Chinese Archives of Traditional Chinese Medicine,Part 1: Mechanism Study,Vol 35,No. 01
Objective: To study the effects of Bushen Qutan Formula on leptin, Ob-Rb, TNFα, INFγ, PGE
2, MMP-9, MMP-13 and MMP-1 mRNA expression levels in obese rats with osteoarthritis (OA) due to kidney-deficiency by PCR. Method: The rats were dividied into normal group, model group, low-dose, middle-dose and high-dose Bushen Qutan Formula groups, and glucosamine sulfate capsule suspension group. Rats in the normal group and model group were administered with normal saline by gavage each day and those in the various treatment groups were administered with the corresponding doses for seven continuous days by gavage. Rats were killed after the last drug administration. In the bacteria-free environment, we decalcified the cartilage and studied the levels of leptin, Ob-Rb, TNFα, INFγ, PGE
2, MMP-9, MMP-13 and MMP-1 mRNA expression in rats by PCR after obtaining of the cartilage. Result: Compared with the normal group, there was no statistical significances in the mRNA expression levels of Ob-Rb, TNFα, INFγ, PGE
2, MMP-9 and MMP-1 after the intervention of Bushen Qutan Formula, while there were significantly statistical significances in the mRNA expression levels of leptin and MMP-13. The mRNA expression levels of leptin and MMP-13 in the model group were increased (
P < 0.01). Compared with the model group, the mRNA expression levels of leptin and MMP-13 in low-dose, middle-dose, high-dose Bushen Qutan Formula groups and glucosamine sulfate capsule suspension group were decreased (
P < 0.05), especially in the high-dose Bushen Qutan Formula group. Conclusion: The obese rat model with OA due to kidney-deficiency was stable, which would provide the new method for modeling. Bushen Qutan Formula could decrease the mRNA expression levels of leptin and MMP-13 in obese rats with OA due to kidney-deficiency, which would provide the theoretical basis for clinical study of Bushen Qutan Formula.
World Chinese Medicine,Part 1: Mechanism Study,Vol 14,No. 02
Objective: To observe the effects of ginsenoside Rb1 on body weight, body composition, blood lipid, skeletal muscle endurance and insulin sensitivity in obese mice, and further explore its effects on AMPK signaling pathway in skeletal muscle. Methods: The 8-week-old C57BL/6J mice were fed with high fat diet for 12 weeks to induce obese mice model. The obese mice were randomly divided into three groups (the model control group, the metformin group and the ginsenoside Rb1 group). The peer mice fed with normal diet were assigned into the normal control group. The mice were administrated with the corresponding drugs for eight weeks. Body weight and food intake of the mice were measured regularly every week. Treadmill test was performed in the 3 rd and 7 th weeks, and the oral glucose tolerance test was conducted in the 4 th and 8 th weeks. Body composition of the mice was detected by NMR Animal Body Composition Analyzer on the 8th week. Four indexes of plasma lipids and free fatty acid level were detected at the end of the study. The mRNA expressions of AMPKα and PGC-1α in skeletal muscle were examined by real-time fluorescence quantitative PCR (FQPCR), and the protein expressions of AMPKα, p-AMPKα and PGC-1α were detected by Western blot. Results: The body weight (since the 5 th week) and food intake of the mice in the ginsenoside Rb1 group were significantly lower than those in the model control group (
P < 0.05) in a relatively time-dependent manner. Ginsenoside Rb1 could significantly reduce the levels of triglyceride and low density lipoprotein cholesterol, and increase the high density lipoprotein cholesterol (
P < 0.05). In addition, ginsenoside Rb1 reduced serum free fatty acid level (P < 0.05). After the administration of ginsenoside Rb1 for eight weeks, the body fat mass of obese mice was decreased and the lean mass was increased, oppositely (
P < 0.05). The skeletal muscle endurance (
P < 0.05) and the oral glucose tolerance of the obese mice were improved by ginsenoside Rb1. At the molecular level, ginsenoside Rb1 could up-regulate the mRNA and protein expression of AMPKα in skeletal muscles, and the content of p-AMPK protein was also increased significantly (
P < 0.05). At the same time, the mRNA and protein expression levels of PGC-1α were also up-regulated, correspondingly (
P < 0.01). Conclusion: Ginsenoside Rb1 exerts effects on reducing body weight, decreasing blood lipid levels, improving skeletal muscle endurance and insulin sensitivity in obese mice by activating AMPK signaling pathway in skeletal muscles.
