| Objectives: To observe the effect of the Spleen-strengthening and Phlegm-resolving Prescription on content of TG,TC,HDL-C and LDL-C in blood serum of rats with Hyperlipidemia and the expression of AQP3 and AQP4 in colon tissue.At last,to discuss the treatment function of Spleen-strengthening and Phlegm-resolving Prescription on HLP and its possible mechanism.Methods: Sixty healthy male SD rats were randomly divided into6 group: Normal control group,Model control group,Simvastatin contro l group,Low-dose of spleen-strengthening and phlegm-resolving prescript ion,Middle-dose of spleen-strengthening and phlegm-resolving prescriptio n,High-dose of spleen-strengthening and phlegm-resolving prescription,Hi gh-fat forage was used to replicate the hyperlipidemia model.Rats in co ntrol group were fed with ordinary forage,and the others were fed with high-fat forage.At the same time,the normal group and the model gr oup were given the right amount of normal saline and the rest of the g roups were given the treatment drugs or the control drugs,the dosage was calculated by 1ml/100 g.After 5 weeks later,the rats fasting can no t help but water for 24 hours after the last administration.All rats were anesthetized and their blood drawn out from femoral artery,centrifuged,and then isolated the serum in order to measure the content of TG,T C,HDL-C and LDL-C in blood serum.Simultaneously,the liver were to ok out quickly and the maximum leaf liver was put into 4% poly Form aldehyde Solution for pathological observation.The colon were cut 3 pi eces,one was thrown into 4% poly Formaldehyde Solution for the obse rvation of immunohistochemical means,others were put into nitrogen canister for the observation of RT-PCR and Western Blot.Results:1 Changes of content of TG,TC,HDL-C and LDL-C in serum of rat in each group rats.Compared with normal group,the contents of TC,TG and LDL-C in serum of model group were significantly increased(P<0.01),the contents of HDL-C content was obviously decreased,the results showed that the HLP rat model was successful.After treatment,the contents of TG,TC and LDL-C in every treatment group were lower than that in the model group and the content of HDL-C in every treatment groups increased.Compared with the control group,the contents of TG、TC、HDL-C、LDL-C in high-dose group had no statistical significance(P>0.05),the contents of TG,TC,LDL-C in the low-dose group and middle-dose group were higher,the content of HDL-C was lower(P<0.01 or P<0.05).The comparison in 3 doses groups of spleen-strengthening and phlegm-resolving prescription,the content of TG、TC、LDL-C in high-dose group lower than those in middle-dose group and low-dose group(P<0.01 or P<0.05);Compared with low-dose group,the content of TC in middle-dose group was decreased,but the contents of TG 、 HDL-C 、 LDL-C had no statistical significance(P>0.05).2 Changes of liver tissue morphology in every group.Liver cells of rats in normal group were arranged orderly in the shape of a regular hexagon,distributed around the central veins.The structure of hepatic sinusoid was normal,and no fat droplets and inflammatory infiltration.In model group,the liver cells of rats arranged disorderly and the nucleus were extruded from the position.There were a large number of lipid droplets and the hepatic sinusoid was congestion and edema.The structure of Liver cell in control group was clear,fat droplets decreased obviously and the structure of hepatic sinusoid improved,but existed necrosis and inflammatory infiltration.Compared with model group,the structure of 3 doses groups of spleen-strengthening and phlegm-resolving prescription were cleaner,the lipid droplets also decreased in varying degrees and the morphology of hepaticsinusoids tended to be normal,especially in the high-dose group.3 Changes of AQP3 expression in colon tissue of ratsImmunohistochemical method: In normal group,the positive expression of AQP3 were relatively less and the staining were lighter.In model group,the positive expression of AQP3 at caryon,cytoplasm and intestinal gland epithelial cells were relatively more,the color were deeper and there were also a little positive expression at connective tissue.Semi quantitative analysis showed that the expression of AQP3 in colon of model group were significantly higher(P < 0.01).After treatment,the expression of AQP3 in each treatment group were lower than that in model group(P<0.01).Among them,the expression of AQP3 in high,middle and low-dose groups were obviously lower than that in the control