The Effect Of Doucha Cassia Granules On Inflammatory Factors And Inflammatory Pathways Induced By Obesity

Chapter One Effects of Cassia nomame granules on inflammatory factors and inflammatory pathway in obese C57BL/6J mice induced by high fat diet Objective:To study the effect of Cassia nomame granules on the body weight and lipid,the related inflammatory factors and inflammatory pathway in obese C57BL/6J mice induced by high fat diet and to explore its mechanism.Methods:1.After 2 weeks of adaptive feeding,90 C57BL/6J mice were given high fat diet for 2weeks except for the control group,then randomly divided into 5 groups: model group,Cassia nomame granules group(LD?MD?HD;0.1,0.3,0.9g/kg),rosiglitazone group(RH).2.Except for the control group,the other 5 groups were given high-fat diet and weighed weekly.The hair color,state,diet and urine stool of the mice were observed daily.After 8weeks,blood was taken from eyeball and fasting blood glucose,insulin and blood lipid were measured.The fat of abdomen,epididymis and kidney of mice were taken out,the surface fluid was absorbed by filter paper,weighed by electronic balance,and refrigerated at-80 ?for reserve.3.Serum total cholesterol(TC),triglyceride(TG),high density lipoprotein(HDL-C)and low density lipoprotein(LDL-C)levels were measured by biochemical kit.4.The contents of insulin and related inflammatory factors were detected by ELISA method.5.The expression of IKK-?/NF?B and JNK pathway proteins were detected by protein immunoblotting method(Western Blot).Results:1.Through the daily observation of mice,it was found that the hair of each group of mice was bright,active,exploratory,good appetite,normal feces,no diarrhea and so on.2.Compared with the control group,the weight of the model group was significantly higher than that of the control group,and there was significant difference at the end of the 8th week(p < 0.01).Compared with the model group,the inhibitory effect of HD and RH group was the most obvious(p < 0.01),and there was no significant change between LD group and model group(p < 0.05).The results showed that Cassia nomame granules could reduce the weight gain in mice induced by high-fat.3.Compared with the control group,the fasting blood glucose(FPG),insulin(INS)and insulin resistance index(IRI)in the model group were significantly higher than those in the control group.Compared with the model group,FPG,INS and IRI were significantly decreased in LD,MD,HD group and RH group.The results showed that Cassia nomame granules could significantly reduce FPG,INS and IRI in mice induced by high-fat.4.Compared with the control group,low density lipoprotein(LDL-C)and total cholesterol(TC),)triglyceride(TG)in the model group were significantly higher,high density lipoprotein(HDL-C)decreased significantly(P < 0 01)than those in the control group(P < 0 01).Compared with the model group,MD,HD and RH groups decreased LDL-C,TC,TG and elevated the level of HDL-C.Compared with the model group,LD group decreased the level of LDL-C and increased the level of HDL-C.Cassia nomame granules has a regulating effect on blood lipid,with the increase of dosage,the more obvious the effect.5.Compared with the control group,tumor necrosis factor ?(TNF-?),interleukin-6(IL-6)and serum leptin(leptin,LEP),monocyte chemoattractant protein-1(MCP-1)were significantly increased(p < 0.01),the levels of adiponectin(ADPN)were significantly decreased in the mice fed with high fat diet.Compared with the model group,the TNF-?,IL-6 LEP,MCP-1 were significantly decreased(p < 0.01),ADPN level increased significantly in mouse serum of LD,MD,HD,RH group.6.The results of Western Blot showed that the ratio of p-IKK ?/IKK ? and p-JNK/JNK,the level of NF ? B were increased in the model group fed with high fat diet.However,the levels of p-IKK ?/IKK ?,NF ? B and pJNK in the administration group were significantly lower than those in the model group.The results showed that Cassia nomame granules could inhibit the expression of IKK-?/NF?B and JNK pathway protein and inhibit the activation of IKK-?/NF?B and JNK pathway induced by obesity.Conclusion:1.Cassia nomame granules feed mice skin hair bright,active,energetic,normal feces,no diarrhoea,anorexia,and so on.Cassia nomame granules can significantly reduce the weight of high-fat-fed mice.2.Cassia nomame granules can improve glucose and lipid metabolism and insulin resistance in obese C57BL/6J mice induced by high fat diet.3.Cassia nomame granules can inhibit the secretion of related inflammatory factors and the activation of inflammatory pathway in obese C57BL/6J mice induced by high fat,and improve insulin resistance.Chapter two Effects of containing serum of Cassia nomame granules on inflammatory factors and inflammatory pathway of 3T3-L1 preadipocytes Objective:To observe the effect of Cassia nomame granules containing serum on inflammatory factors and inflammatory pathway of 3T3-L1 preadipocytes.Methods:1.3T3-L1 preadipocytes were cultured with 10% fetal bovine serum(FBS),0.1% penicillin and streptomycin high glucose DMEM medium using cell culture technique.0.5mmol/L3-isobutyl-1-methylxanthine(IBMX),1 ? mol/L dexamethasone(DEX)and 10 ? g/mL insulin(Ins)were used to induce 3T3-L1 cell differentiation.Oil red O staining was used to detect the results of differentiation of 3T3-L1 cell.2.The differentiated preadipocytes of 3T3-L1 were divided into blank control group,Cassia nomame granules containing serum group(5%,10%,20%)and cultured for 24 hours,collected after 24 hours of LPS(LPS)stimulation.The levels of inflammatory factors TNF-a,IL-6,IL-1? and MCP-1 were measured by Elisa method.3.The differentiated 3T3-L1 preadipocytes were divided into groups and stimulated by LPS for 24 h.The total protein content of the cells was measured by BCA method.The expression of IKK-?/NF?B and JNK pathway protein were detected by Western Blot.Results:1.The differentiation of 3T3-L1 preadipocytes was induced by cocktail method.Oil red O staining was used to detect the differentiation results.The results showed that more than 85%of the cells showed obvious "ring exchange" oil droplets,and 3T3-L1 preadipocytes were induced to differentiate into mature adipocytes.2.The results of Elisa showed that the secretion of TNF-a,IL-6,IL-1?and MCP-1 increased significantly in 3T3-L1 cells treated with LPS.The secretion of TNF-a,IL-6,IL-1? andMCP-1 in 3T3-L1 cells cultured with Cassia nomame granules containing serum was significantly decreased.3.The results of Western Blot showed that the expression of pIKK-?/ IKK-?,NF?B and p-JNK/JNK protein was significantly increased in the mature 3T3-L1 cells treated with LPS.The expression of pIKK-?/ IKK-?,NF?B and pJNK/JNK in 3T3-L1 cells were cultured with the serum containing of Cassia nomame granules was significantly decreased.Conclusion:1.3T3-L1 preadipocytes were induced into the mature adipocytes by cocktail method for 8 ~12 days.2.The serum containing of Cassia nomame granules significantly decreased the secretion of inflammatory factors and inhibited the activation of IKK-?/NF ? B and JNK pathway in differentiated 3T3-L1 cells stimulated by LPS.