Clinical Observation And Experimental Study On Treating Fatty Liver By The Means Of Strengthening Spleen, Resolving Phlegm And Promoting Blood Circulation

The paper, based on the clinic experience of tutor and professor Zhang huichen,think the pathobiology and action mechanism Fatty Liver is mainly blood stasis,qi-stagnation and damp stagnation, which often become mutual causality or interaction.So, we apply the means of strengthening spleen, resolving phlegm and promoting bloodcirculation to treat the Fatty Liver, and make up prescription according to the idea. TheModified Bupleurum and Peony plus two Vintage Herbs Decoction, (composed byPoria,Pinellia Tuber,Tangerine Peel,Bupleurum root,white Peony root,AlismatisRhizome,siberian solomonseal rhizome,Moutan bark,safflower,red Sage root,Hawthorn Fruit,fleece-flower root, etc.) has the function of strengthening spleen,regulating liver and removing stasis and phlegm. And it has been proved significantlyeffective in treating fatty liver through years of clinical observation. For the purpose ofsummarizing its treating effect, we made random-comparison (single blind) clinicalstudy with strict design. In order to further probe its action mechanism, we duplicatedthe fatty liver animal model with fatty emulsion, at the same time, we interfered theanimal with medicine. And we studied the action mechanism of this formula inregulating lipid metabolism and protecting liver function through observing its effect onTC, TG, HDL-C, LDL-C, AST and ALT. We also studied its compatibility of medicinesby literature, clinical and experimental study.PartⅠ: Literature overviewFatty Liver is accumulation of fat in liver because of sorts of reason when 5% ofweight or 1/3 in area. In TCM, Fatty Liver belongs to the categories of "mass" or"hypochondria pain", etc.. In China, the incidence of fatty liver increased from 10% in 80s' to 15% in 90s'. Especially, during recent ten years, the incidence of fatty liverincreased markedly, which lead to high incidence of liver fibrosis up to the 25%. So, itis great social significance to further research the fatty liver and seek for effectivecurative method.The research of ancient and up-dated medical literature showed that hepaticstagnation, spleen impairment due to diet, damp stagnation due to spleeny asthenia,qi-stagnating and blood stasis is the main factor of fatty liver. In TCM, applying themeans of strengthening spleen, resolving phlegm and promoting blood circulation totreat the Fatty Liver usually acquired good effect.PartⅡ: Clinical observation on treating fatty liver by the means ofstrengthening spleen, resolving phlegm and promoting blood circulation1 MethodsIt was established strict diagnosis criteria, inclusion criteria, exclusion criteria andeffective evaluation criteria firstly. 74 qualified cases were divided into two groups atrandom. 38 cases in treatment group were treated with Modified Bupleurum and Peonyplus two Vintage Herbs Decoction and diet control. Other 35 cases in control groupwere treated with Xiangsha Liujunzi Detoct. The therapeutic effect was observed in twogroups after treatment single-blindedly. And we made observation on thepatients' pre-treatment and post-treatment changes in clinical symptoms, bloodfat, ultrasonic image of liver, liver function after one treatment course (3 month).2 Results2.1 changes of body weightThe pre-treatment and post-treatment body weight reduced in two groups, whichshowed two remedy can reduce weight. The body weight of treatment group wasmarkedly reduced than the control group, but no significantly lowers than the controlgroup.2.2 Comparison of two groups on fatty liver patients2.2.1 Changes of clinical symptomsThe effective rate of 38 cases of treat group on clinical symptoms was 92.32%,while that of 36 cases of control group was 61.22%, which showed that treat group ismuch superior to control group in improving the clinical symptoms.Among 38 cases in the treatment group, 18 cases got marked effect, 12 cases goteffect, 8 cases got no effect, and the whole effective rate was 47.35%; among 36 casesin the control group, 2 cases got marked effect, 17 cases got effect, 17 cases got noeffect and the whole effective rate was 5.56%. Therefore the treatment group was significantly superior to the control group (P＜0.01).2.2.2 Blood serum assay between two-group patientsThe pre-treatment average value of TG, TC of treatment group was(248.92±20.72mg/dl,257.66±39.33mg/dl)respectively, and changed into (198.87±12.74mg/dl,158.58±20.51mg/dl) after treatment; The average value of TG, TC of control groupbefore treatment was (259.06±24.50mg/dl,245.47±68.04mg/dl)respectively andchanged into (242.39±31.56mg/dl,224.50±43.90mg/dl). The post-treatment averagevalues of TG, TC in both groups were significantly reduced than pre-treatment values(P＜0.05 or P＜0.01); and the average value of treatment group was lower than controlgroup (P＜0.01).2.2.3 Changes of ultrasonic examination between two-group patientsThe pre-treatment and post-treatment ultrasonic examination showed the treatgroup patients superior to the control group2.2.4 Changes of liver function between two-group patientsThe post-treatment value (37.18±6.69U/L,42.39±8.85U/L) of GOT and GPT intreatment