| Objective: The clinical effect of Yun-Fu-Jiang-Zhuo(YFJZ)massage combined with Xiao-Xian-Xiong Decoction(XXXD)on the glucose and lipid metabolism disorder in patients with obese type 2 diabetes mellitus(T2DM)(phlegm heat syndrome)was observed,then a T2 DM rat model was established to further verify the effect and analyze the difference of YFJZ massage,XXXD and their combination in improving the glucose and lipid metabolism disorder,and the underlying mechanism was explored based on oxidative stress and inflammatory response,so as to provide a basis for its application in the prevention and treatment of T2 DM.Methods:1.Clinical study on the effect of YFJZ massage combined with XXXD on the glucose and lipid metabolism disorder in obese T2DM(phlegm heat syndrome)Outpatients in Affiliated Hospital of Changchun University of Traditional Chinese Medicine from April 2020 to April 2021 were randomly divided into metformin group,metformin + massage group(combined massage group)and metformin + massage + XXXD group(comprehensive treatment group),36 cases in each group.Patients in the three groups were asked to receive the health education on diabetes,control their diet,take regular exercise and undergo the other basic treatment,in which patients in the metformin group were treated with Metformin Hydrochloride Sustained-Release Tablets orally,those in the combined massage group with YFJZ massage and those in the comprehensive treatment group with YFJZ massage combined with XXXD.After treatment,the changes of relevant indexes in the three group were compared,mainly including routine indexes,biochemical indexes,TCM symptom score,curative effect and adverse reactions.2.Experimental study on the effect of YFJZ massage combined with XXXD on the glucose and lipid metabolism disorder in T2 DM ratsA T2 DM rat model was established by giving rats a high-fat diet combined with the intraperitoneal injection of STZ.The rats were randomly divided into model group,metformin group,massage group,XXXD group and combined treatment group,and another normal group was set,10 rats in each group.After the intervention treatments,the general state of all rats was observed,their body weights were measured,the content of FBG and the level of FINS were detected,and the HOMA-IR values were calculated.At the same time,the serum TG,TC,LDL-C and HDL-C levels were detected,and the liver histopathological changes were examined by HE staining.3.Effects of YFJZ massage combined with XXXD on the oxidative stress and inflammatory response in T2 DM ratsSOD,GSH-Px and CAT activities,and MDA content in the liver tissue of rats were measured by xanthine oxidase,dithio-dinitrobenzoic acid,ammonium molybdate and thiobarbituric acid methods,respectively.TNF-α,IL-1β and IL-6 contents in the liver tissue were measured by ELISA.NADPH oxidase(NOX2,NOX4),AMPK/Nrf2/HO-1 signaling pathway proteins(p-AMPKα,AMPKα,Nrf2 and HO-1),IKK/NF-κB signaling pathway proteins(p-IKKβ,IKKβ,IκBα and NF-κBp65)and IRS-1/PI3K/GLUT4 signaling pathway proteins [p-IRS-1(Tyr),p-IRS-1(Ser),IRS-1,PI3Kp85 and GLUT4] in the liver tissue were detected by Western blot.Results:1.Clinical study on the effect of YFJZ massage combined with XXXD on the glucose and lipid metabolism disorder in obese T2DM(phlegm heat syndrome)(1)Routine indexes: Compared with those before treatment,some routine indexes of patients in the different groups,such as the waist circumference,body weight and BMI,decreased significantly after treatment(P < 0.05).After treatment,compared with that in the metformin group,the waist circumference of patients in the combined massage group and comprehensive treatment group decreased significantly(P < 0.05),while there was no significant difference in the body weight and BMI although they decreased to a certain extent(P > 0.05);Compared with those in the combined massage group,there was no significant difference in the waist circumference,body weight and BMI in the comprehensive treatment group(P > 0.05).The above results indicate that the treatment with YFJZ massage alone or YFJZ massage combined with XXXD may help to reduce the abdominal fat,but can not significantly affect the body weight and BMI in obese T2 DM patients with phlegm heat syndrome.(2)Biochemical indexes:(1)Glucose metabolism indexes: Compared with those before treatment,the contents of FBG,2h PG and Hb A1 c decreased significantly in the different groups after treatment(P < 0.05);After treatment,compared with those in the metformin group,the contents of FBG,2h PG and Hb A1 c in the combined massage and comprehensive treatment groups decreased significantly(P < 0.05);Compared with those in the combined massage group,the contents of FBG,2h PG and Hb A1 c in the comprehensive treatment group were further reduced(P < 0.05).