Preparation of Rat Gingival Mitochondria with an Improved Isolation Method

Table 3

Effects of leukotoxin and lipopolysaccharide on the respiratory control ratio (RCR) in mitochondrial fractions from rat gingival tissues.

	Concentration used	Respiratory control
	(  μg/mg Mt proteina)	ratio
Control (none)	0.0	1.82 ± 0.05
Leukotoxin	0.5	1.29 ± 0.05∗
	1.0	1.15 ± 0.03∗
	2.0	1.05 ± 0.03∗
Lipopolysaccharide
	5.0	1.26 ± 0.03∗
	10.0	1.13 ± 0.03∗
	25.0	1.10 ± 0.03∗

The respiratory control ratio (RCR) of the mitochondrial (Mt) fractions which was obtained from the conditions of 0.120 % (w/v) collagenase treatment and 50 sec homogenization. At a concentration range of 0.5–2.0   μg/mg Mt protein, leukotoxin was added to a 600   μL of 10 mM HEPES buffer (pH 7.4) containing 0.25 M sucrose and 10 mM KH₂PO₄ in the reaction container of an oxygen electrode apparatus, followed by the addition of 100   μg/mL rotenone and 0.6 mM succinic acid. Also, at a concentration range of 5.0–25.0   μg/mg Mt protein, lipopolysaccharide was added in the same manner. RCR was determined polarographically, as shown in Table 2.
aMt protein; mitochondrial protein.
∗Significantly different when compared with the control (none) at 𝑃<.05.