group was significantly reduced than pre-treatment average value(52.24±9.94U/L,66.37±18.29U/L) (P＜0.01); the post-treatment value(48.61±11.70U/L,61.39±13.16U/L) of GOT and GPT in control group was alsosignificantly reduced than pre-treatment average value (51.28±14.40U/L,64.31±14.59U/L) (P＜0.01). However, the post-treatment average values of GOT and GPTof treatment group was significantly lower than control group (P＜0.01 or P＜0.05)..PartⅢ: Experimental study on treating fatty liver by the means ofstrengthening spleen, resolving phlegm and promoting blood circulation1 MethodsIn experimental study, 60 healthy male SD rats were divided into 4 groups atrandom, namely, (1) normal control group (normal group for short): Saline was given;(2) Model control group (model group for short): Saline was given; (3) ModifiedBupleurum and Peony plus two Vintage Herbs Decoction group (4) Xiangsha LiujunziTang control group (control group for short). Rats in each group were givenstomach-perfusion once a day and every time the drug volume was calculated by1ml/100g. Except for the ordinary forage in normal group, the rest of groups were allfed with fatty emulsion.Meanwhile control drug or treatment drug was given in each treatment group andSaline was given to normal and model group, which lasted for 1 weeks of stomach-perfusion. Confirmed the formation of fatty liver, Then 12 hours after fastingof the last time, the rats were cut off from the head and the their blood were drawn out,the blood sample was made centrifugation with low temperature and the blood serumwas isolated. Finally the contents of TC, TG, HDL-C, and LDL-C were measured withenzyme method; the contents of AST and ALT were measure with Lai's method. Andthe animal's liver was sectioned and removed simultaneously, 1cm×1cm liver tissuewas removed at 0.5cm from the biggest lobe to the border, fixed with formaldehyde andembedded with common paraffin, sliced and made HE staining for morphologicalobservation with photo scope. In addition, part of liver tissue (1.0-1.5g) was made into10% liver tissue homogenate and made 10-minute centrifugation with the condition of4℃4000r/min. And then the supernatant fluid was drawn out for measurement ofcontents of TC, TG, HDL-C and LDL-C by 722-type raster spectrophotometer.2 Results2.1 Changes of serum lipidThe contents of TC, HDL-C,LDL-C in serum of model rats (4.30±0.78, 0.84±0.13, 1.34±0.34) were significantly increased than normal group (2.35±0.21, 0.67±0.09,0.39±0.13) (P＜0.01); The content of HDL-C in model group was abnormally reducedthan the normal group (P＜0.01), which showed that fatty liver rats had significantblood-fat metabolic disorder. After the intervention of medicine, the contents of TC, TG,HDL-C,LDL-C in serum of each treatment group were significantly reduced andHDL-C content was increased. There was significant difference between treatmentgroup and model group (P＜0.01). And the contents of TC, HDL-C,LDL-C in treatmentgroup (2.61±0.29, 0.78±0.07, 0.67±0.17) were obviously reduced than the model group(4.30±0.78, 0.84±0.13, 1.34±0.34) (P＜0.05 or P＜0.01).2.2 Atheromatous Index (AI)The post-treatment value (0.88±0.23) of AI in treatment group was significantlyreduced than pre-treatment average value (2.10±0.55) (P＜0.01); The post-treatmentvalue (1.66±0.40) of AI in control group have no changes than pre-treatment averagevalue (1.65±0.69) (P＜0.01). The post-treatment average values of AI of treatment groupwas significantly lower than control group (P＜0.01).2.3 Changes of liver functionThe contents of ALT and AST in the rats' serum of model group (215.6±47.3,281.6±114.1) was markedly increased than the normal group (50.5±4.3,105.2±7.4)(P＜0.01). After the intervention of medicine, the contents of ALT and AST in the rats' serum of each treatment group were all obviously reduced, and had significantdifference with model group (P＜0.01). The ALT and AST contents in the rats' serum oftreatment group (76.5±6.1,133.8±8.9)was obviously reduced than the control group.2.4 Changes of liver lipidThe contents of TC, TG in liver tissue of model rats (166.70±17.98,426.48±43.19) were obviously increased than normal group(46.71±9.18, 45.34±9.55)(P＜0.01), which showed that fatty liver rats had obvious hepatic lipid metabolicdisorder.After the intervention of medicine, treatment group and positive drug controlgroup could all significantly reduced the contents of TC, TG in liver tissue, and all hadsignificant difference with model group (P＜0.01). And the contents of TC, TG in livertissue of treatment group (63.62±5.70, 88.90±10.29) were obviously reduced than thecontrol group (P＜0.01).2.5 Effect on liver tissue morphology of fatty liver ratAs for the rats in model group, they have unclear limit of hepatic lobules anddisordered arrangement of hepatic cord; major part of the hepatic sinus disappeared;majority of the hepatic cells appeared severe fatty degeneration, especially in the areaaround the lobules; hepatic cells became extremely swollen and formed round shape;lipid droplet vacuoles (fatty degeneration) in different sizes and numbers was visibleinside the cytoplasm, and sometimes the lipid droplets merged together and squeezedthe nucleus to the cell side, which made it look very similar with