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Objective To establish a mouse model of kidney-yin and kidney?yang deficiency after oral administra?tion of hydrocortisone, and to explore the related evaluation factors. Methods The model was established by oral adminis?tration of hydrocortisone to induce kidney?yin and kidney?yang deficiency in mice. The survival and body weight of the mice were observed. The serum content of adrenal cortical hormone ( ACTH) , cortisol ( Cor. ) in the hypothalamic?pituitary?ad?renal (HPA) axis, and thyroid stimulating hormone (TSH), triiodothyronine (T3), thyrox (T4) in the hypothalamic?pi?tuitary?thyroid (HPT) axis, follicle?stimulating hormone (FSH), estradiol (E2), testosterone (T) in the hypothalamic?pituitary?gonadal ( HPG) axis were determined by radioimmunoassay. Results The body weight of kidney?yin and kidney?yang mice were decreased, the serum ACTH, Cor, TSH, T3, T4 contents were decreased, the serum FSH, E2, T contents were increased in the kidney?yang deficiency model mce ( P<0. 01 ) , and those parameters in the kidney?yin deficiency model mice were changed in opposite direction. Conclusions It is found that the hormone levels of ACTH, Cor, TSH, T3, T4, FSH, E2 and T in kidney deficiency mice are changed, and cortisol can be used as an important index to evaluate the model of kidney deficiency induced by glucocorticoid.
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Objective To observe the effects ofLiuwei Dihuang Decoction on cAMP and PDE3B in adipose tissues of rats with type 2 diabetes;To explore its mechanism.MethodsTotally 80 SD rats were randomly divided into two groups:control group and molding group. Rats in the molding group established the type 2 diabetic models by feeding high sugar and fat diet combined with intraperitoneal injection of Streptozotocin. The successful modeling rats were divided into model group, Rosiglitazone group, andLiuwei Dihuang Decoction group. Administration groups were given relevant medicine for gavage, while model group and blank group were given the same amount of normal saline for 30 days. FBG levels were detected, and cAMP and INS levels were detected by ELISA. The protein expressions of PDE3B in adipose tissues were determined by Western blot. The mRNA expressions of PDE3B in adipose tissues were detected by RT-PCR.Results Compared with the model group, the FBG and INS levels in type 2 diabetic rats were reduced in Rosiglitazone group andLiuwei DihuangDecoction group (P<0.05,P<0.01). InLiuwei Dihuang Decoction group, the protein and mRNA expressions of PDE3B in the adipose tissues were higher, and cAMP levels were lower than the model group (P<0.01).ConclusionLiuwei Dihuang Decoction can increase PDE3B expression, and decrease cAMP level, which may be one of the mechanisms for improving insulin resistance in type 2 diabetic rats.
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Objective To construct a rat model of nonalcoholic fatty liver disease (NAFLD) by feeding rats fat-rich diet and analyze the effect of insulin resistance (IR)in the development of NAFLD. Methods Male SD rats were randomly divided into normal diet group (NG, n =24) and fat-rich diet group (FG, n =24). At the end of feeding for2 weeks, 4 weeks, 6 weeks or 8weeks, 6 rats in NG and FG were randomly took out. Their weight were recorded, then the serum fasting blood sugar, fasting insulin, triglyceride, total cholesterol, ala-nine aminotransferase and aspartate aminotransferase were measured, and the fasting insulin resistance index and liver index (liver weight (g)/body weight(g) × 100%)was calculated. Then liver tissues were homogenized, and muleic dialdehyde and superoxide dismutase were determined. The hepatic steatosis in all rats was assessed according to the results under light microscope. Results The body weight of rats in NG increased faster than those in FG after six weeks. The liver index of rats in FG was markedly higher than that in NG since the second weekend. The rats in FG began to have hepatocyte steatosis from the second weekend, had insulin resistance, hyperlipidemia, dysfunction of liver and lipid peroxide of liver from the fourth weekend, suffered mild fatty liver from the sixth weekend, and developed to moderate fatty liv-er from the eighth weekend. Conclusions NAFLD with IR model was successfully developed by feeding SD rats with an improved rich-fat diet for 6 weeks. IR may play an important role in the development of NAFLD.
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Objective To construct and identify replication deficient recombinant adenovirus expressing human c-Jun N-terminal kinase(JNK)by homologous recombination adenovirus dominant-negative type JNK(Ad-DN-JNK).Methods The linearized recombinant shuttle vector pAdTrack-CMV-DN-JNK Was co-transformed with backbone vector pAdEasy-l into bacteria BJ5183 for recombinant adenoviral vector.The recombinant adenoviral vector was transfected into HEK293 packing cells tO construct replication deficient recombinant adenovirus,and then the recombinant edenovirns WaS detected by PCR and DNA sequencing.Western blot analysis was utilized to detect the Cxpression of Ad-DN-JNK and the level of insulin receptor substrate l Serine307 phosphorylation.Results JNK recombinant adenoviral vectorcould be effectively transfeeted into HEK 293 cell and successfully packed by intracellular enzyme.The expression of green fluorescent protein(GFP)Was observed on the 5th day after transfection.The fragment of JNK gene waS amplified by PCR and identified by sequencing.The titer of the prepared Ad-DN-JNK is 2.5×1010 pfu/ml.The animal experiment confirmed that constructed Ad-DN-JNK could be effectively expressed in liver tissue.Conclusion The research successfully constructed recombinant adenoviral vector and recombinant adenoviral particle.Animal experiment demonstrated the Ad-DN-JNK could effectively mediated the expression of DN-JNK gene and down-regulated the level of IRSlscfine307 phosphorylation.The achievement laid a foundation for further investigation of the function and application of JNK.
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Objective To construct replication deficient recombinant adenovirus expressing human c-Jun N-terminal kinase by homologous recombination.Methods The linearized recombinant shuttle vector pAdTrack-CMV-WT-JNK was co-transformed with backbone vector pAdEasy-1 into bacteria BJ5183 for recombinant adenoviral vector.The recombinant adenoviral vector was transfected into HEK293 packing cells to construct replication deficient recombinant adenovirus,and then the recombinant adenovirus was detected by PCR and DNA sequencing.Results JNK recombinant adenoviral vector was effectively transfected into HEK 293 cells and was successfully packed by intracellular enzyme.The expression of green fluorescent protein(GFP)was observed on the 5th day after transfection.The fragment of JNK gene was amplified by PCR and identified by sequencing.The animal experiment confirmed that Ad-WT-JNK was effectivety expressed in liver tissue. Conclusion The research successfully constructed recombinant adenoviral vector and recombinant adenoviral particle.And the achievement laid a foundation for further investigation of the function and application of JNK.