Acupuncture improved hepatic steatosis in HFD-induced NAFLD rats by regulating intestinal microbiota.

Background: Intestinal dysbiosis has been increasingly implicated in the pathogenesis of non-alcoholic fatty liver disease (NAFLD). Acupuncture has been shown to have beneficial effects on NAFLD, but the mechanism is not yet clear. This study explores the potential beneficial effects of acupuncture on intestinal microbiota in NAFLD. Methods: An NAFLD model in Sprague Dawley rats was established using a high-fat diet (HFD) for 10 weeks. NAFLD rats were randomly divided into control, model, and acupuncture groups. Following acupuncture treatment over 6 weeks, automated biochemical analysis was used to measure serum lipid metabolism parameters, including levels of alanine transferase, aspartate transferase, alkaline phosphatase, total cholesterol, triglycerides, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol. The level of serum inflammatory factors interleukin (IL)-6, IL-10, and tumor necrosis factor-alpha (TNF-α) were measured by enzyme-linked immunosorbent assay. The characteristics of steatosis were evaluated using quantitative computed tomography, hematoxylin and eosin staining, and Oil Red O staining in the liver, while the intestinal microbiota was determined using 16S rRNA gene sequencing. Results: Acupuncture decreased the systemic inflammatory response, ameliorated dyslipidemia, and improved liver function indexes in NAFLD model rats. Tomography and staining indicated that acupuncture reduced steatosis and infiltration of inflammatory cells in the liver. 16S rRNA analysis showed that acupuncture reduced the Firmicutes to Bacteroidetes (F/B) ratio, increased the abundance of microbiota, including Bacteroidales_S24-7_group, Prevotellaceae, Bacteroidaceae, Blautia, norank_f_Bacteroidales_S24-7_group, Bacteroides, and Prevotella_9, and decreased the abundance of Ruminococcaceae_UCG-014. Correlation analysis suggested a close correlation between lipid metabolism, inflammation factors, hepatic steatosis, and the changed intestinal microbiota. Conclusion: Acupuncture can significantly improve lipid metabolism and the systemic inflammatory response in HFD-induced NAFLD rats, potentially by regulating intestinal microbiota composition.

[Impact of electroacupuncture on liver lipid metabolism and hepatic Sirt1 and PPARγ expression in abdominal obese rats].

OBJECTIVE: To observe the impact of electroacupuncture (EA) on liver lipid metabolism and expression of hepatic sirtuin 1(Sirt1) and peroxisome proliferator activated receptor γ(PPARγ) of abdominal obese rats induced by high-fat diet. METHODS: Eighteen male SD rats were divided into blank control, model and EA groups (n＝6 per group). The abdominal obesity model was established by feeding the rats with high-fat diet for 12 weeks. EA (2 Hz/15 Hz, 1.5 mA) was applied to bilateral "Daimai"(GB26) for 20 min every time, once every other day for 8 weeks. Rats of the model group were also restrained for 20 min. The body mass and abdominal circumference were measured every week, and the contents of serum cholesterol (TC), triglyceride (TG), alanine transaminase (ALT), aspartate aminotransferase (AST) were detected by using an automated biochemical analyzer. Histopathological changes of the liver tissues were observed under microscope after oil red "O" staining. The expression of hepatic Sirt1 and PPARγ mRNAs and proteins were detected using quantitative real time PCR and Western blot, separately. RESULTS: After modeling, the body weight and abdominal circumference, and serum TC, TG, ALT and AST contents, and expression of hepatic PPARγ mRNA and protein were significantly increased (P<0.001, P<0.01, P<0.05), and the expression levels of hepatic Sirt1 mRNA and protein obviously down-regulated (P<0.05, P<0.01) in the model group. Following EA intervention, the increased body weight and abdominal circumference, and serum TC, TG, ALT and AST contents, and hepatic PPARγ mRNA and protein expression were remarkably suppressed (P<0.05, P<0.01), and the decreased hepatic Sirt1 mRNA and protein were remarkably up-regulated (P<0.001，P<0.05). The lipid droplets in hepatocytes were reduced in the EA group relevant to the model group. CONCLUSION: EA intervention can significantly improve the liver lipid metabolism of abdominal obese rats, which is possibly related with its effect in up-regulating the expression of hepatic Sirt1 mRNA and protein, and in down-regulating the expression of hepatic PPARγ mRNA and protein.

