Protective Effect of Tangshen Formula () on Interstitial Cells of Cajal in Colon of Diabetic Rats / 中国结合医学杂志

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.

Tangshen formula promotes cellular cholesterol efflux in HG-induced mTECs by suppression of TGF-β1/Smad3 pathway / 中国临床药理学与治疗学

AIM: To observe the effect of Tangshen formula (TSF) treatment on TGF-β1/Smad3 pathway and cellular cholesterol efflux and explore its potential mechanism in HG-induced mouse tubular epithelial cells (mTECs). METHODS: After 25 mmol/L high glucose induced mTECs, TSF and Smad3 inhibitor SIS3 were given to intervene respectively. The lipid content in the cells was detected by ELISA kit; TGF-β1/Smad3 pathway and PGC-1α, LXR, ABCA1, ABCG1 were detected by Western blot and real-time PCR. RESULTS: TSF diminished the levels of TC, TG, LDL-C and increased the levels of HDL-C in HG-induced mTECs. Western blot and real-time PCR analysis showed that expression levels of TGF-β1, Smad3, Collagen and Fibronectin were significantly downregulated in the HG-induced mTECs with TSF and SIS3 treatment. And PGC-1α, LXR, ABCA1, ABCG1 expression levels were significantly upregulated in the HG-induced mTECs with TSF and SIS3 treatment. CONCLUSION: TSF can promote the cellular cholesterol efflux in HG-induced mTECs vis suppression of TGF-β1/Smad3 pathway.

Tangshen formula improves cholesterol uptake and efflux of macrophages induced by high lipid via activating PGC-1α/LXR/ABCA1 pathway / 中国临床药理学与治疗学

AIM: To observe the effects of Tangshen formula (TSF) treatment on lipid efflux and uptake in sodium palmitate (PA) induced RAW264.7 macrophages. METHODS: After 200 μmol/L PA induced RAW264.7 macrophages, TSF and PGC-1α-siRNA were given to intervene respectively. The lipid content in the cells was detected by ELISA kit; intracellular lipid droplet deposition was detected by BODIPY 493/503 and Filipin staining. Western blot and Real-time PCR were used to detect the expression of PGC-1α, LXR, ABCA1 and CD36. RESULTS: TSF diminished the levels of TC, TG and intracellular lipid droplet deposition in PA-induced RAW264.7 macrophages. Western blot and Real-time PCR analysis showed that TSF could up-regulate the expression of PGC-1α, LXR, ABCA1 and down-regulate the expression of CD36. Furthermore, silencing PCG-1α by SiRNA significantly suppressed the effects of upregulating the expression of PGC-1α, LXR and ABCA1, and downregulating the CD36 expression with TSF treatment. CONCLUSION: TSF may extenuate intracellular lipid droplet deposition in macrophages by upregulating cholesterol efflux through activating the PGC-1α/LXR/ABCA1 pathway and inhibiting lipid uptake through down-regulateing the expression of CD36.

Experimental study of Tangshen formulain improved lipid metabolism and phenotypic switch of macrophage in db/db mice / 中国中药杂志

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.

Effects of Tangshen Formula on liver oxidative stress and pathologic changes in typeⅡdiabetic rats / 天津医药

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.