Effect Of Spleen Deficiency On Liver Lipid Deposition In Rats With Hyperlipidemia And Its Mechanism

Purpose:Traditional Chinese medicine thought, spleen governing transportation and transformation, dysfunction of the spleen in transportation can cause abnormal lipid metabolism. This study take the Chinese medicine pathogenesis as the breakthrough point, copy spleen deficient hyperlipidemia rat model and discusse the effect and mechanism of spleen deficiency on blood lipid disorders and liver lipid deposition in hyperlipidemia rats of pi deficiency under the guide of traditional Chinese medicine theory. Methods: 60 SPF SD male rats, weight(300±10)g, started to be experimented after adapting to the environment for 7 days. According to random number table method, experiments with rats were assigned into blank control group, hyperlipidemia group and spleen deficiency combined hyperlipidemia group. Rats in blank control group were filled with ordinary pellet feed; Rats in hyperlipidemia group were filled with high fat feed; Rats in spleen deficiency combined hyperlipidemia group were treated with swimming excessive labour combined high fat feed. The modeling lasted for 60 days. Using phloroglucinol method to test D-xylose excretion rate; Levels of serum cholesterol(TC), triglyceride(TG), low density lipoprotein cholesterol(LDL-C), and high density lipoprotein cholesterol(HDL-C) were detected by fully automatic biochemical analyser; Using HE stain to observe the liver morphology change; Using oil red O stain to observe the liver lipid deposition. Levels of liver TC and TG were detected by COD-CE-PAP and GPO-PAP method respectively; Using real-time RT-PCR and Western blotting technolog to detect the mRNA and protein expression level of HMGCR, HL, CYP7A1, LDL-R. Statistical processing: Analyzed by SPSS 17.0 software package with the One-Way ANOVA method; Statistical results expressed in( x ±S)and with P < 0.05 for the difference was statistically significant. Results: 1. The influence of pi deficiency on blood lipid, AMY activity and D-xylose excretion rate of hyperlipidemia rats.The results show: Comparing with blank control group, TC, TG levels increased significantly(P<0.01), HDL-C level and AMY activity decreased significantly in hyperlipidemia group and spleen deficiency combined hyperlipidemia group(P < 0.05, P<0.01); Comparing with hyperlipidemia group, TC and LDL-C increased significantly(P<0.05, P<0.01), HDL-C, AMY and D-xylose excretion rate decreased significantly in spleen deficiency combined hyperlipidemia group(P<0.05, P<0.01). 2. The influence of pi deficiency on liver pathological morphology and lipid deposition of hyperlipemia rats.The results show: Comparing with blank control group, A large amount of fatty degeneration and lipid deposition occurred in liver of hyperlipidemia group and spleen deficiency combined hyperlipidemia group. 3. The influence of pi deficiency on liver lipid of hyperlipidemia rats.The results show: Comparing with blank control group, TC and TG levels increased significantly in hyperlipidemia group and spleen deficiency combined hyperlipidemia group(P<0.05, P<0.01); Comparing with hyperlipidemia group, TC and TG levels increased significantly in spleen deficiency combined hyperlipidemia group(P<0.01). 4. The influence of pi deficiency on HMGCR, HL, CYP7A1 and LDL-R mRNA expression in hyperlipidemia rats liver.The results show: Comparing with blank control group, HMGCR, HL, CYP7A1 and LDL-R mRNA expression level decreased significantly in hyperlipidemia group and spleen deficiency combined hyperlipidemia group(P<0.05, P<0.01); Comparing with hyperlipidemia group, HL, CYP7A1 and LDL-R mRNA expression level decreased significantly in spleen deficiency combined hyperlipidemia group(P<0.05?P < 0.01), HMGCR mRNA expression had no significant change. 5. The influence of pi deficiency on HMGCR, HL, CYP7A1 and LDL-R protein expression in hyperlipidemia rats liver.The results show: Comparing with blank control group, HMGCR, HL, CYP7A1 and LDL-R protein expression level decreased significantly in hyperlipidemia group and spleen deficiency combined hyperlipidemia group(P<0.05, P<0.01); Comparing with hyperlipidemia group, HL, CYP7A1 and LDL-R protein expression level decreased significantly in spleen deficiency combined hyperlipidemia group(P<0.05, P<0.01), HMGCR protein expression had no significant change. Conclusions: 1. Spleen deficient hyperlipidemia rat model have been builted successfully by the method of swimming excessive labour combined high fat feed. 2. Spleen deficiency aggravate blood lipid disorders and liver lipid deposition of hyperlipidemia rats. 3. Spleen deficiency affect mRNA and protein expression about cholesterol intake and transformation in liver.