ABSTRACT
OBJECTIVE@#To explore the effect of Tangshen Formula (, TSF), a Chinese herbal medicine, on interstitial cells of Cajal (ICC) in the colon of diabetic rats.@*METHODS@#Fifty-four male Wistar rats were randomly divided into normal control (NC, n=14) and high-fat diet (HFD) groups (n=40). After 6 weeks, the rats in the HFD group were injected intraperitoneally streptozotocin once (30 mg/kg). Thirty rats with fasting blood glucose higher than 11.7 mmol/L were randomly divided into diabetes (DM) and TSF groups, 15 rats in each group. Rats in the NC and DM groups were intragastrically administered with saline, and those in the TSF group were given with TSF (2.4 g/kg) once daily for 20 weeks. Expression levels of Bax, Bcl-2, and caspase-3 in colonic smooth muscle layer were measured by Western blotting and immunohistochemical staining. The number of ICC was determined by immunohistochemical staining. Immunofluorescence was used for analyzing the ratio of classically activated macrophages (M1) and alternatively activated macrophages (M2) to total macrophages. Electron microscopy was used to observe the epithelial ultrastructure and junctions.@*RESULTS@#TSF appeared to partially prevented loss of ICC in DM rats (P<0.05). Compared with the NC group, expression levels of Bcl-2, Bax, caspase-3, and TNF-α as well as the ratio of M1 to total macrophages increased in DM rats (all P<0.05), and the ratio of M2 to total macrophages decreased (P<0.05 or P<0.01). Compared with the DM group, TSF decreased the expression levels of abovementioned proteins and restore M2 to total macrophages ratio (P<0.05 or P<0.01). TSF appeared to attenuate the ultrastructural changes of epithelia and improve the tight and desmosome junctions between epithelia reduced in the DM rats.@*CONCLUSION@#Reduced number of ICC in DM rats may be associated with damage of the intestinal barrier. The protective effects of TSF on ICC may be through repair of the epithelial junctions, which attenuates inflammation and inflammation-initiated apoptosis in colon of DM rats.
Subject(s)
Animals , Male , Rats , Colon , Diabetes Mellitus, Experimental/drug therapy , Drugs, Chinese Herbal/therapeutic use , Interstitial Cells of Cajal , Rats, WistarABSTRACT
Obesity and its associated metabolic disorders, including non-alcoholic fatty liver disease (NAFLD), have become major chronic diseases threatening public health. NAFLD is a chronic liver disorder that is strongly associated with type 2 diabetes and obesity. In this study, we investigated the effects and mechanism of Tangshen formula (TSF) on hepatic dyslipidemia and phenotypic switch of macrophage in db/db mice. Eight-week-old male C57BLKS/J db/m control and db/db mice were divided into 3 groups (namely db/m, db/db, db/db+TSF), and fed with TSF or distilled water for 12 weeks. It was found that after treatment with TSF, the triglycerides accumulation in db/db mice was decreased on the basis of oil red O staining with cryosections of liver tissues. And protein expressions of macrophage activation markers CD68 and F4/80 were decreased according to immunohistochemical analysis of hepatic sections. The mRNA level of TNF-α (M1 marker) was significantly decreased by TSF in db/db mice, but with no significant difference in Mrc1 and Arg1 (M2 marker). According to the results, TSF attenuated hepatic steatosis and relieved dyslipidemia, its mechanism may be correlated with the regulation of macrophage activation and phenotypic switch.
ABSTRACT
Objective To investigate the effects of Tangshen Formula on liver oxidative stress in diabetic rats, and their mechanisms thereof.Methods Rat model of type 2 diabetes mellitus and nonalcoholic fatty liver disease was estab?lished by intraperitoneally injecting small dose of chain urea with cephalosporins (STZ) and feeding high fat fodder . The model rats were randomly divided into control group, model group, Tangshen Formula group and metformin group.The se?rum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride (TG), cholesterol (CHO), low density lipoprotein-cholesterol(LDL-C), and superoxide dismutase (SOD), catalase(CAT), vitamin E(VE), nitric oxide (NO) and nitric oxide synthase (NOS) in liver were compared between four groups. Changes of pathological morphology were ob?served under light microscopy. Results There were significantly decrease in serum levels of ALT, AST, TG, CHO, LDL-C in Tangshen Formula group and metformin group compared with those of model group. There were significantly higher levels of SOD, CAT and VE, and lower levels of NO and NOS of liver homogenate in Tangshen Formula group than those of model group. There were higher levels of SOD and CAT, and lower levels of NO and NOS in liver homogenate in metformin group than those of model group. HE staining showed that liver fatty degeneration was significantly reduced in metformin group and Tangshen Formula group compared with that of model group. Conclusion The fatty liver in type 2 diabetic rats is signifi?cantly improved by Tangshen Formula treatment, which is probably associated with the regulation of the response level to oxi?dative stress in liver.