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©2005 Baishideng Publishing Group Inc.
World J Gastroenterol. Feb 14, 2005; 11(6): 823-830
Published online Feb 14, 2005. doi: 10.3748/wjg.v11.i6.823
Published online Feb 14, 2005. doi: 10.3748/wjg.v11.i6.823
Table 1 Red cell membrane preparation procedure.
Whole blood | |
↓ | Centrifuged at 3000 r/min for 10 min, rinsed twice with 1:3 saline, and suspended in 1:1 saline |
Red cell suspension | |
↓ | 5p 8.4 phosphoric buffer solution (1:30) was added at 4 °C and hemolysis was allowed to proceed for 1 h. |
Hemolyzed solution of red cells | |
↓ | Centrifuged at 2000 r/min at 4 °C for 40 min; the supernatant was discarded; the residue was rinsed 3 times and then suspended in 5p8.4 phosphoric acid buffer solution. |
Membrane suspension |
Table 2 Steps for measuring the activity of the sialic acid content on the surface of the red cell membrane.
Reagent | Blank | 1 | 2 | 3 | 4 | 5 | 6 | 7 |
N-acetyl neuraminic acid Standard solution (mL) | 0 | 0.05 | 0.1 | 0.2 | 0.3 | 0.4 | 0.5 | 0.8 |
N-acetyl neuraminic acid (mg) | 0 | 5 | 10 | 20 | 30 | 40 | 50 | 80 |
Distilled water (mL) | 1 | 0.95 | 0.9 | 0.8 | 0.7 | 0.6 | 0.5 | 0.2 |
Bialsche reagent (mL) | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 1 |
Boil for 12 min in water bath, then cool for 3 min with ice water | ||||||||
Pentanol (mL) | 5 | 5 | 5 | 5 | 5 | 5 | 5 | 5 |
Shaken thoroughly and then centrifuged for 10 min at 1 000 r/min; took the pentanol phase for colorimetric analysis at a wavelength 569 nm |
Table 3 Steps for measuring the activity of the oxidoreductase NADH-cell pigment C.
Reagent (mL) | No saponin added (-S) | Saponin added (+S) | ||
Blank | Sample | Blank | Sample | |
0.6 mg protein/mL Membrane suspension | 0.6 | 0.6 | 0.6 | 0.6 |
PBS (5p8.4) | 2.4 | 2.2 | 2.2 | 2 |
0.1% saponin | / | / | 0.2 | 0.2 |
5 mmol/L K3Fe(CN)6 | / | 0.1 | / | 0.1 |
Let the solution stand for 10 min, added NADH, and recorded the result after 1 min 6 mmol/L NADH | / | 0.1 | / | 0.1 |
Table 4 Effect of Haimiding on the immunological adhesiveness of red cells of FC mice to tumor cells (mean±SD).
Group | Number of animals (n) | Route of administration | Dosage (mg/kg) | Rate of wreaths (%) |
Control | 10 | Ip | 36.36±5.42 | |
Saline | 10 | Ip | Equal volume | 10.46±3.55 |
5-Fu | 10 | Ip | 25 | 8.41±2.56 |
Haimiding | 10 | Ip | 50 | 28.96±4.41b |
Table 5 Effect of Haimiding on the immunological adhesiveness of red cells of H22 mice to tumor cells mean±SD.
Group | Dosage (mg/kg) | Route of administration | No. of animals (n) | Rate of wreaths (%) |
Control | Ip | 10 | 36.36±5.42 | |
Saline | Equal volume | Ip | 10 | 5.10±2.28 |
5-Fu | 25 | Ip | 10 | 5.96±3.15 |
Haimiding | 50 | Ip | 10 | 25.68±4.13b |
Table 6 Effect of Haimiding on the fluidity of intact red cell membrane of FC mice.
Group | Number ofanimals (n) | Route ofadministration | Dosage (mg/kg) | Fluorescencepolarization (P) | Micro viscosity, h(P) | Membrane lipidfluidity (LFU) |
Control | 10 | Equal volume | 0.2006±0.0728 | 1.7557±0.9616 | 7.4472±2.3273 | |
Saline | 10 | Ip | 25 | 0.2438±0.0122 | 1.9458±0.2367 | 4.3134±1.4378 |
5-Fu | 10 | Ip | 100 | 0.2169±0.0269 | 1.8183±0.3799 | 6.0234±1.9430a |
Haimiding | 10 | Ip | 50 | 0.215±0.0242b | 1.796±0.1414a | 6.1913±1.9348a |
Haimiding | 10 | Ip | 25 | 0.210±0.0137b | 1.701±0.2063a | 6.4966±1.9687b |
Haimiding | 10 | Ip | 0.2203±0.0338 | 1.7330±0.4404 | 5.7670±1.6962a |
Table 7 Effect of Haimiding on the fluidity of intact red cell membrane of H22 mice.