Acupuncture Regulating Gut Microbiota in Abdominal Obese Rats Induced by High-Fat Diet.

OBJECTIVE: To investigate the effects of acupuncture on metabolic health and gut microbiota dysbiosis in diet-induced abdominal obese model. MATERIALS AND METHODS: Male Sprague-Dawley rats were randomly distributed into normal chow diet (NCD) group and high-fat diet (HFD) group. After 12 weeks of HFD feeding, an abdominal obese rat model was established. The abdominal obese rats were further assigned to acupuncture group (n=7) and nontreated HFD group (n=7). Acupuncture was applied to bilateral GB 26 of rats for 8 weeks. Subsequently, the body weight, waist circumference (WC), visceral fat mass, and liver weight were measured weekly in all rats. Metabolic parameters such as total cholesterol, triglyceride, alanine aminotransferase, aspartate transaminase, and blood glucose were measured by an automatic biochemical analyzer. The serum levels of insulin (INS) were determined using Rat INS ELISA Kit. Analysis of gut microbiota was carried out by 16S rRNA gene sequencing. RESULTS: Acupuncture decreased the body weight, WC, and visceral adipose tissues of HFD-induced abdominal obese rats. In addition, insulin sensitivity, glucose homeostasis, and lipid metabolism were improved by this treatment. Furthermore, electroacupuncture effectively modified the composition of gut microbiota, mainly via decreasing Firmicutes/Bacteroidetes ratio and increasing Prevotella_9 abundance. CONCLUSIONS: Electroacupuncture can ameliorate abdominal obesity and prevent metabolic disorders in HFD-induced abdominal obese rats, via the modulation of gut microbiota.

[Acupuncture improves hepatic lipid metabolism by suppressing oxidative stress in obese nonalcoholic fatty liver disease rats].

OBJECTIVE: To investigate the effect of acupuncture of "Daimai"(GB26) on abdominal fat accumulation, lipid metabolism and hepatic oxidative stress in abdominal obese non-alcoholic fatty liver disease (NAFLD) rats. METHODS: male SD rats were divided into 3 groups: normal diet (normal, n＝8), high fat diet control (model) and acupuncture (n＝8/group in the latter 2 groups). The abdominal obese NAFLD model was established by feeding the rats with high fat diet for 12 weeks. EA (2 Hz/15 Hz, 1.5 mA) was applied to bilateral GB26 for 20 min, once every other day for 8 weeks. Rats of the model group were also restrained for 20 min as those in the EA group. The body mass and abdominal circumference were measured weekly, the isolated adipose tissues around the bilateral kidney and epididymis and the fresh liver were weighed. The contents of serum cholesterol (TC), triglyceride (TG), alanine transaminase (ALT), glutamic oxaloacetic aminotransferase (AST) were detected by using an automatic biochemical analyzer. The abdominal visceral fat distribution was acquired by CT scanning. The oxidative stress indexes of the homogenated liver tissues, such as malondialdehyde (MDA) was assayed using sodium thiobarbital (TBA) method, and the activity of total superoxide dismutase (T-SOD) and glutathione peroxidase (GSH-PX) were assayed by using hydroxylamine method and colorimetric method respectively. The histopathological changes of the liver were observed after staining with hematoxylin-eosin (HE). RESULTS: Following modeling, the body mass and waist circumference, visceral fat weight of bilateral kidneys and testis (visceral fat weight), liver weight, serum ALT, AST, TG and TC and liver MDA contents, were significantly higher in the model group (P<0.001，P<0.05), while hepatic T-SOD and GSH-PX activity was considerably lower in the model group than those in the normal group (P<0.001). After acupuncture intervention, the levels of all the above-mentioned indexes (modeling induced both increase and decrease) were reversed relevant to the model group (P<0.05, P<0.01). The results of CT scanning showed that the fat accumulation area in the abdomen was 8.67 cm2, 18.51 cm2 and 13.75 cm2 in the normal, model and acupuncture groups, respectively, presenting a decrease after acupuncture. H．E. staining displayed that the degree of hepatic steatosis (including vague hepatic lobule boundary, disordered arrangement of hepatic cord, hepatocellular swelling, diffuse fatty degeneration, unequal-sized lipid droplets in the hepatocytes, nucleus excursion and dissolution after modeling) was improved after acupuncture. CONCLUSION: Acupuncture can reduce body weight and abdominal fat accumulation in abdominal obese NAFLD rats, which may be related to its effects in inhibiting oxidative stress (lowering MDA level and increasing the activity of T-SOD and GSH-PX) and improving hepatic lipid metabolism.

