△ 7.25±0.04**△△

Figure 3

Effects of ethanol extract of Rhizome Pinelliae Preparata on pH gradient (extracellular — intracellular) in SGC7901 cells"

Figure 4

PCR amplification kinetics GAPDH: glyceraldehyde-3-phosphate dehydrogenase; V-ATPase: vacuolar-H+-ATPase; NHE1: Na+/H+ exchanger isoform 1; PCR: polymerase chain reaction."

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