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ObjectiveTo explore the underlying mechanism of Tripterygium wilfordii polyglycoside tablets （TWPT） in the prevention and treatment of kidney injury in diabetic nephropathy （DN） through the nuclear factor of activated T-cells 2（NFAT2）/cyclooxygenase-2（COX-2） pathway. MethodForty-two male SD rats of SPF grade were selected and randomly divided into a normal group （n=8） and an experimental group （n=34） after one week of adaptive feeding. The rats in the normal group were fed conventionally. The DN model was established in rats of the experimental group by intraperitoneal injection of streptozotocin （STZ） following one week of feeding on a high-fat and high-glucose diet. After the death and failure cases during modeling were eliminated， the remaining 24 model rats were randomly divided into model group， valsartan （8.33 mg·kg-1·d-1） group， and TWPT （5 mg·kg-1·d-1） group. Rats in normal group and model group were given equal amounts of normal saline by gavage. After six weeks， body weight was measured and urine samples were collected. Blood samples were collected from the abdominal aorta， and then the rats were sacrificed for sampling. Biochemical indicators， such as serum blood urea nitrogen （BUN）， serum creatinine （SCr）， alanine aminotransferase （ALT）， blood lipid， blood glucose， and 24-hour urine total protein （24 h UTP）， were determined. Hematoxylin-eosin （HE） staining and Masson staining were used to observe the pathology of the kidney. Enzyme-linked immunosorbent assay （ELISA） was used to detect NFAT2 and COX-2 expression levels in the serum. Western blot and Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）were adopted to detect NFAT2， COX-2 protein and mRNA expression in kidney tissues， respectively. ResultCompared with the normal group， the model group showed elevated 24 h UTP， BUN， SCr， CHO， TG， and FBG， increased serum NFAT2 and COX-2 production and expression （P<0.01）， and elevated protein and mRNA expression of NFAT2 and COX-2 in kidney tissues （P<0.01）. In addition， the pathology of the kidney showed enlarged glomeruli， mild proliferation of mesangial cells， and widened mesangial stroma. Compared with the model group， the TWPT group showed decreased 24 h UTP， BUN， SCr， CHO， TG， and FBG （P<0.05，P<0.01）， basically normal glomerular morphology， decreased expression of serum NFAT2 and COX-2 （P<0.01）， and down-regulated protein and mRNA expression of NFAT2 and COX-2 in kidney tissues （P<0.01）. ConclusionTWPT can alleviate 24 h UTP in DN model rats， protect renal function， and improve renal pathology， and its mechanism of action may be related to the down-regulation of NFAT2/COX-2 expression in the serum and kidney tissues.
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BACKGROUND:Previous studies have found that combined use of Buyang Huanwu decoction and bone marrow mesenchymal stem cel (BMSC) transplantation can play a synergic role against cerebral ischemia injury. OBJECTIVE:To analyze the effect of Buyang Huanwu decoction combined with BMSC transplantation to promote angiogenesis after cerebral ischemia. METHODS:Ninety-six Sprague-Dawley rats were randomly divided into four groups, and used to make middle cerebral artery occlusion models. In combined group, rats were given intragastrical administration of Buyang Huanwu decoction 10 mL/kg once a day, beginning at 3 days prior to modeling, and then given intragastrical administration of Buyang Huanwu decoction once at 2 hours after modeling, fol owed by its intragastrical administration every 12 hours. In BMSC and combined groups, BMSC suspension was injected into the rat ventricle after 2-hour cerebral ischemia/2-hour reperfusion, and then 30 minutes later, CD34 and CD45 antibodies were injected. In antibody group, CD34 and CD45 antibodies were injected. In model group, only normal saline was given. SerumαVβ3 level detection, immunohistochemical observation, Q-PCR and western blot tests were performed in the combined group at 12, 24, 36, 48 hours after reperfusion, while these indices were detected in the other three groups at 36 hours after reperfusion. RESULTS AND CONCLUSION:(1) The level of serumαVβ3 was lower in the antibody group than the model group (P<0.05), higher in the BMSC and combined groups than the antibody group (P<0.05), and higher in the combined group than the BMSC group (P<0.05). (2) Immunohistochemical findings showed that compared with the antibody group, the number of CD34 positive cel s was higher in the model, BMSC and combined groups (P<0.05). (3) Results from the Q-PCR and western blot assay showed that compared with the model group, the pFAK protein expression level was lower in the antibody and BMSC group, but FAK gene expression level had no overt changes;while the protein levels of FAK (24 hours after reperfusion) and pFAK (12 hours after reperfusion) were significantly increased in the combined group than the antibody and BMSC groups (P<0.05, P<0.01). Moreover, this increase exhibited a gradual y rising trend with the extension of reperfusion time. To conclude, the combined use of BMSC transplantation and Buyang Huanwu decoction can reverse the effect of CD34+CD45 antibodies that lead to the decrease in the number of vascular endothelial cel s and levels of integrinαVβ3 and downstream signaling molecules, thereby to promote angiogenesis in the MCAO model.
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Objective Mitochondrial DNA depletion syndrome is a rare autosomal recessive disorder characterized by complex genetic and clinical manifestations.This study aimed to investigate the clinical and laboratory features of a boy with mitochondrial encephalomyopathy caused by SUCLG1 mutation.Methods The clinical data and genetic test of a patient with mitochondrial DNA depletion syndrome were retrospectively analyzed.Result The proband presented with limb weakness at the 4th month after birth,and presented dystrophic appearance,muscular hypotonia,psychomotor retardation,failure to thrive,hearing impairment,scoliosis,thoracocyllosis and facial features at 9 months old.Laboratory tests showed blood lactic acid and pymvate increased,liver damage and abnormal myocardial enzymes.Plasma camitine ester profiling showed that amino acids decreased and C4-dicarboxylic-carnitine increased.Urinary organic acid analysis showed increased concentration of methylmalonic acid and its metabolites indicating methyl malonic aciduria.MRI showed bilateral T2 hyperintensities in bilateral caudate nuelei and lenticular and brain atrophy-like changes.Brainstem auditory evoked potential showed severe hearing loss.His development quotient was 35.Genetic sequencing of MUT,,MMAA,MMAB and other classic mitochondrial disease related genes of the proband revealed no mutation.Two heterozygous mutations,c.961C＞G and c.713T＞C,inherited from the phenotype of normal parents were detected in his SUCLG1 gene.The copy number of mitochondrial DNA was 244/cell in peripheral blood leukocytes,equivalent to 68.4％ of that in normal control.Conclusion In this study,an infant with muscular hypotonia,psychomotor retardation,deafness and slightly increased urine methyl malonic acid was diagnosed by genetic test.For patients with unexplained hypotonia,mental retardation,abnormal movements,hearing disorder together with increased blood pyruvic acid and lactic acid,mild methylmalonic acidemia and abnormal acylcarnitine,mitochondrial DNA depletion syndrome should be considered.Gene analysis is important for diagnosis and prenatal diagnosis of the next pregnancy.