[Acupuncture Treatment of Polycystic Ovarian Syndrome Patients with Abdominal Obesity by Regulating Dai Meridian: A Randomized Controlled Clinical Trial].

OBJECTIVE: To explore the therapeutic effect of acupuncture in the treatment of polycystic ovarian syndrome (PCOS) patients with abdominal obesity by regulating glucose metabolism, insulin resistance, sex hormones, etc. METHODS: Eligible PCOS volunteers were randomly divided into treatment group(n=30) and control group(n=28). The treatment group received acupuncture therapy three times a week combined with diet control and exercise. EA (2 Hz/100 Hz, 4-8 mA) was applied to bilateral Daimai (GB 26)and Tianshu (ST 25) for 20 min/time, and bilateral Daheng (SP 15), Shenshu (BL 23), Ciliao (BL 32), Guilai (ST 29), Zulinqi (GB 41) and Waiguan (SJ 5) were stimulated with filiform needles, with the needles retained for 20 min. Patients of the control group were asked to restrict intake of dietary calories (low fat, appropriate proteins, and high carbohydrate) and aerobic exercise (>40 min/time), 3 times/week. The treatment duration lasted 12 weeks. Before and after the treatment, serum fasting insulin (FINS), fasting blood-glucose (FBG), Testosterone (T), luteinizing hormone (LH) and follicle stimulating hormone (FSH) contents were assayed using immuno-fluorometry, the homeostasis model assessment of insulin resistance (HOMA -IR) and ratio of LH/FSH were calculated, and body mass index (BMI), waist circumference (WC) and waist-hip ratio (WHR) were measured, separately. RESULTS: Comparison with their own individual pre-treatment, the levels of FINS, HOMA -IR, T, LH, LH/FSH, BMI, WC and WHR (not FBG) in both control and treatment groups were significantly decreased (P<0.05, P<0.01), and the levels of FINS, HOMA-IR and WC were significantly lower in the treatment group than in the control group (P<0.01, P<0.05). No marked differences were found between two groups in the levels of FBG, T, LH, LH/FSH, BMI and WHR (P>0.05). CONCLUSION: Acupuncture can effectively reduce FINS, HOMA-IR and WC levels in PCOS patients with abdominal obesity, down-regulating insulin resistance, which is significantly superior to simple diet control plus exercise.

[Exploration of the meridian differentiation law in polycystic ovarian syndrome of hirsutism based on data mining technology].

