2.2.3. Assay of phagocytic index
The function of the macrophage cells was assessed via a carbon
clearance test performed on three mice from each group as
described by Liang et al. (2007). Mice were intravenously injected
via the tail vein with diluted India ink (4-fold diluted with saline)
at a dose of 0.05 ml/10 g body weight, timed immediately after the
ink injection. Blood specimens were collected at 2 min (t1) and
10 min (t2) from the retinal venous plexuses. A volume of 20 μl
blood was mixed with 2 ml 0.1% Na2CO3. The absorbance at
600 nm was measured on a UV–visible spectrophotometer with
0.1% Na2CO3 as the blank. The liver and the spleen were weighed
after the mice were sacrificed. The phagocytic index α was
calculated as follows:
α ¼ K1=3 A=ðB þ CÞ
K ¼ ðlg OD1lg OD2Þ=ðt2t1Þ
where A, B and C represent the body, liver and spleen weight,
respectively. OD1 is for t1, and OD2 for t2.
2.2.4. Assay of cytokines
IL-2, IL-4, IL-10 contents in serums were determined with
enzymatic method kits. All procedures were performed according
to manufacturer′s instructions.
2
2.2.3. Assay of phagocytic index
The function of the macrophage cells was assessed via a carbon
clearance test performed on three mice from each group as
described by Liang et al. (2007). Mice were intravenously injected
via the tail vein with diluted India ink (4-fold diluted with saline)
at a dose of 0.05 ml/10 g body weight, timed immediately after the
ink injection. Blood specimens were collected at 2 min (t1) and
10 min (t2) from the retinal venous plexuses. A volume of 20 μl
blood was mixed with 2 ml 0.1% Na2CO3. The absorbance at
600 nm was measured on a UV–visible spectrophotometer with
0.1% Na2CO3 as the blank. The liver and the spleen were weighed
after the mice were sacrificed. The phagocytic index α was
calculated as follows:
α ¼ K1=3 A=ðB þ CÞ
K ¼ ðlg OD1lg OD2Þ=ðt2t1Þ
where A, B and C represent the body, liver and spleen weight,
respectively. OD1 is for t1, and OD2 for t2.
2.2.4. Assay of cytokines
IL-2, IL-4, IL-10 contents in serums were determined with
enzymatic method kits. All procedures were performed according
to manufacturer′s instructions.
2
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