Group | Number ofanimals (n) | Route ofadministration | Dosage (mg/kg) | Fluorescencepolarization (P) | Micro viscosity, h(P) | Membrane lipidfluidity (LFU) |
Control | 10 | 0.2006±0.0728 | 1.7557±0.9616 | 7.4478±2.3273 | ||
Saline | 10 | Ip | Equal volume | 0.2430±0.03017 | 2.1378±0.6186 | 4.3559±1.4051 |
5Fu | 10 | Ip | 25 | 0.2275±0.0182 | 1.9760±0.6738 | 5.2606±1.5472 |
Haimiding | 10 | Ip | 100 | 0.2331±0.0211 | 2.0819±0.1772 | 4.9153±1.5360 |
Haimiding | 10 | Ip | 50 | 1.2775±0.4164b | 11.0370±3.4489b | 0.1770±0.0567b |
Haimiding | 10 | Ip | 25 | 0.2111±0.0223a | 1.8176±0.2106 | 6.4776±2.0895a |
Table 8 Effect of Haimiding on the membrane protein content in the red cells of FC mice.
Group | Number of animals (n) | Route of administration | Dosage (mg/kg) | Membrane protein content % |
Control | 10 | Ip | Equal volume | 0.281±0.051 |
Saline | 10 | Ip | 25 | 0.332±0.094 |
5-Fu | 10 | Ip | 100 | 0.278±0.041 |
Haimiding | 10 | Ip | 50 | 0.241±0.037a |
Haimiding | 10 | Ip | 25 | 0.187±0.042b |
Haimiding | 10 | Ip | 0.296±0.040 |
Table 9 Effect of Haimiding on the membrane protein content in the red cells of H22 mice.
Group | Number of animals (n) | Route of administration | Dosage (mg/kg) | Membrane protein content % |
Control | 10 | Equal volume | 0.281±0.051 | |
Saline | 10 | Ip | 25 | 0.298±0.088 |
5-Fu | 10 | Ip | 100 | 0.219±0.089 |
Haimiding | 10 | Ip | 50 | 0.237±0.050 |
Haimiding | 10 | Ip | 25 | 0.143±0.064b |
Haimiding | 10 | Ip | 0.1890.094a |
Table 10 Effect of Haimiding on the sialic acid content on the surface of the red cell membrane of FC mice.
Group | Number of animals (n) | Route of administration | Dosage (mg/kg) | A |
Control | 10 | Ip | 0.183±0.068 | |
Saline | 10 | Ip | Equal volume | 0.087±0.026 |
5-Fu | 10 | Ip | 25 | 0.065±0.030 |
Haimiding | 10 | Ip | 100 | 0.139±0.049b |
Haimiding | 10 | Ip | 50 | 0.148±0.032b |
Haimiding | 10 | Ip | 25 | 0.095±0.028 |
Table 11 Effect of Haimiding on the sialic acid content on the surface of the red cell membrane of H22 mice.
Group | Number of animals (n) | Route of administration | Dosage (mg/kg) | A |
Control | 10 | Ip | 0.183±0.068 | |
Saline | 10 | Ip | Equal volume | 0.062±0.027 |
5-Fu | 10 | Ip | 25 | 0.064±0.034 |
Haimiding | 10 | Ip | 100 | 0.119±0.040b |
Haimiding | 10 | Ip | 50 | 0.125±0.044b |
Haimiding | 10 | Ip | 25 | 0.081±0.023 |
Table 12 Effect of Haimiding on the sealability of the red cell membrane of FC mice.
Group | Number of animals (n) | Route of administration | Dosage (mg/kg) | Sealability (%) |
Control | 10 | Ip | 70.6±4.05 | |
Saline | 10 | Ip | Equal volume | 54.6±2.21 |
5-Fu | 10 | Ip | 25 | 52.0±3.39 |
Haimiding | 10 | Ip | 100 | 63.5±3.11b |
Haimiding | 10 | Ip | 50 | 65.9±2.60b |
Haimiding | 10 | Ip | 25 | 58.4±2.05b |
Table 13 Effect of Haimiding on the sealability of the red cell membrane of H22 mice.
Group | Number of animals (n) | Route of administration | Dosage (mg/kg) | Sealability (%) |
Control | 10 | Ip | 70.2±2.23 | |
Saline | 10 | Ip | Equal volume | 53.7±2.30 |
5-Fu | 10 | Ip | 25 | 54.2±3.27 |
Haimiding | 10 | Ip | 100 | 62.2±2.71b |
Haimiding | 10 | Ip | 50 | 61.4±3.44b |
Haimiding | 10 | Ip | 25 | 57.6±2.54b |
- Citation: Ji YB, Gao SY, Cheng WP. Effect of Haimiding on the functioning of red cell membrane of FC and H22 tumor-bearing mice. World J Gastroenterol 2005; 11(6): 823-830
- URL: https://www.wjgnet.com/1007-9327/full/v11/i6/823.htm
- DOI: https://dx.doi.org/10.3748/wjg.v11.i6.823