Mycosystema,Part 1: Mechanism Study,Vol 38,No. 03
This study aims at screening the active extract from
Isaria cicadae for controlling obesity and its complications. An obesity mouse model was prepared by feeding high fat and cholesterol diet to mice.
Isaria cicadae extracts (ethyl alcohol extract, aqua destillata extract and alkaline water extract) were fed to the mice for 35 days by gavage. Histopathological changes of liver and colon, serum levels of total cholesterol (T-CHO), total triglycerides (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoproteincholesterol (HDL-C) and gut microbiota were monitored. Results showed that all of the three extractions could reduce the body weight and improve the serum levels of T-CHO, TG, LDL-C and HDL-C of the experimental mice. The pathological observation revealed that the damages of liver and colon tissues induced by high fat and cholesterol diet were ameliorated more or less, and the alpha and beta diversity and gut microbiota composition were also improved. The ethyl alcohol extract of
Isaria cicadae (ICE) has the best effect among the three tested samples, indicating that ICE would be the most active extract for obesity control while the functional mechanism remains to be studied.
Acupuncture Research,Part 1: Mechanism Study,Vol 40,No. 04
Objective To observe the effect of electroacupuncture(EA)stimulation of“Housanli”(ST 36) and “Zhongwan”(CV 12) on serum leptin level and expression of phosphorylated Janus kinase 2(p-JAK2)and phosphorylated signal transducer and activator of transcription 3(p-STAT3)proteins in the liver tissue of obese rats.Methods Thirty-two male Wistar rats were randomly divided into control group,model group,manual acupuncture(MA)group and EA group(n=8 in each group).The obese model was induced by high fat diet for 8 weeks.MA or EA(20Hz,1.5V)stimulation was applied to bilateral“Housanli”(ST 36) and “Zhongwan”(CV 12) for 20 min,once daily for 4weeks.The animals' body weight and length were recorded,and Lee's index was calculated.Serum leptin level and liver p-JAK2 and p-STAT3 protein expression levels were detected using radioimmunoassay and Western blot,respectively.Results In comparison with the control group,the rats' body weight,Lee's index,and serum leptin content were remarkably increased in the model group(P<0.05),while hepatocellular p-JAK 2 and p-STAT3 protein expression levels were significantly down-regulated in the model group(P<0.05).After treatment for 4 weeks,the levels of body weight,Lee's index,serum leptin were significantly reduced(P<0.05),and hepatocellular p-JAK 2 and p-STAT 3 protein expression in the MA and EA groups were considerably up-regulated compared with the model group(P<0.05).The effect of EA was apparently superior to that of MA in up-regulating p-JAK 2 and p-STAT 3protein expression(P<0.05).Conclusion Both EA and MA stimulation can reduce the obese rats' body weight,and Lee's index,which may be closely associated with their effects in down-regulating serum leptin and in enhancing hepatocellular p-JAK 2and p-STAT 3protein expression.
Chinese Journal of Integrated Traditional and Western Medicine,Part 1: Mechanism Study,Vol 36,No. 05
Objective To explore the mechanism of Electroacupuncture (EA) for reducing diet-induced obesity (DIO) rat weight through tuberous sclerosis complex 1 (TSC1)-mammalian target of rapamycin (m TOR) signal pathway in hypothalamus. Methods Forty male SD rats were randomly divided into the model group (
n = 30) and the normal control group (
n = 10). DIO rat model was prepared by high fat forage for 12 successive weeks. Successfully modeled 19 rats were further randomly divided into the model group (
n = 9) and the EA group (
n = 10). EA at Tianshu (ST25), Sanyinjiao (SP6), Zhongwan (RN12), Zusanli (ST36) was performed in the EA group, 5 successive days per week with a 2-day rest, 4 weeks in total. No intervention was given to rats in the model group and the normal control group. Body weight was observed in all rats. Methylation of TSC1 promotor was detected by bisulfite sequencing method. m RNA expression of m TOR in hypothalamus was detected by RT-PCR. Results After EA treatment body weight in the EA group was obviously reduced (
P < 0. 05). Compared with the normal control group, body weight was not statistically different between the model group and the EA group after treatment (
P > 0. 05). Methylation rate of TSC1 promotor was higher in model group (94. 0% ± 4. 5%) than in the normal control group (87. 0% ± 3. 6%) and the EA group (87. 4% ± 3. 9%) (
P < 0. 05). Expression of m TOR in the model group (1. 84 ± 0. 51) was higher than that in the normal control group (1. 02 ± 0. 22) and the EA group (1. 46 ± 0. 29) (
P < 0. 05). Conclusion EA could lower DIO rats’ body weight by down-regulating methylation rate of TSC1 promotor and regulating expression of m TOR in hypothalamus.