(2)Lipid metabolism indexes: Compared with those before treatment,the contents of TG,TC and LDL-C in the different groups decreased significantly after treatment(P < 0.05),while the content of HDL-C increased significantly(P < 0.05);After treatment,the contents of TG,TC and LDL-C in the combined massage and comprehensive treatment groups were significantly lower than those metformin group(P < 0.05),and the content of HDL-C was not significantly different from that in the metformin group(P > 0.05);Compared with those in the combined massage group,the contents of TG,TC and LDL-C in the comprehensive treatment group were further reduced(P < 0.05),and there was no significant difference in the content of HDL-C between the two groups(P > 0.05).(3)Inflammation indexes: Compared with those before treatment,the contents of TNF-α,IL-6 and CRP in the different groups decreased significantly after treatment(P < 0.05);After treatment,compared with those in the metformin group,the contents of TNF-α,IL-6 and CRP decreased significantly in the combined massage and comprehensive treatment groups(P < 0.05);Compared with those the combined massage group,the contents of TNF-α,IL-6 and CRP in the comprehensive treatment group decreased further(P < 0.05).The above results suggest that YFZJ massage combined with XXXD may have a more significant effect to alleviate the glucose and lipid metabolism disorder and inhibit the inflammatory response in obese T2 DM patients with phlegm heat syndrome.(3)TCM symptom scoring and curative effect: Compared with that before treatment,the TCM symptom score in the different groups decreased significantly after treatment(P <0.05).After treatment,compared with those in the metformin group,TCM symptom scores in the combined massage and comprehensive treatment groups decreased significantly(P <0.05);Compared with that in the combined massage group,the TCM symptom score in the comprehensive treatment group was further reduced(P < 0.05);The total effective rates in the three groups were 69.44%,80.56% and 88.89%,respectively,and no severe adverse reactions were found in all the patients.2.Experimental study on the effect of YFJZ massage combined with XXXD on the glucose and lipid metabolism disorder in T2 DM rats(1)Glucose metabolism: Compared with that in the model group,the content of FBG in the massage,XXXD and combined treatment groups decreased significantly after treatment(P < 0.05);Compared with that in the massage group,the content of FBG in XXXD and combined treatment groups decreased significantly(P < 0.05);Compared with that in XXXD group,the content of FBG in the combined treatment group was further reduced(P < 0.05);At the same time,the glucose tolerance of rats in each group was significantly improved,in which the improvement effect in XXXD group was stronger than that in YFJZ massage group,and the improvement effect in the combined treatment group was most significant.(2)Insulin secretion and insulin resistance: Compared with those in the model group,the level of FINS and value of HOMA-IR in the other groups decreased significantly(P <0.05),except that there was no significant difference in the level of FINS in the massage group(P > 0.05);Compared with those in the massage group,the level of FINS and value of HOMA-IR in XXXD and the combined treatment group were further reduced(P < 0.05);There was no significant difference in the level of FINS between XXXD group and the combined treatment group(P > 0.05),but the value of HOMA-IR decreased significantly(P< 0.05),suggesting that YFJZ massage,XXXD and the combined treatment all can improve the insulin resistance and then affect the glucose and lipid metabolism in T2 DM rats.(3)Lipid metabolism: YFJZ massage,XXXD and the combined treatment could reduce the contents of serum TG,TC and LDL-C(P < 0.05),with a good regulatory effect on the lipid metabolism disorder in T2 DM rats,but no significant effect on the content of HDL-C(P > 0.05);There is no significant difference in improving the lipid metabolism disorder in T2 DM rats between the effect of YFJZ massage and XXXD,but the effect of the combined treatment is better than that of YFJZ massage or XXXD alone.