Objective:To explore the protective effect and the underlying mechanism of Hu-Lu-Ba-Wan(葫芦巴丸,HLBW)on the testis of diabetic rats.Methods:Twenty-four male Wistar rats(160–180 g)were randomly divided into 3 groups according to a random number table,including a control group(n=8),diabetic group(n=8),and HLBW group(n=8).Diabetic rat model was established by high-fat-diet administration and single intravenous injection of streptozotocin(26 mg/kg).Then HLBW granule was administrated for 12 weeks.Fasting blood glucose and insulin levels as well as serum total testosterone level and testicular testosterone content were examined.Oxidative stress markers in both serum and testis were tested.Meanwhile,testicular morphology was observed under hematoxylin and eosin(HE)and the ultrastructure of Leydig cell was observed by electron microscope.The superoxide anion level was detected by DHE,and TUNEL-positive cells of testis was evaluated by TUNEL assay.The gene and protein expression of protein kinase C(PKCα),phosphorylated PKCα(P-PKCα)and P47 phox in testicular tissues were determined by quantitative RT-PCR analysis and Western bolt analysis.Results:Compared with the diabetic group,HLBW treatment significantly reduced the fasting glucose levels and increased the levels of fasting insulin and testosterone in serum(P<0.01).HLBW administration also reduced the levels of reactive oxygen species(ROS)in plasma and alleviated the damage of oxidative stress in the testis of diabetic rats.Additionally,HLBW down-regulated the protein and m RNA levels of PKCα,P-PKCαand P47 phox in testicular tissues.Conclusion:HLBW may attenuate the oxidative stress in the testis of diabetic rats via PKCα/NAPDH oxidase signaling pathway.
Objective： To investigate the effects of berberine （BBR） and cinnamic acid （CA）, the main active components in Jiaotai Pill （交泰丸, JTP）, on palmitic acid （PA）-induced intracellular tdglyceride （TG） accumulation in NIT-1 pancreatic 13 cells. Methods： Cells were incubated in culture medium containing PA （0.25 mmol/L） for 24 h. Then treatments with BBR （10 μmol/L）, CA （100 μmol/L） and the combination of BBR and CA （BBR＋CA） were performed respectively. Intracellular lipid accumulation was assessed by Oil Red O staining and TG content was measured by colorimetric assay. The expression of adenosine monophosphate-activated protein kinase （AMPK） protein and its downstream lipogenic and fatty acid oxidation genes, including fatty acid synthase （FAS）, acetyl-coA carboxylase （ACC）, phosphorylation acetyl-coA carboxylase （pACC）, carnitine acyl transferase 1 （CPT-1） and sterol regulating element binding protein lc （SREBP-lc） were determined by Western blot or real time polymerase chain reaction. Results： PA induced an obvious lipid accumulation and a significant increase in intracellular TG content in NIT-1 cells. PA also induced a remarkable decrease in AMPK protein expression and its downstream targets such as pACC and CPT-I. Meanwhile, AMPK downstream lipogenic genes including SREBP-lc mRNA, FAS and ACC protein expressions were increased. Treatments with BBR and BBR＋CA, superior to CA, significantly reversed the above genes changes in NIT-1 pancreatic 13 cells. However, the synergistic effect of BBR and CA on intracellular TG content was not observed in the present study. Conclusion： It can be concluded that in vitro, BBR and BBR＋CA could inhibit PA-induced lipid accumulation by decreasing lipoqenesis and increasin.cl lipid oxidation in NIT-1 pancreatic B cells.
The effect of Fructus Mume formula and its separated prescription extract on insulin resis- tance in type 2 diabetic rats was investigated. The rat model of type 2 diabetes was established by feed- ing on a high-fat diet for 8 weeks and by subsequently intravenous injection of small doses of strepto- zotocin. Rats in treatment groups, including the Fructus Mume formula treatment group （FM）, the cold property herbs of Fructus Mume formula treatment group （CFM）, the warm property herbs of Fructus Mume formula treatment group （WFM）, were administrated with Fructus Mume formula and its sepa- rated prescription extract by gavage, while the rats in diabetic model group （DM） and metformin group （MET） were given by gavage with normal saline and metformin correspondingly. The body weight be- fore and after treatment was measured, and the oral glucose tolerance test （OGTT） and the insulin re- lease test （IRT） were performed. The homeostasis model assessment-insulin resistance index （HOMA-IR） was calculated. The protein and mRNA expression levels of Insr, β-arrestin-2, Its-1 and Glut-4 in the liver, skeletal muscle and fat tissues were detected by using Western blotting and RT-PCR respectively. The results demonstrated that, as compared with DM group, OGTT, IRT （0 h, 1 h） levels and HOMR-IR in treatment groups were all reduced, meanwhile their protein and mRNA expression levels of Insr, Irs-1 and Glut-4 in the liver, skeletal muscle and fat tissues were obviously increased, and their protein and mRNA expression levels of β-arrestin-2 in the liver and skeletal muscle tissues were also markedly increased. It was suggested that the Fructus Mume formula and its separated prescription extracts could effectively improve insulin resistance in type 2 diabetic rats, which might be related to the up-regulated expression of Insr, Irs-1 and Glut-4 in the liver, skeletal muscle and fat tissues, and β-arrestin-2 in the liver and skeletal muscle tissues.
