ABSTRACT
OBJECTIVES: To investigate the effect of electroacupuncture(EA) at "Changbing Decoction" on alleviating ulcerative colitis (UC) and regulating the polarization of colonic macrophages in rats, so as to explore its mechanisms underlying improvement of UC. METHODS: Twenty-six male SD rats were randomly divided into 4 groupsï¼normal group(6 rats), model group(8 rats), EA group(6 rats), and western medication group(6 rats). The rat model of UC was established by using 5% dextran sulfate sodium (DSS) solution drinking water for 7 days, followed by drinking 1% DSS solution during treatment period. After 7-day model establishment, EA treatment(10 Hz/50 Hz, 20 min) was applied to "Zhongwan"(CV12), bilateral "Tianshu"(ST25) and "Shangjuxu"(ST37) for 3 d, and rats in the western medication group were given mesalazine suspension(200 mg/kg) by gavage for 3 d. The body weight, spleen weight and colon length of rats were measured. The disease activity index (DAI) score was evaluated. The morphological changes and inflammatory cell infiltration of colon were detected after HE staining and pathological scores were eva-luated. The contents of tumor necrosis factor α(TNF-α), interleukin(IL)-1ß, IL-2 and IL-10 in serum were detected by ELISA. The protein expressions of M1 and M2 macrophage markers nitric oxide synthase (iNOS) and arginase 1(Arg1) were detected by fluorescence double staining and Western blot, respectively. Quantitative real-time PCR was used to detect iNOS and Arg1 mRNA expressions. RESULTS: Compared with the normal group, rats in the model group had increased pathological damage degree and inflammatory cell infiltration in the colon tissue, slowed-down body weight gain, decreased colon length, spleen weight, serum anti-inflammatory factors IL-2 and IL-10 contents, colonic Arg1/CD68 fluorescence positive expression, and Arg1 protein and mRNA expressions(P<0.01, P<0.05), as well as increased DAI scores, colon histopathological scores, contents of serum pro-inflammatory factors TNF-α and IL-1ß, colonic iNOS/CD68 fluorescence positive expression, iNOS protein and mRNA expressions(P<0.01). Compared with the model group, the above indicators were significantly improved in rats of the EA group and the western medication group(P<0.01, P<0.05). CONCLUSIONS: EA of "Changbing Decoction" can improve UC of rats by regulating the polarization of colonic macrophages, inhibiting the generation of M1 macrophages and promoting the generation of M2 macrophages.
Subject(s)
Colitis, Ulcerative , Electroacupuncture , Rats , Male , Animals , Colitis, Ulcerative/genetics , Colitis, Ulcerative/therapy , Interleukin-10/genetics , Rats, Sprague-Dawley , Tumor Necrosis Factor-alpha/genetics , Interleukin-2 , Macrophages , RNA, Messenger , Body WeightABSTRACT
OBJECTIVE: To observe the effect of electroacupuncture ï¼EAï¼ at "Zhongwan" ï¼CV12ï¼, "Tianshu" ï¼ST25ï¼ and "Shangjuxu" ï¼ST37ï¼ ï¼an acupoint prescription "Changbingfang" for treatment of intestinal disordersï¼ on autophagy and expression of AMPK/mTOR signaling pathway in rats with ulcerative colitis ï¼UCï¼, so as to explore its mechanism underlying improvement of UC. METHODS: Thirty-two male SD rats were randomly divided into control, model, medication and EA groups, with 8 rats in each group. The UC model was established by free drinking of 5% dextran sulfate sodium salt solution for 7 days. EA stimulation ï¼10 Hz/50 Hzï¼ was delivered to CV12, ST25 and ST37 for 20 min, once a day for 3 consecutive days. Rats of the medication group received gavage of mesalazine suspension ï¼200 mg/kgï¼ once a day, 3 times in total. The rats' general conditions were recorded for calculating the disease activity index ï¼DAIï¼ score ï¼0-4 pointsï¼. Histomorphological changes of colon were observed via HE staining. The levels of serum interleukin 6 ï¼IL-6ï¼, tumor necrosis factor-α ï¼TNF-αï¼ and IL-10 were measured by ELISA. The mRNA expressions of LC3B and p62 were tested by fluorescence quantitative PCR. Western blot was used to detect the expression levels of LC3B, p62 and AMPK/mTOR pathway related proteins in colon tissues. RESULTS: Compared with the control group, the DAI score, contents of serum IL-6 and TNF-α, the expression levels of p62 protein and mRNA, ratio of p-mTOR/mTOR were significantly increased ï¼P<0.01ï¼ï¼ while the content of serum IL-10, the expression levels of LC3B mRNA, ratio of LC3Bâ ¡/LC3Bâ and p-AMPK/AMPK were decreased ï¼P<0.01, P<0.05ï¼ in the model group. Relevant to the model group, modeling-induced increases of DAI score, serum IL-6, TNF-α and IL-10 contents, expressions of p62 protein and mRNA, LC3B mRNA, ratio of p-mTOR/mTOR, LC3Bâ ¡/LC3Bâ and p-AMPK/AMPK were reversed in both medication and EA groups ï¼P<0.01, P<0.05ï¼. The effect of EA was apparently superior to that of mesalazine in up-regulating ratio of LC3Bâ ¡/LC3Bâ and p-AMPK/AMPK, p62 mRNA expression ï¼P<0.01, P<0.05ï¼, and in down-regulating ratio of p-mTOR/mTOR ï¼P<0.05ï¼. H.E. staining showed severe damage of the colonic mucosal barrier with infiltration of a large number of inflammatory cells in the model group, which was milder in medication and EA groups. CONCLUSION: EA of acupoint recipe "Changbingfang" can improve the symptoms in UC rats, which may be related to its functions in promoting colonic autophagy, increasing AMPK phosphorylation level, and decreasing mTOR phosphorylation level.
Subject(s)
Colitis, Ulcerative , Electroacupuncture , Male , Animals , Rats , Rats, Sprague-Dawley , Colitis, Ulcerative/genetics , Colitis, Ulcerative/therapy , AMP-Activated Protein Kinases/genetics , Interleukin-10 , Mesalamine , Interleukin-6 , Tumor Necrosis Factor-alpha/genetics , Signal Transduction , TOR Serine-Threonine Kinases/genetics , RNA, Messenger , AutophagyABSTRACT
In order to study the influencing factors on Raman spectroscopy, we research a series of comparative Raman spectroscopy of multi-walled carbon nanotubes (MWCNT) with different tube diameter and length. The results suggest that the G peak and D peak of MWCNT are all red-shifted as compared to that of polycrystalline graphite; In the same conditions, the peak intensity (G peak and D peak) is directly proportional to the diameter of the MWCNT, and inversely proportional to the length of the MWCNT; G peak frequency shift is closely related to the MWCNT diameter and length, which are inversely proportional to the diameter (with identical results of the single-walled carbon nanotube radial breathing modes) and direct proportional to the length. While, the influences of the diameter and length on D peak frequency shift are weak, and future analysis for the reason of this kind of phenomenon is as follows. Subsequently, we investigated the relation between D peak frequency shift and MWCNT aspect ratio, the relationship between G peak frequency shift and aspect ratio is nearly linear increase. Using the same analysis method, we plotted the different graphs of G peak and D peak intensity vs the aspect ratio of MWCNT, respectively. As the expected, the linear degression relation are existent in the two relationships.