Longxuetongluo Capsule Improves Erythrocyte Function against Lipid Peroxidation and Abnormal Hemorheological Parameters in High Fat Diet-Induced ApoE-/- Mice

Abstract

Chinese dragon's blood, the red resin of Dracaena cochinchinensis, one of the renowned traditional medicines, has been used to facilitate blood circulation and disperse blood stasis for thousands of years. Phenolic compounds are considered to be responsible for its main biological activities. In this study, total phenolic compounds of Chinese dragon's blood were made into capsule (Longxuetongluo Capsule, LTC) and their effects on the abnormal hemorheological properties were examined by high fat diet (HFD) induced ApoE-/- mice. Compared to the model group, LTC recovered the abnormal hemorheological parameters in HFD-induced ApoE-/- mice by reducing whole blood viscosity (WBV) at high rate and improving erythrocyte function. In conclusion, LTC could ameliorate erythrocyte deformability and osmotic fragility through the reduction of lipid peroxidation on plasma and erythrocyte membranes in HFD-induced ApoE-/- mice, which supported the traditional uses of Chinese dragon's blood as an effective agent for improving blood microcirculation in hypercholesterolemia.

Figure 1

High-performance liquid chromatogram of LTC at 280 nm. 1: 7,4′-dihydroxyflavone; 2: loureirin D; 3: 7,4′-dihydroxyhomoisoflavanone; 4: loureirin C; 5: 3,4′-dihydroxy-5-methoxystilbene; 6: 5,7-dihydroxy-4′-methoxy-8-methylflavan; 7: 4-hydroxy-2,4′-dimethoxydihydrochalcone; 8: loureirin A; 9: loureirin B; and 10: pterostilbene.

Figure 2

Chemical structures of the main components in LTC.

Figure 3

TC, TG, and LDL-c levels and MDA levels in plasma of ApoE−/− mice obtained after six weeks of drugs treatment. (a) TC, TG, and LDL-c levels in plasma. (b) MDA levels in plasma. The results reflect the outcome of experiments conducted on 9~10 mice for each measurement. ^∗ P < 0.05 versus HG; ^∗∗ P < 0.01 versus HG; ^∗∗∗ P < 0.001 versus HG. EG: ezetimibe-treated group, HFD: high fat diet, HG: HFD-induced group, LDL-c: low density lipoprotein cholesterol, LTC: Longxuetongluo Capsule, LTC100 group: low dose LTC-treated group, LTC200 group: middle dose LTC-treated group, LTC300 group: high dose LTC-treated group, MDA: Malondialdehyde, TC: total cholesterol, and TG: triglycerides.

Figure 4

Deformability of erythrocytes in ApoE−/− mice. (a) The deformation curve integral area of erythrocytes. (b) The (DI)_max of erythrocytes. The results reflect the outcome of experiments conducted on 9~10 mice for each measurement. ^∗ P < 0.05 versus HG; ^∗∗ P < 0.01 versus HG. DI: erythrocyte deformation index, HFD: high fat diet, HG: HFD-induced group, LTC: Longxuetongluo Capsule, LTC100 group: low dose LTC-treated group, LTC200 group: middle dose LTC-treated group, and LTC300 group: high dose LTC-treated group.

Figure 5

Osmotic fragility of erythrocytes in ApoE−/− mice of HG, LTC groups (100, 200, and 300 mg/kg), and EG: the percentages of hemolysis (%) rate against osmotic pressures (mOsm/kg). The results reflect the outcome of experiments conducted on 9 ~ 10 mice for each measurement. ^∗∗∗ P < 0.001 versus HG; ^# P < 0.05 versus EG. HFD: high fat diet, HG: HFD-induced group, LTC: Longxuetongluo Capsule, LTC100 group: low dose LTC-treated group, LTC200 group: middle dose LTC-treated group, and LTC300 group: high dose LTC-treated group.

Figure 6

Morphological changes of erythrocytes in HFD-induced and LTC-treated mice observed with scanning electron microscope. (a) Erythrocytes of HFD-induced mice deformed with protrusions or irregular appearances. (b) Erythrocytes of LTC-treated (300 mg/kg) mice presented the more typical discocytes with rare deformed cells. (c) Erythrocytes of ezetimibe-treated mice presented the more typical discocytes with rare deformed cells.

Figure 7

Effects of LTC on the association of erythrocyte skeleton. (a) The erythrocyte membrane ghosts were separated by SDS-PAGE. Here was the identification of the major erythrocyte membrane proteins. ((b), (c), and (d)) The quantitative results of the ratio of protein 4.1, band 3, and spectrin to actin. The results reflect the outcome of experiments conducted on 6 ~ 10 mice for each measurement. HFD: high fat diet, HG: HFD-induced group, LTC: Longxuetongluo Capsule, LTC100 group: low dose LTC-treated group, LTC200 group: middle dose LTC-treated group, and LTC300 group: high dose LTC-treated group.

Figure 8

Effects of LTC on the lipid content and lipid peroxidation of erythrocyte membrane. (a) FC of erythrocyte membranes among HG, LTC groups (100, 200, and 300 mg/kg), and EG. (b) Effects of LTC on lipid peroxidation in erythrocyte membranes. ^∗ P < 0.05 versus HG; ^∗∗∗ P < 0.001 versus HG. FC: free cholesterol, HFD: high fat diet, HG: HFD-induced group, LTC: Longxuetongluo Capsule, LTC100 group: low dose LTC-treated group, LTC200 group: middle dose LTC-treated group, LTC300 group: high dose LTC-treated group, MDA: Malondialdehyde.