fatty cells; the rest ofthe hepatic cells had focal necrosis and hepatic interstitials had no fibrosis changes;While as for the rats in treatment group, they had very clear structure of hepatic lobulesand sinuses; the cells arranged orderly; lipid droplet vacuoles basically disappeared; themorphology and structure of nucleus became normal and regenerated hepatic cells werevisible; and as for the rats in the control group, they had clear structure of hepaticlobules; hepatic sinus became mildly expanded; fatty degeneration of hepatic cellsbecame mild; scanty lipid droplet vacuoles were still visible; the structure of nucleuswas comparatively clear; Compared with the model group, the degree of fattydegeneration in each drug group got significant improvement.The Modified Bupleurum and Peony plus two Vintage Herbs Decoction hadmarked function in regulating blood fat and protecting liver, it manifested like this: itcould inhibit the form of fatty liver; inhibit the absorption of exogenous lipid; inhibitthe synthesis of triglyceride and cholesterol; affect the distribution, transportation andremoval of lipid; accelerate the discharge of cholesterol and remove the deposition of fatty liver; improve the hepatic lipid metabolism.The Modified Bupleurum and Peony plus two Vintage Herbs Decoction couldobviously improve the liver tissue lesion degree of hyperglycemia-fatty liver, itmanifested like this: after the Chinese medicine was given, the hepatic lobule and sinusgot clear structure; the cells arranged orderly, no obvious lipid droplet vacuoles insidethe hepatic cells; only scanty hepatic ceils appeared mild swollen, and that proved thecapsule had apparent anti-liver injury function.PartⅣConclusion1 Clinical study indicated:Disharmony between liver and spleen and mixture of phlegm and stasis are themajor pathogenesis of fatty liver, and therefore the effective treatment method shouldbe to strengthen spleen, regulate liver and remove both phlegm and stasis. The clinicalstudy with strictly-designed random comparison (single blind) indicated that TheModified Bupleurum and Peony plus two Vintage Herbs Decoction could significantlyimprove the patients' clinical symptoms, ultrasonic image, blood fat (TG, TC, HDL-C,and LDL-C) and serum AST and ALT. Compared the therapeutic effect, total effectiverate and marked effective rate in treatment group were 78.95% and 47.35% respectively.Those in control group were 52.78% and 5.56% respectively. The therapeutic effect ofthe former was better than that the latter, with significantly difference between twogroups. It is concluded that Modified Bupleurum and Peony Plus Two Vintage HerbsDecoction can decrease the levels of serum GPT, TC and TG and improve fattydegeneration in the liver observed by ultrasound, its effect is obviously better than thatin control group treated with diet control. and that fully proved that The ModifiedBupleurum and Peony plus two Vintage Herbs Decoction could be the highly effectivedrug for fatty liver.2 Experiment indicated:2.1 The Modified Bupleurum and Peony plus two Vintage Herbs Decoction hadmarked function in regulating blood fat and protecting liver, it manifested like this: itcould inhibit the form of fatty liver; inhibit the absorption of exogenous lipid; inhibitthe synthesis of triglyceride and cholesterol; affect the distribution, transportation andremoval of lipid; accelerate the discharge of cholesterol and remove the deposition offatty liver; improve the hepatic lipid metabolism.2.2 The Modified Bupleurum and Peony plus two Vintage Herbs Decoction couldobviously improve the liver tissue lesion degree of fatty liver, it manifested like this:after the capsule was given, the hepatic lobule and sinus got clear structure; the cells arranged orderly, no obvious lipid droplet vacuoles inside the hepatic cells; only scantyhepatic cells appeared mild swollen, and that proved the capsule had apparent anti-liverinjury function.During the experiment, we observed the model rats with different components andprobed into the action mechanism of the Recipe on fatty liver by observing the effect ofthe Recipe on rat fatty liver from inhibiting the absorption of exogenous lipid,inhibiting the synthesis of endogenetic cholesterol, affecting the distribution,transportation and removal of lipid and accelerating the discharge of cholesterol andremoving the deposition of fatty liver, etc.. The recipe can markedly reducetransaminase of liver rise level, while it has no marked effect on absorption of lipidfrom intestine. Because of the complexity of lipid metabolism, the recipe can acceleratethe discharge of cholesterol, remove the deposition of fatty liver and improve thehepatic lipid metabolism. The effect of the recipe is synergetic effect, not adding eachother simply, which prove the benefit of compatibility of medicines.In short, the experimental study confirmed the action mechanism of The ModifiedBupleurum and Peony plus two Vintage Herbs Decoction treating fatty liver byregulating the lipid metabolism, resisting the injury of lipid peroxidation, improving themicrocirculation of liver, and protecting injury of liver, which indicated the treatmentalsuperiority of whole adjustment with the traditional Chinese medicine.