Characterization Of TCM Pathological Features Of Metabolic Syndrome (MS) And Study Of The Effects Of Modified Zexie Recipe On MS And Its Mechanisms

Metabolic syndrome（MS） is an aggregation of a series of metabolic disorders,including hypertension, hyperglycemia, increased cholesterol and glycerol, andinsulin resistance, which increase the risk of developing cardiovascular disease anddiabetes. Traditional Chinese Medicine (TCM) defined that “Fat people have muchmore phlegm”, and “blood stasis always happens with phlegm concomitantly”，thusrecognizing phlegm-blood stasis syndrome as an important risk factor for MS.Modified Zexie Recipe is an empirical formula used for treating phlegm-blood stasissyndrome, and has been shown effective in strengthening spleen and dissipatingdampness. Our previous study showed that Modified Zexie Recipe decreased bloodglucose and glycerol levels, thus ameliorating the symptoms of obesity and diabetes,while the mechanisms of which remain unclear. In addition, the clinical effectivenessof Modified Zexie Recipe on MS still needs further investigation. Herby the presentstudy was undertaken to characterize the pathological features of MS using TCMtheory, and to investigate the effects of Modified Zexie Recipe in treating MS patientsand in treating an animal model of MS induced by high-fat-diet.Part I: Characterization of the TCM pathologic features of metabolic syndromeObjective:To characterize the TCM pathologic features of MS, and to identify themiscellaneous and progressive patterns of MS.Methods：Data from491MS patients was analyzed to identify the main niduses, featuresand nature of MS, with syndrome element differentiation method, and the patterns ofmiscellaneous and progressive tendency of MS. Results:（1）The niduses of MS are located in spleen, kidney, liver, heart and lung,mainly in the spleen (60.4%), kidney (58.2%) and liver (51.5%);（2）The sthenia-syndrome consists of phlegm, damp, blood stasis, hot and qistagnation,with phlegm, damp and blood stasis at a higher proportion of77.1%,65.9%and61.3%respectively;（3）The asthenia-syndrome consists of Yin deficiency, Qi deficiency, Yangdeficiency, blood deficiency and jing deficiency, with Yin deficiency and Qideficiency at a higher proportion of64.7%and60.0%respectively;（4）The mixed features of the syndrome: when presented in a descending orderfrom high to low, the distribution of frequency number of MS were intermingledTurbid Phlegm and Blood Stasis syndrome, dual Qi-yin deficiency syndrome, dual QiDeficiency of spleen and kidney, dual Qi-Damp stagnation syndrome accordingly.The mixing degree of the niduses and the nature in each syndrome type was different,as for the mixed niduses, Spleen was the most common nidus in the Spleen Qi andKidney Qi Deficiency syndrome, and the second nidus is Phlegm and Blood stasissyndrome and Qi-yin deficiency syndrome; Phlegm was the most in Turbid Phlegmand Blood Stasis syndrome, then in Qi and Damp Stagnation syndrome and Spleen Qiand Kidney Qi Deficiency syndrome; while stasis was the most in Turbid Phlegm andBlood Stasis syndrome, then in Qi-yin deficiency syndrome; Damp was the most inQi and Damp Stagnation syndrome, then in Turbid Phlegm and Blood Stasissyndrome and Qi-yin deficiency syndrome;（5）The evolution tendency of syndrome：The incidence at kidney and liverniduses increased with age; the occurrence of blood stasis, Yin deficiency, Qideficiency, Yang deficiency, and blood deficiency were also tended to increase withthe age；The incidence at spleen, kidney, and liver niduses was higher in female thanin male，so does the occurrence of blood stasis and blood deficiency;（6）Phlegm syndrome was positively correlated with the TG (r=0.149，P＜0.05),BMI (r=0.341, P＜0.01), abdominal circumference (r=0.309, P＜0.01)，while was negatively correlated with HDL (r=-0.159, P＜0.05); Damp syndrome had a positivecorrelation with the abdominal circumference（r=0.146，P＜0.05），and had anegative correlation with HDL（r=-0.146, P＜0.05）；There is no correlationshipbetween blood stasis with this metabolic factors；Conclusions:MS niduses are mainly located at spleen, kidney and liver. The feature of MS isintermingled deficiency and