Therapeutic effects of Lingguizhugan decoction in a rat model of high-fat diet-induced insulin resistance

BACKGROUND Lingguizhugan(LGZG)decoction is a widely used classic Chinese medicine formula that was recently shown to improve high-fat diet(HFD)-induced insulin resistance(IR)in animal studies.AIM To assess the therapeutic effect of LGZG decoction on HFD-induced IR and explore the potential underlying mechanism.METHODS To establish an IR rat model,a 12-wk HFD was administered,followed by a 4-wk treatment with LGZG.The determination of IR status was achieved through the use of biochemical tests and oral glucose tolerance tests.Using a targeted metabolomics platform to analyze changes in serum metabolites,quantitative real-time PCR(qRT-PCR)was used to assess the gene expression of the ribosomal protein S6 kinase beta 1(S6K1).RESULTS In IR rats,LGZG decreased body weight and indices of hepatic steatosis.It effectively controlled blood glucose and food intake while protecting islet cells.Metabolite analysis revealed significant differences between the HFD and HFDLGZG groups.LGZG intervention reduced branched-chain amino acid levels.Levels of IR-related metabolites such as tryptophan,alanine,taurine,and asparagine decreased significantly.IR may be linked to amino acids due to the contemporaneous increase in S6K1 expression,as shown by qRT-PCR.CONCLUSIONS Our study strongly suggests that LGZG decoction reduces HFD-induced IR.LGZG may activate S6K1 via metabolic pathways.These findings lay the groundwork for the potential of LGZG as an IR treatment.

Protective effects of Lingguizhugan decoction on amyloid-beta peptide (25-35)-induced cell injury Anti-inflammatory effects

In the present study, a human neuroblastoma cell line （SH-SY5Y） and BV-2 microglia were treated with amyloid-β peptide （25-35）, as a model of Alzheimer＇s disease, to evaluate the protective effects of 10-3-10-8 g/mL Lingguizhugan decoction and to examine the underlying anti-inflammatory mechanism. Lingguizhugan decoction significantly enhanced the viability of SH-SY5Y cells with amyloid-β peptide-induced injury, and lowered levels of interleukin-1β, interleukin-6, tumor necrosis factor-a and nitric oxide in the culture supernatant of activated BV-2 microglia. The effects of 103 g/mL Lingguizhugan decoction were more significant. These results suggest that Lingguizhugan decoction can protect SH-SY5Y cells against amyloid-β peptide （25-35）-induced injury in a dose-dependent manner by inhibiting overexpression of inflammatory factors by activated microglia.

Effect of alternate-day-fasting combined with Lingguizhugan Decoction on blood lipid profiles of hyperlipidemic rats

Objective： To explore the effects of alternate-day-fasting （ADF） therapy combined with Lingguizhugan Decoction （LD）on blood lipid profiles of hyperlipidemic rats. Methods： Rats were randomly assigned into high-fat-diet （HF） group andnormal-diet （ND） group. Hyperlipidemic rats fed with high-fat-diet for 5 weeks were randomly divided into ADF group,alternate-day-fasting with LD （ALG） group and model control （MC） group. The rats in ALG and ADF group weredeprived of food for 24 h every other day for 4 weeks. Rats in ALG group were administrated with LD at fasting day.After 4 weeks of ADF therapy, plasma TC, TG, LDL-c and HDL-c were measured in each group. Expression of miR-143and PPAR-γ protein from adipose was also analyzed. Results： When compared with MC group, after 4 weeks of ADF orcombined ADF and LD therapy, the body weight was evidently reduced in ADF and ALG groups （P = 0.028, P = 0.036by wk 8）. The levels of plasma TC and TG decreased in ADF group and ALG group, which were significantly lower thanthose in MC group （P 〈 0.001, P = 0.045; P 〈 0.001, P = 0.005）. However, the body weight and level of TC and TG inALG group showed non-statistical difference in comparison with ADF group （ALG vs. ADF, P 〉 0.05）. Expression ofmiR-143 and PPAR-γ were higher in MC group than that in NC group （P 〈 0.001）. Compared with MC group,expression of miR-143 and PPAR-γ were significantly decreased in ADF （P = 0.038, P = 0.015） and ALG （P = 0.007, P〈 0.001） groups. When compared with ADF group, expression of miR-143 and PPAR-γ were significantly decreased inALG （P = 0.041, P = 0.046） group. Conclusion： ADF therapy alone not only reduced blood lipids, but also inhibitedmiR-143 and PPAR-γ protein expression in visceral adipose tissue. However, LD couldn’t reduce the levels of bloodlipid profiles more effectively than using ADF alone. Perhaps the effects of LD combined with ADF in the prevention ofhyperlipidemia need further exploration.