group(P<0.01),the expression of AQP3 in high-dose of group were obviously lower than that in the low and middle-dose groups(P<0.01),the expression of AQP3 in middle-dose group were lower than the low-dose group(P < 0.05).RT-PCR method: The expression of AQP3 mRNA in colon of rats in model group were obviously higher than that in normal group(P<0.01).After treatment,the expression of AQP3 mRNA in each treatment group were lower than that in the model group.Among them,the expression of AQP3 mRNA in high-dose group and middle-dose group obviously lower than that in control group,but the low-dose group had no statistical significance compared with the control group(P>0.05),the expression of AQP3 mRNA in high-dose group obviously lower than that in middle-dose group and low-dose group(P <0.01),and the expression of AQP3 mRNA in middle-dose group obviously lower than that in low-dose group.Western Blot method:The results showed that the expression of AQP3 in colon tissue of model group obviously higher than that in normal group(P<0.01).After treatment,the expression of AQP3 in each treatment group lower than that in model group(P<0.01).Among them,the expression of AQP3 in high-dose group and middle-dose group obviously lower than in control group,but the low-dose group had no difference compared with the control group(P>0.05),the expression of AQP3 in high-dose group obviously lower than that in middle-dose group and low-dose group(P < 0.01),and the expression of AQP3 mRNA in middle-dose group obviously lower than that in low-dose group(P<0.01).4 Changes of AQP4 expression in colon tissue of rats.Immunohistochemical method:In normal group,the positive expression of AQP4 were relatively less and the staining were lighter.In the model group,the positive expression of AQP4 in caryon,cytoplasm and intestinal gland epithelial cells were relatively more,the color were deeper and there were also a little positive expression in connective tissue.Semi quantitative analysis showed that the expression of AQP4 in colon of model group were obviously higher(P<0.01).After treatment,the expression of AQP4 in each treatment group were lower than that in model group(P< 0.01).Among them,the expression of AQP4 in high-dose group,middle-dose group and low-dose groups were obviously lower than that in control group(P<0.01),the expression of AQP4 in high-dose of group were obviously lower than that in the low-dose group and middle-dose group(P<0.01),the expression of AQP4 in middle-dose group had no statistical significance compared with low-dose group(P>0.05).RT-PCR method: The expression of AQP4 mRNA in model group obviously higher than that in normal group(P<0.01),After the intervention,the expression of AQP4 mRNA in each treatment group lower than that in model group.Among them,the expression of AQP4 mRNA in high-dose group,middle-dose group obviously lower than that in control group(P<0.01),but there were no statistical significance between low-dose group and control group.The expression of AQP4 mRNA in high-dose group were lower than that in middle-dose group and low-dose group(P < 0.01),the expression of AQP4 mRNA in middle-dose group were lower than that in low-dose group(P<0.05).Western Blot method: The expression of AQP4 in model group were higher than that in normal group(P<0.01).After treatment,the expression ofAQP4 in each treatment group were lower than that in model group.Among them,the expression of AQP4 in high-dose group,middle-dose group and low dose group were lower than that in control group(P < 0.01 or P < 0.05),the expression in high-dose group were lower than that in middle-dose group and low-dose group(P<0.05 or P<0.01),the expression in middle-dose group was lower than that in low-dose group(P<0.05).Conclusions:1 The content of TG、TC、LDL-C in serum of rats with HLP could be decreased and the HDL-C could be increased through the treatment of spleen-strengthening and phlegm-resolving prescription,the degree of fatty degeneration of the liver cell could be improved,it points out that the spleen-strengthening and phlegm-resolving prescription had a satisfying effect on regulating the blood fat and improving disorder of blood fat.2 Spleen-strengthening and phlegm-resolving prescription achieve the purpose of treating HLP through restraining the expression of AQP3,AQP4,AQP3 mRNA,AQP4mRNA,improving water accumulation because of colon reabsorb water overly,reducing the formation of phlegm dampness and eliminating the cause of HLP. |
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