(4)Liver histopathological changes: The liver histopathological changes of rats in each group after treatment were alleviated to varying degrees compared with the model group,especially in the combined treatment group.3.Effects of YFJZ massage combined with XXXD on the oxidative stress and inflammatory response in T2 DM rats(1)Anti-oxidative stress: YFJZ massage could increase the activity of SOD,the expressions of Nrf2 and HO-1 protein in the liver tissue(P < 0.05),and reduce the content of MDA and the expression of NOX4 protein(P < 0.05),and the other indexes showed no obvious change(P > 0.05);Compared with those in the model group,the activities of SOD,GSH-Px,CAT and the expressions of p-AMPKα,Nrf2 and HO-1 protein of the liver tissue in XXXD group and the combined treatment group increased significantly(P < 0.05),and the content of MDA and the expressions of NOX2 and NOX4 protein decreased significantly(P < 0.05).Compared with that in the massage group,the activities of SOD,GSH-Px and CAT,and the expressions of Nrf2 and HO-1 protein of the liver tissue in XXXD group and the combined treatment group increased significantly(P < 0.05),while the content of MDA and the expressions of NOX2 and NOX4 protein decreased significantly(P < 0.05),and the expression of p-AMPKα protein increased only in the combined treatment group(P < 0.05).Compared with that in XXXD group,the activities of SOD,GSH-Px and CAT and the expressions of p-AMPKα and HO-1 protein of the liver tissue in the combined treatment group increased further(P < 0.05),the expression of Nrf2 protein was not significantly different(P > 0.05),and the content of MDA and the expressions of NOX2 and NOX4 protein decreased further(P < 0.05).The above results indicate that XXXD and the combined treatment have a stronger antioxidant stress ability than the massage group,and the antioxidant stress effect of the combined treatment is stronger than that of XXXD,and activating the AMPK/Nrf2/HO-1 signaling pathway may be the potential mechanism though which YFJZ massage,XXXD and the combined treatment can exert their antioxidant stress to improve the glucose and lipid metabolism disorder in T2 DM rats.(2)Inhibiting inflammatory reaction: YFJZ massage,XXXD and the combined treatment could reduce the contents of TNF-α,IL-1β and IL-6,and the expressions of p-IKKβ,p-IκBα and NF-κBp65 protein in the liver tissue of rats(P < 0.05).Compared with that in the massage group,the expression of p-IKKβ protein only in XXXD decreased(P <0.05),and there was no significant difference in the other indexes(P > 0.05).Compared with those in the massage group or XXXD group,the contents of TNF-α,IL-1β and IL-6,and the expressions of p-IKKβ,p-IκBα and NF-κBp65 protein decreased further in the combined treatment group(P < 0.05).The above results suggest that there is no significant difference in anti-inflammatory effect between YFJZ massage and XXXD,and the anti-inflammatory effect of the combined treatment is significantly stronger than that of YFJZ massage or XXXD alone,and inhibiting the activation of the IKK/NF-κB signaling pathway may be an important reason for YFJZ massage,XXXD and the combined treatment to play an anti-inflammatory role and improve the glucose and lipid metabolism disorder in T2 DM rats.(3)Effects on the insulin signaling pathway: The expressions of p-IRS-1(Tyr),PI3Kp85 and GLUT4 protein in the liver tissue increased(P < 0.05)and the expression of p-IRS-1(Ser)protein decreased(P < 0.05)in YFJZ massage,XXXD and combined treatment groups,indicating that treatments may activate the insulin signaling pathway IRS-1/PI3K/GLUT4.The further analysis on the ability of YFJZ massage,XXXD and the combined treatment to activate the insulin signaling pathway IRS-1/PI3K/GLUT4 showed that the effect of the combined treatment was the strongest,followed by that of XXXD,and that of YFJZ was slightly weaker.Results:(1)YFJZ massage combined with XXXD is safe and effective in the clinical treatment of obese T2DM(phlegm heat syndrome),effectively alleviating the glucose and lipid metabolism disorder and inhibit the inflammatory response.(2)The alleviating effect of YFJZ massage on the glucose metabolism disorder is weaker than that of XXXD,the effect of both methods on the lipid metabolism disorder is equivalent,and the combined effect of the two methods on the glucose and lipid metabolism disorder is better than that of them alone.(3)YFJZ massage,XXXD and the combined treatment can play an antioxidant stress role by activating the AMPK/Nrf2/HO-1 signaling pathway,and an anti-inflammatory role by inhibiting the IKK/NF-κB signaling pathway.The effect of alleviating the glucose and lipid metabolism disorder of YFJZ massage,XXXD and the combined treatment in T2 DM rats is related to their antioxidant stress,inhibiting inflammatory response and activating the insulin signaling pathway IRS-1/PI3K/GLUT4. |
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