Effects of Lingguizhugan Decoction on α-SMA and collagen synthesis in rat myocardial fibroblasts induced by transforming growth factor-β_(1)

Objective:To explore the protective effect of Linggui Zhugan Decoction(LGZGD)medicated serum on myocardial fibrosis induced by transforming growth factor-β1(TGF-β1).Methods:Using enzyme digestion method,combined with differential adherence to isolate and culture Sprague-Dawley(SD)suckling mouse Cardiac fibroblasys(CFB)in vitro.Divided into:blank group,blank rat serum group,model group,and LGZGD medicated serum group(5%、10%、20%).Except for blank group and blank rat serum group,they were stimulated with 5 ng/ml TGF-β1 for 12 hours,and then then intervene with LGZGD medicated serum(5%、10%、20%)and continue to culture for 24 hours.Use immunofluorescence and Western blot(WB)to detect the expression ofα-smooth muscle actin(α-SMA),Enzyme-linked immunosorbent assay(ELISA)and WB to detect type Ⅰ collagen(Collagen Ⅰ),type Ⅰ collagen(Collagen Ⅲ)and fibronectin(FN)expression.Results:Compared with the blank group,the expressions of Collagen Ⅰ,Collagen Ⅲ,α-SMA and FN in the model group were significantly increased(P<0.01);Compared with the model group,the expressions of Collagen Ⅰ and Collagen Ⅲ in each concentration group of the experiment were significantly reduced(P<0.01);the expression ofα-SMA and FN were significantly reduced(P<0.01).Conclusions:LGZGD has an inhibitory effect on collagen synthesis and the expression ofα-SMA and FN,indicating that the anti-fibrosis effect of LGZGD is related to it.

Effects of modified Lingguizhugan decoction combined with weekend fasting on metabolic syndrome

OBJECTIVE:To assess the therapeutic effect of weekend fasting and administration of a modified Lingguizhugan decoction on metabolic syndrome(MetS).METHODS:Twenty-one patients with MetS were recruited from the First Affiliated Hospital of Sun Yat-Sen University.Fasting plasma glucose(FPG),30-min and 2-h post-prandial blood glucose(PG),fasting serum insulin(FINS),blood pressure(BP),body mass index(BMI),waist circumference(WC),homeostasis model assessment for insulin resistance index(HOMA-IR),and levels of triglyceride(TG),total cholesterol(TC),low-density lipoprotein-cholesterol(LDL-C),and high-density lipoprotein cholesterol(HDL-C)were tested.Patients were allowed to drink only water and a Chinese herbal decoction during weekends.All samples were tested again after 12 weeks of treatment.RESULTS:FPG,30-min PG,2-h PG,FINS,LDL-C,systolic BP,diastolic BP,BMI,WC,and HOMA-IR de-creased significantly(P<0.05)compared with before treatment.Levels of TG,TC,and HDL-C did not change significantly.CONCLUSION:Weekend fasting improved glucose metabolism,lowered BP,reduced LDL-C levels,BMI,and WC.These data suggest that weekend fasting may be an effective therapy for MetS by protection against coronary atherosclerosis.

Effect of a combination of calorie-restriction therapy and Lingguizhugan decoction on levels of fasting blood lipid and inflammatory cytokines in a high-fat diet induced hyperlipidemia rat model

OBJECTIVE:To combine calorie-restriction therapy(CRT) with Traditional Chinese Medicine(TCM) using Lingguizhugan Decoction and note the effects on expression of inflammatory cytokines[tumor necrosis factor(TNF)-α,high-sensitivity C-reactive protein(hs-CRP),interleukin(IL)-6]in high-fat-diet induced hyperlipidemia in rats.METHODS:Forty rats were divided randomly and equally into four groups:control group A(regular feeding for 5 weeks);model group B(high-fat diet for 5 weeks),calorie-limited group C(high-fat diet for 5 weeks) and TCM calorie-limit group D(high-fat diet for 5 weeks).After modeling,groups C and D were fasted for 3 days:group C with simple fasting,and group D with TCM fasting.The motion as well as changes in color,body weight,food intake,plasma lipids[low-density lipoprotein-cholesterol(LDL-C),high-density lipoprotein-cholesterol,total cholesterol(TC) and triglyceride(TG) along with TNF-α,hs-CRP and IL-6]were measured before and after intervention.RESULTS:Modeled rats were established after five weeks.After 3 days of fasting,compared with group B,blood lipid levels(TG,TC,LDL-C) of groups C and D decreased dramatically.Those of group D decreased more obviously than those of group C.However,the values of TNF-α,hs-CRP and IL-6 showed no obvious difference between each of the three treatment groups.CONCLUSION:CRT can reduce fasting blood lipid levels in rats,but not by reducing the levels of inflammatory cytokines.