excess, with phlegm, damp and blood stasis presentedthroughout the course of the disease. Age and gender are two important factorsaffecting the syndrome elements of MS and the disease progression. The youthsuffered more from spleen deficiency, phlegm and blood stasis, while the elderlypeople suffered more from both of the spleen and kidney deficiency. Women suffermore from blood deficiency of liver and blood stasis compared to men.PartⅡ: Clinical investigation of Modified Zexie Recipe on MSObjective:To investigate the effects of Modified Zexie Recipe on integral of syndromeelements and biochemical index，and to evaluate its therapeutic effect.Methods：Seventy MS patients, diagnosed as phlegm-blood stasis syndrome with TCM,were recruited and were randomly divided into two groups: Modified Zexie Recipeintervention group and the control group. After8weeks treatment with ModifiedZexie Recipe or placebo, the syndrome element changes of phlegm and blood stasiswere evaluated by syndrome element integral, and the serum glucose and lipidmetabolic status were assessed by biochemical indicators.Results:（1）With Modified Zexie recipe treatment, the syndrome elements integral ofspleen, phlegm, damp, and stasis were significantly reduced, the intermingledsyndrome in the liver and Qi deficiency was also significantly decreased. The phlegmsyndrome was decreased in the control group, while to a less extend compared to thedecrease in Modified Zexie recipe group; （2）Modified Zexie Recipe treatment had no effect on BMI（P＞0.05）, whilesignificantly decreased total and visceral fat, decreased serum cholesterol and TGlevels, and ameliorated glucose tolerance and insulin resistance (P＜0.05).Conclusions:Modified Zexie recipe is effective in reducing integral of spleen, phlegm, dampand blood stasis, which work together to improve glucose-lipid metabolism andinsulin resistance, thus have therapeutic effects on MS.Part Ⅲ Study of the effects of Modified Zexie recipe on glucose and lipidmetabolism and its mechanisms in MS ratsObjective:To investigate the effects of Modified Zexie Recipe on glucose and lipidmetabolism in a rat model of MS induced by high-fat-diet and its mechanisms.Methods：Rats were fed on a normal chew or high-fat diet for8weeks, then were randomlydivided into four groups: Control group (fed on normal chew), HFD group, HFD withModified Zexie Recipe treatment at low and high dose groups. Eight weeks followingModified Zexie Recipe treatment, body weight, visceral fat, liver weight, bloodglucose and fats level were measured, glucose metabolism and insulin sensitivity wasevaluated by oral glucose tolerance test (OGTT) and intraperitoneal injection insulintolerance test (IPITT) respectively. Hepatic steatosis was measured byimmune-histochemistry method, Lipid metabolic genes were analyzed by Real-timePCR, while AMP-activated protein kinase (AMPK) expression was examined byWestern blot.Results:（1）Rats fed on high-fat diet developed abdominal obesity, insulin resistanceand Non-alcoholic fatty liver disease (NAFLD).（2）Modified Zexie Recipe treatment had no effect on body weight（P＞0.05）,but reduced visceral fat (peri-epididymal and peri-renal) accumulation, decreasedCHO and TG levers in blood, improved OGTT and IPITT (P＜0.05), and amelioratedheptosteatosis. （3）With Modified Zexie Recipe treatment, the expressions of the key lipolyticgenes including peroxisome proliferators-activated receptor γ co-activator1α(PGC-1α), Peroxisome proliferator-activated receptor γ (PPAR-γ) and α (PPAR-α),were up-regulated (P＜0.05), while the expressions of lipogenic genes includingsterol regulatory element-binding protein1c (SREBP-1c), fatty acid synthase (Fasn)and Liver fatty acid-binding protein (L-FABP) were down-regulated (P＜0.05). Inaddition, Modified Zexie Recipe activated AMP-activated protein kinase (AMPK) andpromoted Acetyl-CoA carboxylase (ACC) phosphorylation in the liver.Conclusions:In a rat mode of MS induced by high-fat-diet, Modified Zexie Recipe promoteslipolysis while inhibits lipogenesis, improves glucose tolerance and insulin resistance,thus restores the disrupted glucose and lipid metabolism, might through activation ofAMPK signaling pathway.