Acupuncture Research,Part 1: Mechanism Study,Vol 42,No. 06
Objective To investigate the effects of electroacupuncture (EA) on the expressions of adenosine receptor (AR) in the white adipose tissue (WAT) of diet-induced obese (DIO) mice, so as to reveal a peripheral mechanism of EA underlying improvement of body weight. Methods Forty three-week-old C 57BL/6 male mice were divided into normal diet group (
n = 12) and high fat diet group (
n = 28) randomly, and fed by normal diet and high fat diet for 12 weeks, respectively. In the high fat diet group, mice with body weight over 20% heavier than that of the normal diet group were considered as obese mice. The normal diet mice and the obese mice were divided into common diet group (CD,
n = 5) and common diet with eletroacupuncture group (CD + EA,
n = 7), or high fat diet group (HFD,
n = 6) and high fat diet with eletroacupuncture group (HFD + EA,
n = 12). The CD + EA group and the HFD + EA group were treated with EA at “Housanli” (ST 36) and “Neiting” (ST 44), [2 Hz/15 Hz, (0.6–1.0) mA] for 20 min, 6 times a week for 4 weeks. Body weight, ratio of WAT/body weight were calculated, and qPCR and Western blot were applied to detect mRNA and protein levels of adenosine receptors in the epididymal adipose tissue (Epi-WAT), respectively. Results Compared with the normal diet group, high fat diet significantly increased body weight in C 57BL/6 mice after feeding for 12 weeks (
P < 0.01); 18 out of 28 mice in the high fat diet group were considered as obesity. After treatment, the body weight and the ratio of Epi-WAT / body weight of the HFD group were increased compared with those in CD group (
P < 0.05), the changes of body weight in HFD group were bigger than that in CD group (
P < 0.01). Compared with HFD group, the body weight and the ratio of Epi-WAT/ body weight of HFD + EA group were decreased after EA (
P < 0.05), the changes of body weight were also significantly increased (
P < 0.01). No significant differences were found among the four groups in the expression levels of A
1R mRNA (
P > 0.05). The expression of A
3R mRNA in HFD group was lower than that in CD group (
P < 0.01), while the expressions of A
2AR and A
2BR proteins were decreased in HFD group compared with those in CD group (
P < 0.01). In comparison with HFD group, the expression levels of A
2AR and A
2BR mRNAs and proteins were significantly up-regulated in HFD + EA group, respectively (
P < 0.05,
P < 0.01). Conclusions EA intervention was able to reduce the body weight of DIO mice, which might be associated with its effects in regulating the expressions of A
2AR and A
2BR in WAT, suggesting a new mechanism of EA in accelerating peripheral WAT metabolism.
Acupuncture Research,Part 1: Mechanism Study,Vol 43,No. 07
Objective To observe the effect of electroacupuncture (EA) on the activities of peroxisome proliferator-activated receptor gamma coactivator-1 alpha/uncoupling protein-1 (PGC-1α/UCP-1) signaling pathway in white adipose tissue (WAT) of diet-induced obesity (DIO) rats, so as to reveal its underlying mechanism in body weight loss. Methods Thirty-four male Wistar rats were randomly divided into normal diet (
n = 10), high fat diet (model), sham EA-acupoint and EA groups (
n = 8 in each of the latter three groups). The obesity model was established by feeding the rats with high fat diet containing lard oil, sugar, albumen powder, cholesterol, salt and sodium cholate for 12 weeks. EA (2 Hz/15 Hz, 1 mA) was applied to bilateral “Zusanli” (ST 36) and “Tianshu” (ST 25) or sham acupoints [about 5 mm lateral to Zusanli (ST 36) and Tianshu (ST 25)] for 30 min, once daily, five times per week for a total of eight weeks. During the treatment, all rats were fed with normal diet, and their body weight and length were measured once a week for calculating the Lee’s index. The contents of serum total cholesterol (TC) and triglyceride (TG) were measured by using biochemical methods. The immunoactivity of PGC-1α and UCP-1 in the abdominal WAT was detected by immunohistochemistry. Results After modeling, the Lee’s index, serum TC and TG contents were significantly increased, and the levels of serum HDL-C, and PGC-1α and UCP-1 immunoactivity in WAT were considerably decreased in the model group relevant to those the control group (
P < 0.05). Following the treatment, the Lee’s index, TC and TG contents were significantly down-regulated while HDL-C and PGC-1α and UCP-1 immunoactivity were obviously up-regulated in the EA-acupoint group relevant to those in the model group (
P < 0.05). Conclusion EA can effectively reduce the body weight and adipose content in obesity rats, which may be closely related to its efficacy in up-regulating PGC-1α/UCP-1 signaling in WAT, suggesting the efficacy of EA in promoting the browning of WAT.
