[Effect of acupoint thread-embedding on macrophage polarization of epididymis adipose tissue in obese mice].

OBJECTIVE: To observe the effect of acupoint thread-embedding at "Zusanli" (ST 36) and "Fenglong" (ST 40) on the macrophage polarization of epididymis adipose tissue in obese mice, and to explore the action mechanism of acupoint thread-embedding on weight control. METHODS: Among 30 male C57BL/6 mice, 10 mice were randomly selected and fed with normal diet, and the remaining 20 mice were fed with high-fat diet to establish the obesity model. Sixteen mice with successful obesity model were randomly divided into a model group and an acupoint thread-embedding group, 8 mice in each group. Eight mice were selected from mice which were fed with normal diet as the normal group. On the next day of successful modeling, acupoint thread-embedding was performed at "Zusanli" (ST 36) and "Fenglong" (ST 40) in the acupoint thread-embedding group, once every 10 days for 4 times. The body weight was recorded at 0, 8, 16, 24, 32, 40 days into intervention; the level of glucose metabolism was compared after intervention; the level of lipid metabolism and weight of epididymal adipose tissue were compared at the end of the intervention; the mRNA expression of M1 and M2 macrophage-related cytokines interleukin-10 (IL-6), monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) were detected by real-time PCR; the mRNA and protein expression of M1 macrophage labeled inducible nitric oxide synthase (iNOS) and M2 macrophage labeled arginase-1 (Arg-1) were detected by real-time PCR and Western blot. RESULTS: Compared with the normal group, the body weight at 0, 8, 16, 24, 32, 40 days into intervention in the model group was increased (P<0.05); the results of glucose tolerance test at 0, 30, 60, 120 min and insulin tolerance test at 0, 30, 60, 90, 120 min in the model group were higher than those in the normal group (P<0.05); the levels of total cholesterol and triacylglycerol in the model group were significantly higher than those in the normal group (P<0.001, P<0.01); the weight of epididymal adipose tissue in the model group was significantly higher than that in the normal group (P<0.001); the mRNA expression of IL-6, MCP-1, TNF-α and iNOS was increased (P<0.05, P<0.01, P<0.001), that of IL-10, Arg-1 was decreased (P<0.01), the protein expression of iNOS was up-regulated (P<0.01), and that of Arg-1 was down-regulated (P<0.001). Compared with the model group, the body weight at 16, 24, 32, 40 days into treatment in the acupoint thread-embedding group was reduced (P<0.05); the results of glucose tolerance test at 30, 60, 120 min and insulin tolerance test at 30, 60 min in the acupoint thread-embedding group were lower than those in the model group (P<0.05); the levels of total cholesterol and triacylglycerol in the acupoint thread-embedding group were significantly lower than those in the model group (P<0.01, P<0.05); the weight of epididymal adipose tissue in the acupoint thread-embedding group was significantly lower than that in the model group (P<0.01); the mRNA expression of IL-6, MCP-1, TNF-α and iNOS was reduced (P<0.05), that of IL-10, Arg-1 was increased (P<0.05), the protein expression of iNOS was down-regulated (P<0.05), and that of Arg-1 was up-regulated (P<0.01). CONCLUSION: Acupoint thread-embedding at "Zusanli" (ST 36) and "Fenglong" (ST 40) may play a role in weight control by regulating the polarization of macrophages.

[Effect of acupuncture on oxidative stress and apoptosis-related proteins in obese mice induced by high-fat diet].

OBJECTIVE: To explore the effect of acupuncture on oxidative stress and apoptosis-related proteins of liver in obese mice induced by high-fat diet. METHODS: A total of 45 male C57BL/6 mice were randomized into a control group (10 mice) and a model established group (35 mice). Mice in the model established group were fed with high-fat diet for 16 weeks to establish the obesity model. After model established, 30 mice were randomized into a model group, a non-acupoint group and an acupoint group, 10 mice in each one. Acupuncture was applied at "Guanyuan" (CV 4), " Zusanli" (ST 36), "Yishu" (EX-B 3) in the acupoint group and the points of 0.5 cm and 1 cm to the base of tail in the non-acupoint group, 15 min each time, once a day for 8 weeks. Mice in the control group were fed with normal diet, while mice in the other 3 groups were fed with high-fat diet continuously for 8 weeks. The body weight was measured at 0, 4th, 8th, 12th, 16th, 20th, 24th week in each group respectively. After 24-week intervention, the weight of white adipose tissue of epididymis and perirenal and liver was measured; the levels of serum alanine transaminase(ALT) and glutamic oxaloacetic aminotransferase (AST) were detected by automatic biochemical analyzer; liver homogenate was used to detect the level of malondialdehyde (MDA) and the activity of superoxide dismutase (SOD); the liver morphology was observed by HE staining; the expression of apoptosis-related proteins Bax and Bcl-2 were detected by Western blot. RESULTS: Compared with the control group, the body weight of mice in the model group, the acupoint group and the non-acupoint group was decreased on 16th week into experiment (before intervention, P<0.05); compared with the model group and the non-acupoint group, the body weight of mice in the acupoint group were decreased after intervention (P<0.05). Compared with the control group, the weight of white adipose tissue and liver, the levels of serum ALT and AST, the level of liver MDA, the expression of liver Bax were increased (P<0.05); the activity of liver SOD and the expression of liver Bcl-2 were decreased in the model group after intervention (P<0.05). Compared with the model group and the non-acupoint group, the weight of white adipose tissue and liver, the levels of serum ALT and AST, the level of liver MDA, the expression of liver Bax were decreased (P<0.05); the activity of liver SOD and the expression of liver Bcl-2 were increased in the acupoint group after intervention (P<0.05). CONCLUSION: Acupuncture at "Guanyuan" (CV 4), "Zusanli" (ST 36) and "Yishu"(EX-B 3) can improve obesity and obesity related hepatic disorder by regulating oxidative stress and inhibiting apoptosis in liver.

Ehrlichia species in pond-farmed leeches (Hirudinaria sp.) in Hubei Province, China.

Leeches are frequently used in traditional Chinese medicine. However, they are potentially hazardous to human and animal health by transmitting several pathogens. Studies of diseases transmitted by leeches are scarce. The purpose of this study was to analyze the pathogens carried in pond-farmed medicinal leech in China. Leeches were collected from 6 farms in Hubei Province in central China. DNA was extracted from the internal organ of leeches to analyze the origin of blood meal. Leech genera were confirmed through amplification of 18S rRNA and mitochondrial gene cytochrome oxidase I (COI) gene by PCR and host animal species were identified through amplification of mitochondrial cytochrome b gene. Species of Ehrlichia in the leech specimens were screened with PCR using specific primers. PCR amplification and DNA sequencing showed that 620 leeches were Hirudinaria sp. Ehrlichia DNA was detected in 39 specimens from 2 farms. We obtained a total of 65 sequences of the cytB gene from 620 leech internal organ samples including sequences of human (n = 5), rat (n = 1), domestic pig (n = 10), duck (n = 23), goose (n = 12) and buffalo (n = 14). Phylogenetic analysis of the rrs and groEL gene sequences showed that Ehrlichia detected in the study were closely related to Ehrlichia sp. in ticks from Korea and Japan. To the best of our knowledge, this is the first report on Ehrlichia DNA being detected from leeches. Our findings provided new data on Ehrlichia spp. and farmed leech species in China.