Study On The Anti-Obesity Effect And Mechanism Of Shenheling Fermented By Limosilactobacillus Fermentum

With the improvement of living standards,obese people are on the rise and have gradually become an important issue threatening public health.At present,there are four main problems in the research on the weight loss of traditional Chinese medicine with the homology of fermented medicine and food.First,the research on compound fermentation is insufficient,mainly focusing on the fermentation of single traditional Chinese medicine such as ginseng and lotus leaf.The way of action is relatively simple,and it is difficult to solve the multiple abnormal problems associated with obesity at the same time.The second is the lack of research on the simultaneous improvement of efficacy and flavor by fermentation,mainly focusing on the improvement of efficacy by fermentation,ignoring the impact of flavor on consumer acceptance and adherence to long-term use;third,the mechanism of action of fermented medicine and food homologous Chinese medicine to lose weight is not comprehensive enough;fourth,there are few reports on the weight loss research of fermented medicine and food homologous Chinese medicine in the population,mainly focusing on the weight loss research of animal models,and the weight loss effect in the population needs to be further verified.In view of the above problems,based on the theory of traditional Chinese medicine,this paper first combined the weight-loss compound with the homology of medicine and food,screened lactic acid bacteria that could improve the anti-obesity characteristics and flavor of the compound,and optimized its fermentation conditions.Then,the anti-obesity characteristics of fermented compound in rats(intervention to lose weight and prevention of obesity)were studied,and its possible mechanism was explored from the aspects of obesityrelated gene expression level and intestinal flora structure.Finally,its weight loss effect was preliminarily evaluated in overweight/obese people.The main research contents and results are as follows.(1)Compatibility of medicine and food homologous weight loss compounds and exploration of the effect of fermentation on flavor and function.First,through literature research,the theory of weight loss in traditional Chinese medicine,the mechanism of weight loss in traditional Chinese medicine compounds and the experience of prescription were summarized,and the Shenheling(SHL)compound with the effects of tonifying qi and strengthening spleen,resolving phlegm and removing dampness was combined.SHL water extract(SHLE)was obtained by soaking at room temperature and hightemperature extraction.The screened Limosilactobacillus fermentum grx08 fermentation significantly increased the lipase activity inhibition rate and antioxidant capacity of SHLE by 1.60 and 1.26 times,respectively(p<0.05).Hexanal and hexanol were the main substances of the grassy flavor of SHLE,which were not detected after fermentation.Fermentation significantly increased the sensory score of SHLE(p<0.05).The L fermentum grx08 prepared by liquid-to-material ratio of 10:1(v/m),inoculation amount of 4.10%(v/v),and fermentation at 37℃ for 49.50 h had higher lipase activity inhibition rate(93.28%)and viable cell number(1.84×109 CFU/mL),and had better stability during the 36 d storage period.UPLC-MS/MS analysis showed that fermentation significantly upregulated the content of 53 anti-obesity substances,such as ginsenoside Rh3,Rg3,and dehydronuciferine(p<0.05).(2)To explore the anti-obesity(intervention in weight loss and prevention of obesity)characteristics of L.fermentum grx08 fermented Shenheling in rats.Intervention Weight Loss Test,four groups of obese rat models were established by a high-fat diet.The nonintervention was used as the obesity model group(HFD),the L.fermentum grx08 fermented Shenheling group(SHLF08)and the nonfermented SHLE with L.fermentum grx08 bacteria group(SHLa08)were used as the weight loss test group,the Shenling Baizhu Powder group(SBZ)was used as the positive control group,and the normal diet group(ND)was used as the blank control group.The model was established for 4 weeks and then intervened for 4 weeks.The results showed that compared with the HFD group,the body weight of female and male rats treated with SHLF08 was significantly reduced by 16.97%and 10.48%,respectively(p<0.01),and the levels of triglyceride(TG),total cholesterol(TC)and low density lipoprotein cholesterol(LDL-C)in serum and liver were significantly reduced(p<0.05),and level of high-density lipoprotein-cholesterol(HDL-C)was significantly increased in female and male rats(p<0.05);the body weight and serum