| The epidemic of diabetes,obesity,hyperlipidemia and other diseases related to glucose and lipid metabolism has become the focus of general attention.In recent years,it has been found that people who often stay up too late,work shifts and jet lag were prone to diabetes,obesity,hyperlipidemia and other related diseases,indicating that there may be a close relationship between the change of circadian rhythm and glucose and lipid metabolism.However,the pathogenesis is still unclear and may be related to insulin resistance,energy metabolism imbalance,clock genes,inflammation and other factors.Chinese medicine think that if the Yin of liver and kidney could be damaged by staying up too late,the syndrome of deficiency of Qi and Yin would be appear.In order to further explore the influence of circadian rhythm changes on glucose and lipid metabolism,the experiment used light of different rhythms to simulate the changes of circadian rhythm,and added Qi Kui Granules for intervention treatment,so as to reveal the molecular mechanism of Qi Kui Granules for nourishing Qi and Yin,clearing heat and humidity.Because guinea pigs are more sensitive to light source,and guinea pigs are closer to humans in blood lipid and other aspects,guinea pigs are selected as the experimental model,which is the innovation of this experiment.Objective:The effects of different light rhythms on glucose and lipid metabolism in guinea pigs were observed.On the basis of the results of the preliminary experiment,Qi Kui Granules and Melatonin were added to the guinea pig group with continuous effects of light for 4 weeks,and the relevant indexes in the tissues and serum were analyzed after treatment.At the same time,the level of Clock Genes changes in peripheral tissues including liver,adipose and muscle,as well as changes level of AMPKα/PPARα signaling pathway were detected.The aim of this study is to reveal that the molecular mechanism of Qi Kui Granules improve glucose and lipid metabolism disorder caused by different light rhythms.Methods:1)A guinea pig model with different light effects on high-fat feeding in different rhythms was established,that is,control group,high-fat group,12H group and 24H group,to observe the changes of food intake,water consumption,body weight,FBG,OGTT,ITT,blood lipid,endocrine hormone and other relevant indicators.2)Guinea pig models with different light effects on rhythm and high-fat diet were repeatedly constructed,the guinea pigs of the 24H group were randomly divided into the 24H group,the Chinese medicine group and Melatonin group.Under the condition of original light and high-fat diet,the Chinese medicine group and the Melatonin group were treated with Qi Kui Granules or Melatonin respectively for 4 weeks.3)The changes in serum levels of MLT,INS,ADP,LEP,TNF-α,IL-6 and IL-1β were detected by Elisa.4)Histopathological methods were used to observe the morphological changes of liver and adipose tissue in guinea pigs after treatment.5)Fluorescence quantitative PCR was used to find the changes of Clock Genes in peripheral tissues of liver,adipose and muscle after treatment.6)The expressions of AMPKα,p-AMPKα,PPARα,CPT1A and other genes and proteins levels in liver and adipose tissue are detected by Western Blot and Fluorescence Quantitative PCR after treatment.Results:1)At the 6th week,food intake,water consumption,body weight,FBG,OGTT,ITT,blood lipid and endocrine hormone of 24H group were significantly higher than those in the 12H group with statistical significance(P<0.05).2)After 4 weeks of intervention,the food intake,water consumption,body weight,FBG,OGTT,ITT,blood lipid and endocrine hormone of the Chinese medicine group and the Melatonin group were significantly improved and the insulin sensitivity was significantly enhanced compared with that of the 24H group with a statistical difference(P<0.05).3)In liver and adipose tissue of HE staining and oil red staining,the liver lipoid lesions in the 24H group were more serious than those in the 12H group with increasing lipid accumulation in the liver.The morphology of adipose cells in the 24H group was significantly increased and the boundary was blurred compared with that in the 12H group,but the pathological changes of liver and adipose cells in the 24H group were significantly improved after treatment.4)Under the influence of continuous light,Clock Genes in liver,adipose and muscle were significantly inhibited in the 24H group compared with the 12H group.After treatment of Chinese medicine or Melatonin,the Clock Genes in liver,adipose tissue and muscle were up-regulated as a whole compared with that in the 24H group.The expressions of CLOCK,BMAL1 and RORa were most significant increase,and PER2 was the most significant decrease(P<0.05).5)Western blot and fluorescence quantitative PCR results in liver and adipose tissue of each group were compared.It was found that the expressions of AMPKα,p-AMPKα,PPARα,CPT1A gene and protein in the liver of the 24H group were significantly down-regulated compared with that of the 12H group.However,after treatment with Chinese medicine and Melatonin,these inhibited genes and proteins were significantly increased,with statistical difference(P<0.05).The expressions of adipose tissue AMPKα,p-AMPKα,PPARα,CPT1A,GLUT4 gene and protein in the 24H group were significantly decreased compared with that in the 12H group,while the expressions of these genes and protein in the Chinese medicine and Melatonin group were significantly improved,showing statistical differences(P<0.05).Conclusion:1)Prolonged light duration could aggravate insulin resistance and accelerate the formation of glucose and lipid metabolism disorders.2)Qi Kui Granules could regulate the glucose and lipid metabolism disorders of guinea pigs.It indicated that Qi Kui Granules may have a good therapeutic effect on the disorder of glucose and lipid metabolism.3)Prolonged light exposure could disrupt the levels of melatonin,adiponectin and leptin.After treatment with Qi Kui Granules,these hormone levels tended to be normal,indicating that Qi Kui Granules had a certain effect on improving the hormone levels of melatonin,adiponectin and leptin in serum.4)Qi Kui Granules had certain anti-inflammatory effect,reducing the level of TNF-α,IL-1βand IL-6 in serum.5)Qi Kui Granules could improve the expression of Clock Genes in liver,adipose tissue and muscle,which may be related to the reduction of insulin resistance.6)Prolonged light duration could result in inhibition of AMPKα/PPARα pathway in liver and fat.After treatment with Qi Kui Granules,the AMPKα/PPARα signaling pathway in liver and adipose tissue was activated,indicating that the effect of Qi Kui Granules on improving insulin resistance could be related to the activation of AMPKα/PPARα signal pathway. |
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