Acupuncture Research,Part 1: Mechanism Study,Vol 44,No. 08
Objective To investigate the effect of electroacupuncture (EA) plus treadmill exercise on the expression of peroxisome proliferator activated receptor γ coactivator 1α (PGC-1α), irisin, AMP-activated protein kinase (AMPK) in skeletal muscle of diet-induced obesity (DIO) rats, so as to explore its mechanism underlying body reduction promotion.
Methods Forty-two male SD rats were divided into normal diet (control,
n = 10), high-fat diet (model), EA, treadmill exercise and EA plus treadmill exercise (combination) groups (
n = 8 in each of the latter four groups). The obesity model was established by feeding the rats with high fat diet. EA (2 Hz/15 Hz, 1 mA) was applied to bilateral “Zusanli (ST 36)” and “Tianshu (ST 25)” for 30 min, five times per week for a total of eight weeks. Rats of the treadmill exercise group were forced to perform exercise on a treadmill (16 m/min) for 30 min, five times per week for a total of eight weeks. Rats in the combination group received the above-mentioned two methods. During the treatment, rats in the control group were fed with normal fodder, while rats in the other groups were fed with high fat fodder. Their body weight was measured once a week. The mRNA and protein expression levels of PGC-1α, fibronectin type Ⅲ domain containing 5 (FNDC5), and AMPK in the skeletal muscle were measured by quantitative real-time PCR and Western blot assay, respectively.
Results After modeling, the body weight was significantly increased (
P < 0.05), and the mRNA and protein expression levels of PGC-1α and FNDC5, as well as AMPK mRNA expression and phosphorylation level in the skeletal muscle were considerably decreased in the model group relevant to those in the control group (
P < 0.05). Following the treatment, the body weight was significantly down-regulated, while the mRNA and protein expression levels of PGC-1α and FNDC5, as well as AMPK mRNA expression and phosphorylation level were obviously up-regulated in the EA, treadmill exercise and combination groups relevant to those in the model group (
P < 0.05). The therapeutic effect of EA plus treadmill exercise was significantly superior to those of both simple EA and simple treadmill exercise in down-regulating the body weight, as well as in up-regulating the mRNA and protein expression levels of PGC-1α and FNDC5, as well as AMPK mRNA expression and phosphorylation level (
P < 0.05).
Conclusion Both EA and treadmill exercise can significantly increase the expression of PGC-1α, FNDC5 and p-AMPK in skeletal muscle of DIO rats, suggesting their efficacy in restoring fatty acid oxidation in skeletal muscle cells and improving mitochondrial function, which may contribute to body reduction. The therapeutic effect of EA plus treadmill exercise is better than those of simple EA and simple treadmill exercise.
Chinese Acupuncture & Moxibustion,Part 1: Mechanism Study,Vol 39,No. 09
Objective To observe the effects of electroacupuncture (EA) on inflammatory response and intestinal mucosal barrier in obese rats with insulin resistance (IR), and to explore the mechanism of EA in improving IR in rats.
Methods Among 45 Wistar male rats, 15 rats were randomly selected and fed with common diet. After eight weeks, 10 rats were randomly selected as the normal control. The remaining 30 rats were fed with high-fat diet for eight weeks to induce the obesity model. Among the 28 successfully modeled rats, 20 were randomly selected and divided into the model group and EA group, with 10 rats in each group. At the same time, three rats in the model group and the EA group were randomly selected for hyperinsulinemia euglycemic clamp operation to determine whether the IR modeling was successful. After model establishment, the rats in the EA group were intervened with EA at “Zhongwan” (CV 12), “Guanyuan” (CV 4), “Zusanli” (ST 36) and “Fenglong” (ST 40) with continuous wave at the frequency of 2 Hz and intensity of 1 mA. The bilateral “Zusanli” (ST 36) and “Fenglong” (ST 40) were used alternately. The needles were retained for 10 min. EA was given once every other day, three times a week for a total of eight weeks. During the intervention, the rats in the normal group and the model group were fixed without receiving any intervention. The body mass and postprandial blood sugar were measured in each group before and after 8-week intervention. After six-week intervention, three rats in each group were clamped to detect systemic insulin sensitivity. Before the rats were sacrificed, the blood was taken from the apex of the heart to detect the serum insulin content. After the sacrifice, the mRNA levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in liver and adipose tissue and occlidin and ZO-1 in colon tissue were detected by real time-PCR assay. The protein expression levels of occludin and ZO-1 in colon tissue were detected by Western blot method.