TG,TC,LDL-C and HDL-C levels of the SHLa08 group were between those of the HFD group and SHLF08 group.The results showed that L.fermentum grx08 fermentation improved the weight loss effect of Shenheling.To prevent obesity,a high-fat diet model was established by feeding rats a high-fat diet.The nonintervention group was used as the high-fat diet model group(HFDP),the L.fermentum grx08 fermented Shenheling prevention group(SHLF08P)was used as the experimental group,and the normal diet group(NDP)was used as the blank control group.SHLF08 was administered to prevent obesity while high-fat feeding.The results of 8 weeks showed that compared with the NDP group,the body weight of rats in the HFDP group was significantly increased by 20.04%(p<0.05),and the liver and white fat index were significantly increased(p<0.05),formed obese rats.However,SHLF08 prevention significantly reduced the body weight,liver and white fat index of high-fat diet rats(p<0.05),and there was no significant difference between the normal diet group(p>0.05),indicating that L.fermentum grx08 fermented Shenheling can better prevent high-fat diet-induced obesity.(3)To explore the anti-obesity mechanism of L.fermentum grx08.The antiobesity mechanism was studied by RT-PCR and intestinal flora.The results showed that compared with the obese model group(HFD),SHLF08 intervention significantly reduced the mRNA expression of Neuropeptide Y(NPY)and Agouti-related protein(AGRP)in the hypothalamus of rats(p<0.01)and the content of NPY in serum(p<0.05),and significantly increased the content of glucagon-like peptide-1(GLP-1)in serum(p<0.001).The food intake of female and male rats was significantly reduced by 19.58%and 9.74%,respectively,compared with the HFD group(p<0.01).This shows that SHLF08 can reduce the food intake of rats by regulating appetite.SHLF08 intervention significantly increased the mRNA expression of farnesol X receptor(FXR),AMP-activated protein kinase(AMPK)and carnitine palmitoyltransferase-1(CPT-1)in the rat liver(p<0.05).The mRNA expression of sterol regulatory element binding protein lc(SREBP-1C)and stearoyl-CoA desaturase-1(SCD-1)was significantly decreased(p<0.05),indicating that SHLF08 may inhibit fat synthesis through the FXR/SREBP-1C/SCD-1 pathway.It is possible that the AMPK/CPT-1 pathway increases fatty acid β oxidation,thereby reducing the accumulation of body fat.SHLF08 intervention significantly increased the expression of peroxisome proliferatoractivated receptor γ(PPARγ)and uncoupling protein 1(UCP-1)mRNA in brown fat(p<0.05)and increased the expression of UCP-1 protein(p<0.0001)and the brown fat index(p<0.01),indicating that SHLF08 increased the expression of heat-related genes and proteins.Compared with the ND group,the ratio of Firmicutes/Bacteroidetes(F/B)and the abundance of Treponema in the HFD group were significantly increased(p<0.05),while the abundance of Lactobacillus and Muribaculaceae was significantly decreased(p<0.05).Compared with the HFD group,the SHLF08 group significantly reduced the F/B ratio and the abundance of Spirochaetota at the phylum level(p<0.05)while significantly increasing the abundance of Bacteroidota and Actinobacteriota(p<0.05).At the genus level,the abundance of Bifidobacterium,Akkermansia,Lactobacillus and Muribaculaceae was significantly increased(p<0.05),while the abundance of Treponema and was significantly decreased(p<0.05),thereby improving the imbalance of intestinal flora induced by a high-fat diet.(4)To preliminarily explore the anti-obesity effect of L.fermentum grx08-fermented Shenheling in obese/overweight people.Through an 8-week randomized,placebo-controlled trial,the anti-obesity effect was explored in obese/overweight college students.The experimental group was treated with L.fermentum grx08-fermented Shenheling,and the placebo group was treated with the corresponding placebo.The results showed that compared with the placebo group,the body weights of the male and female groups in the experimental group were significantly reduced by 2.59 kg and 3.29 kg,respectively(p<0.05),and the body fat was significantly reduced by 2.13 kg and 1.85 kg,respectively(p<0.05).Shenheling fermented by L.fermentum grx08 significantly reduced serum TC,TG and LDL-C levels in the male group(p<0.05).The levels of LDL-C and uric acid in the female group were significantly decreased(p<0.05),as were the scores of qi deficiency,phlegm dampness and damp heat constitution(p<0.05).It shows that Lfermentum grx08 fermented Shenheling can reduce the fat mass and weight of overweight/obese subjects,improve blood lipid levels,and reduce the degree of qi deficiency,phlegm dampness and damp heat in female subjects.