Notoginsenoside R_1 interferes with renal oxidative stress and Nrf2/HO-1 signaling pathway to alleviate kidney injury in mice with IgA nephropathy and its mechanism / 中国中药杂志

The purpose of this study was to investigate the effect of notoginsenoside R_1(NGR_1) on alleviating kidney injury by regulating renal oxidative stress and the Nrf2/HO-1 signaling pathway in mice with IgA nephropathy(IgAN) and its mechanism. The mouse model of IgAN was established using a variety of techniques, including continuous bovine serum albumin(BSA) gavage, subcutaneous injections of carbon tetrachloride(CCl_4) castor oil, and tail vein injections of lipopolysaccharide(LPS). After successful modeling, mice with IgAN were randomly separated into a model group, low, medium, and high-dose NGR_1 groups, and a losartan group, and C57BL6 mice were utilized as normal controls. The model and normal groups were given phosphate buffered saline(PBS) by gavage, the NGR_1 groups were given varying dosages of NGR_1 by gavage, and the losartan group was given losartan by gavage for 4 weeks. The 24-hour urine of mice was collected after the last administration, and serum and kidney tissues of mice were taken at the end of the animal experiment. Then urine red blood cell count(URBCC), 24-hour urine protein(24 h protein), serum creatinine(Scr), and blood urea nitrogen(BUN) levels were measured. The enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of galactose-deficient IgA1(Gd-IgA1), kidney injury molecule 1(Kim-1), and neutropil gelatinase-associated lipocalin(NGAL) in the mouse serum. The assay kits were used to detect the levels of malondialdehyde(MDA) and superoxide dismutase(SOD), and immunofluorescence(IF) was used to detect the expression level of glutathione peroxidase 4(GPX4) in the mesangial region. Western blot was used to detect the protein expression of nuclear transcription factor E2 related factor 2(Nrf2)/heme oxygenase 1(HO-1) signaling pathway in the renal tissue. Hematoxylin-eosin(HE) staining was used to observe pathological alterations in the glomerulus of mice. The results revealed that, as compared with the model group, the serum Gd-IgA1 level, URBCC, 24 h protein level, renal damage markers(Kim-1 and NGAL) in the high-dose NGR_1 group decreased obviously and renal function indicators(BUN, Scr) improved significantly. The activity of SOD activity and expression level of GPX4 increased significantly in the high-dose NGR_1 group, whereas the expression level of MDA reduced and protein expression levels of Nrf2 and HO-1 increased. Simultaneously, HE staining of the renal tissue indicated that glomerular damage was greatly decreased in the high-dose NGR_1 group. In conclusion, this study has clarified that NGR_1 may alleviate the kidney injury of mice with IgAN by activating the Nrf2/HO-1 signaling pathway, improving antioxidant capacity, and reducing the level of renal oxidative stress.

Effect of leech on lipid metabolism and liver in hyperlipidemia rats / 中国中药杂志

To explore the effect of leech on lipid metabolism and liver function in hyperlipidemia rats and the possible mechanism, biochemical analyzer was used to examine the regulation of leech on levels of serum triglycerides(TG), total cholesterol(TC), low-density lipoprotein cholesterol(LDL-C), and high-density lipoprotein cholesterol(HDL-C). The levels of ALT and AST in serum were detected by ELISA. The proteins expression of ACAT-2, Fas and HMGCR in liver tissue was detected by Western blot. The weight of body and liver were weighed, and liver index was calculated. Oil red O staining was used to observe the lipid accumulation in liver tissue of rats by light Microscope. The results showed that leech could decrease the levels of TC, LDL-C obviously, and increase HDL-C, decrease the levels of ALT, AST and the liver index, down-regulate the proteins expression of ACAT-2, Fas and HMGCR. And oil red O staining indicated that the lipid accumulation was less in the liver tissue of the rats intervented by leech. These data indicated that leech may affect the expression of ACAT-2, Fas and HMGCR in liver tissue to reduce the synthesis of cholesterol and fatty acid, and promote the cholesterol transforming, then regulate lipid metabolism to decrease the levels of serum lipid, and reduce lipid accumulation in liver tissue and ease liver injury of rats, then slowing down the process of nonalcoholic fatty liver disease(NAFLD) in hyperlipidemia rats.

Effect of combination of Penthorum chinense Pursh and Puerariae flos on L-02 liver cell injury induced by alcohol / 中国病理生理杂志

AIM:To explore the effect of Penthorum chinense Pursh and Puerariae flos-containing serum on L-02 liver cell injury induced by alcohol and its possible mechanism. METHODS:After preparing drug-containing serum, the L-02 cells cultured in vitro were divided into 6 groups:blank control group, model group, 1∶1 group, 2∶1 group and 1∶2 group of combination of Penthorum chinense Pursh and Puerariae flos, and tiopronin group. The viability of the L-02 cells was measured by MTT assay. The activity of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and superoxide dismutase (SOD), and the content of malondialdehyde ( MDA) were detected by enzyme label methods. The expression of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) at mRNA and protein levels was determined by real-time PCR and Western blot, respectively. RE-SULTS:Compared with control group, the levels of ALT, AST and MDA were increased significantly, and SOD was de-creased in model group ( P <0.01). Compared with model group, these indexes in all treatment groups were opposite (P<0.01). Compared with control group, the expression of TNF-α and IL-6 at mRNA and protein levels was significantly increased, the mRNA and protein expression of Nrf2 and HO-1 was decreased (P<0.01). Compared with model group, these indexes in combination groups were opposite (P<0.01). CONCLUSION:The therapeutic effects of Pentehorum chinensa Pursh and Puerariae flos-containing serum may affect the expression levels of TNF-α, IL-6, Nrf2 and HO-1, and reduce the inflammatory reaction and oxidative stress in alcohol-induced L-02 liver cells, which plays a role in attenuating alcoholic liver injury.

Effect of leech on VSMCs in early atherosclerosis rats via p38MAPK signaling pathway / 中国中药杂志

To explore the effect of leech on proliferation and apoptosis of vascular smooth muscle cells(VSMCs) in early atherosclerosis rats via p38MAPK signaling pathway and investigate its possible mechanism. Biochemical analyzer was used to examine the regulation of leech on levels of triglycerides(TG), total cholesterol(TC), low-density lipoprotein(LDL-C), and high-density lipoprotein(HDL-C) in blood lipid of rats. The expression of transforming growth factor-beta 1(TGF-β1) in serum was detected by ELISA. Immunological histological chemistry (IHC) was taken to measure the expression levels of proliferating cell nucleus antigen(PCNA) and cell apoptosis proteinase-3(Caspase-3), while the protein expression levels of MKK3, p38 and C-myc were detected by Western blot. In addition, hematoxylin and eosin(HE) staining was used to observe the morphological change of thoracic aortas. The results showed that leech decreased the levels of TC, LDL-C obviously and increased HDL-C, suppressed the expression levels of TGF-β1 and PCNA, up-regulated Caspase-3, down-regulated the expression levels of MKK3, p38, and C-myc protein. HE staining indicated that it could inhibit intimal thickening and reduce plaque formation. The above results indicated that leech may affect the protein expression of the p38MAPK signaling pathway to inhibit proliferation and promote the apoptosis of VSMCs via reducing blood lipid levels and suppressing TGF-β1, aiming at inhibiting intimal thickening and reducing plaque formation, tand then slowing down the process of early atherosclerosis.