Results Compared with the normal group, the body mass, postprandial blood sugar and serum insulin content in the model group were increased significantly, and the glucose infusion rate (GIR) was decreased significantly (all
P < 0.01). The mRNA expressions of TNF-a and IL-6 in liver and adipose tissue were increased (
P < 0.05,
P < 0.01). The mRNA and protein expressions of ZO-1 in colon tissue were decreased significantly (both
P < 0.01), and the mRNA expression of occludin was significantly decreased (
P < 0.01). Compared with the model group, the body mass, postprandial blood sugar, serum insulin content, mRNA expressions of TNF-a and IL-6 in liver and adipose tissue were significantly decreased (
P < 0.01,
P < 0.05), while GIR was significantly increased (
P < 0.01). The mRNA and protein expressions of ZO-1 in colon tissue were increased (
P < 0.01,
P < 0.05).
Conclusion EA could decrease blood sugar and increase insulin sensitivity, which might be related to its efficacy in inhibiting the expression levels of inflammatory factors and improving intestinal mucosal barrier.
Acupuncture Research,Part 1: Mechanism Study,Vol 44,No. 10
Objective To observe the effect of electroacupuncture (EA) on inflammatory reaction and insulin sensitivity in insulin-resistant obese (OIR) rats.
Methods Thirteen male Wistar rats were randomly selected as the control group and fed with common diet. The other 39 rats were fed with high-fat diet for eight weeks to establish OIR model and then randomized into model, EA and sham EA groups. EA (2 Hz, 1 mA) was applied to unilateral Zusanli (ST36), Fenglong (ST40), Zhongwan (CV12) and Guanyuan (CV4) for 15 min, once every other day for eight weeks, and sham EA was applied to unilateral four control spots about 5 mm lateral to the aforementioned four acupoints after shallowly inserting acupuncture needles, but without electric current output. After eight weeks’ intervention, the body weight was recorded and the glucose infusion rate (GIR) measured using hyperinsulinemic-euglycemic clamp technique. At the 6
th week of intervention, glucose contents of intraperitoneal glucose tolerance test (IPGTT) and intraperitoneal insulin tolerance test (IPITT) were measured. The levels of serum insulin (INS) and inflammatory factors as C-reactive protein (CRP), IL-6 and TNF-α were measured by using ELISA at the end of the treatment. The expression levels of IL-6, TNF-α, monocyte chemoattractant protein-1 (MCP-1), IL-10 and IL-1β proteins and mRNAs in the abdominal adipose tissues were detected by Western blot and quantitative real time-PCR, separately. The CD68 expression (displaying infiltration of macrophages) of adipose tissue was detected using immunohistochemistry.
Results After modeling, the contents of glucose of IPGTT at 30, 60 and 120 min and those of IPITT at 15, 30, 60 and 120 min, serum INS, CRP, IL-6 and TNF-α, as well as the expression levels of IL-6, TNF-α, IL-1β and MCP-1 proteins and mRNAs and CD68 protein were significantly increased (
P < 0.01,
P < 0.05), while the levels of GIR and IL-10 protein and mRNA were obviously decreased in the model group in comparison with those of the control group (
P < 0.01), suggesting an increase of inflammation and a decline of INS sensitivity. Following the interventions, the increased contents of glucose of IPGTT and IPITT, serum INS, CRP, IL-6 and TNF-α, expression levels of IL-6, TNF-α, IL-1β and MCP-1 proteins and mRNAs and CD68 protein, and the decreased levels of GIR and IL-10 protein and mRNA were evidently reversed in the EA group compared with the model group (
P < 0.01,
P < 0.05) rather than those in the sham EA group (
P > 0.05).
Conclusion EA can reduce the level of inflammation and improve insulin sensitivity in OIR rats.