OBJECTIVE: To analyze the distribution characteristics of hirsutism and the law of meridian differentiation in polycystic ovarian syndrome (PCOS). METHODS: For the patients of PCOS, the modified Ferriman-Gallwey score (mF-G score) was adopted to evaluate the hirsutism distribution. The clementine data mining software was used to analyze the distribution region of hirsutism and the correlation with meridian. RESULTS: A total of 141 patients of PCOS participated in the study. The most common regions of hirsutism were in the upper lip (50.35%) and the lower abdomen (34.04%). The higher frequencies of hirsutism at two regions were at the upper lip and lower abdomen (24.82%), the upper lip and chest (12.06%) and the upper lip and thigh (11.35%). In the analysis on the correlation of the upper lip, lower abdomen and chest with other regions, hirsutism was more common in the nearby regions. In reference to the running course of meridian, the frequencies of hirsutism from high to low were the conception vessel (63.12%), the thoroughfare vessel (61.70%), the liver meridian of foot-jueyin (58.16%) and the stomach meridian of foot-yangming (57.45%). According to Miraculous Pivot: Yinyang Twenty-Five Persons, the high incidence of hirsutism (59.57%） resulted from the change of qi and blood in yangming meridian. . CONCLUSION: Blood plays the important role in the growth of body hair. Regarding the yinyang attribution, the incidence of hirsutism at the yin regions is higher than the yang regions. The occurrence of hirsutism is closely related to qi and blood changes in meridians. The conception vessel, the thoroughfare vessel, the liver meridian of foot-jueyin and yangming meridian act highly on the disease.

[Randomized Controlled Clinical Trials for Acupuncture Treatment of Abdominal Obesity].

OBJECTIVE: To observe the curative effect of acupuncture intervention in reducing visceral fat content and secretary function in abdominal obesity patients. METHODS: A total of 73 cases of abdominal obesity patients were randomly divided into acupuncture group (n = 50) and control group (n = 23) according to the randomized block design. For patients of the acupuncture group, Zhongwan (CV 12), Tianshu (ST 25), Daheng (SP 15), Daimai (GB 26), Shuidao (ST 28), Waiguan (SJ 5) and Zulinqi (GB 41) were punctured with filiform needles, followed by electroacupuncture stimulation (2 Hz/100 Hz, 4-8 mA) of bilateral ST 25 and GB 26 for 20 min, once every other day for 8 weeks. In addition, the patients were also given with health education in every session of treatment. The patients of the control group were asked to receive health education including restraining wine or liquor and salt intake, stopping smoking, increasing physical activities, regular daily life habit, etc. The abdominal fat thickness was detected using a color Doppler ultrasonography, and serum visfatin level was assayed using ELISA. Additionally, the body weight, body mass index (BMI), waist circumstance (WC) and hip circumference (HC) were determind. RESULTS: After the treatment, the subcutaneous fat thickness levels including the subcutaneous fat at the mid-point between the xyphoid and the umbilicus (S1) and the right side of the umbilicus (S2), and serum visfatin content, WC and HC in both control and acupuncture groups, visceral fat at the mid-point between the xyphoid and the umbilicus (V1) and at the right side of the um- bilicus (V2), and antero-hepatic fat (AHF), perirenal fat (PRF) , and body weight and BMI in the acupuncture group were significantly reduced in comparison with pre-treatment in the same one group ( P<0.05, P<0.001), while the ultrasonic visceral fat index [UVI, = (V1 +V2)/(SI + S2)] of the control group was markedly increased (P<0.05). The S1, V1, V2, AHF, PRF and UVI levels, and BMI, WC and HC were significantly lower in the acupuncture group than in the control group (P<0.05, P<0.001). No statistical differences were found between the two groups in the S2, body weight and serum visfatin levels (P>0.05). CONCLUSION: Acupuncture therapy can effectively reduce the visceral fat content, being better than simple health education. Both acupuncture treatment and health education can decrease serum visfatin level, regulating visceral fat's secretion.

[Effects of electroacupuncture stimulation of "Daimai" (GB 26) on body weight, blood glucose and blood lipid levels in rats with metabolism syndrome].

OBJECTIVE: To observe the therapeutic effect of electroacupuncture (EA) stimulation of "Daimai" (GB 26) on metabolic syndrome in rats. METHODS: SD rats were randomly divided into blank control group (n = 9), model group (n = 9) and EA group (n = 9). The metabolic syndrome model was established by feeding the rats with high fat and sugar forage (containing 20% lard, 10% cane sugar, 10% defatted milk powder, 2% cholest, 3% yolk powder and 55% normal forage) for 12 weeks. EA (2 Hz/100 Hz, 1-2 mA) was applied to bilateral "Daimai" (GB 26) for 20 min, once daily for 4 weeks. Fasting blood glucose (FBG), serum triglyceride (TG), total cholesterol (TC), high density lipoprotein-cholesterol (HDL-C), lower density lipoprotein-cholesterol (LDL-C) contents were detected using chemical methods,fasting serum insulin (FINS) was determined by using radioimmunoassay. Homeostasis model assessment-index of resistance (HOMA-IR, insulin resistance index = FBG x FINS/22.5) was calculated. RESULTS: Compared to the blank control group, the body weight (from the 8th week on),abdominal circumference (from the 4th week on), and FBG, FINS, TG, TC, LDL-C contents as well as HOMA-IR in the model group were significantly increased (P < 0.05, P < 0.01), while the HDL-C content was considerably lowered after modeling (P < 0.05). In comparison with the model group, the body weight on the 16th week, FBG, FINS, HOMA-IR, TG, TC and LDL-C levels were evidently down-regulated in the EA group (P < 0.05, P < 0.01). No significant differences were found between the model and EA groups in the abdominal circumference and HDL-C content (P > 0.05). CONCLUSION: Electroacupuncture stimulation of "Daimai" (GB 26) can reduce insulin resistance, regulate blood lipid, and lower blood glucose in metabolic syndrome rats.

[Acupuncture treatment of abdominal obesity patients by "belt vessel (Daimai) regulating method"].

OBJECTIVE: To observe the clinical effect of acupuncture intervention on abdominal obesity by stimulation of "Belt Vessel (Daimai) Regulation Acupoint Recipe". METHODS: A total of 35 abdominal obesity outpatients were recuited in the present study. The "Belt Vessel (Daimai) Regulation Acupoint Recipe" was composed of Daimai (GB 26), Tianshu (ST25), Daheng (SP 15), Zhongwan (CV 12), Liangmen (ST 21), Shuidao (ST 28), Zusanli (ST 36), Shangjuxu (ST 37), Fenglong (ST 40) and Zulinqi (GB 41, both sides except CV 12). In addition, electroacupuncture was also applied to bilateral GB 26 and ST 25. The treatment was conducted once every other day, 8 weeks altogether. The body weight, body mass index (BMI), waist circumference (WC), hip circumference (HC), Waist-to-hip ratio (WHR), waist-to-height ratio (WHtR) and body fat rate [ = 1.2 x BMI + 0.23 x age-5.4-10.8 x sex (male = 1, female = 0)] were measured or calculated. The thickness of abdominal adipose layer (i.e.; S1 = distance from the interface of the skin and subcutaneous fat to the linea alba abdominis at the midpoint between the xiphoid-process and the umbilicus; S2 = distance from the interface of the skin and subcutaneous fat to the anterior bounder of the external oblique muscle of abdomen on the right side closely to the umbilicus center) and visceral fat layer thickness (V 1 = distance from the midpoint of the linea alba abdominis to the anterior bounder of the vertebra body; V 2 = distance from the peritoneum to the right bounder of the vertebra body at the umbilicus level) were measured for calculating ultrasound viscerofatty index [UVI = (V 1 + V2)/(S 1 + S 2)] by using a color Doppler ultrasonography. RESULTS: After 8 weeks' treatment, the body weight, BMI, waist and hips circumferences, WHtR and percentage of body fat were declined significantly (P < 0.05, P < 0.001). The WHR was decreased compared to that of pre-treatment, but without statistical significance (P > 0.05). The thickness of abdominal subcutaneous adipose and visceral adipose layers and UVI were decreased significantly following the treatment (P < 0.05, P < 0.001). The reduction of visceral adipose layer was superior to that of the abdominal subcutaneous one (P < 0.001). CONCLUSION: Acupuncture intervention has a significant effect on reducing abdominal obesity in abdominal obesity patients.