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Objective To explore the role of transforming growth factor-β1/serum and glucocorticoid induced kinase 1 (TGF-β1/SGK1) signaling pathway in diabetic nephropathy (DN).Methods Totally 60 clean grade healthy male Sprague-Dawley (SD) rats were divided into normal control (Con group,n =20) and diabetic model group (diabetic group,n =40).Type 2 diabetic animal model was induced by intraperitoneal injection of streptozotocin (STZ,45 mg/kg) to SD rats.Half of rats in each group were sacrificed after implemented for 4 weeks and 8 weeks,respectively.Fasting glucose,serum urea and creatinine,as well as urine protein in 24 h were detected.Pathological changes of rats' kidney cortex tissue were examined by hematoxylin-eosin staining.The expression of SGK1 was detected by immunohistochemistry in renal tissue.The mRNA level of SGK1 was analyzed by real-time polymerase chain reaction (PCR).The protein expression of TGF-β1 and SGK1 were determined by Western blotting.Results Compared to the control group,the blood glucose of the diabetic group rats was significantly increased,kidney hypertrophy index,serum urea and creatinine,24 h urine volume and urinary microalbumin as well as kidney index were significantly higher (P ＜0.05).The mRNA expressed of SGK1,protein expression of TGF-β1 and SGK1 were also increased obviously in the renal cortex of diabetic rats (all P ＜ 0.05 vs Control).Conclusions TGF-β1/SGK1 signaling pathway is closely related to the TIF damage of kidney cortex in diabetic rats.
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Objective To explore the impact of puerarin treatment on autophagy in rats with traumatic brain injury (TBⅠ) and the underlying mechanism.Methods Seventy five Sprague-Dawley (SD) rats were randomized into 5 groups:sham group (S group,n =15),traumatic brain injury group (TBⅠ group,n =15),TBⅠ + puerarin treatment group (TBⅠ + Pue group,n =15),TBⅠ + JNK inhibitor group (TBⅠ + SP group,n =15),and TBⅠ + JNK activator + Pue (TBⅠ + An + Pue group,n =15).Feeney method was applied to make rats with TBⅠ model.Mter that,head water content and neurological deficit score (NDS) were measured and recorded at day 1,3 and 7 in each group.Western blot was used to measure the JNK activity and autophagic marker proteins,including LC3B and Beclin1.Results Compared to S group,the head water content and NDS were decreased significantly among the others (P ＜ 0.05).The head water content and NDS in TBⅠ + Pue and TBⅠ + SP groups was decreased remarkably compared with TBⅠ group.Combined with puerarin and animycin treatments failed to reduce head water content and NDS compared to the TBⅠ + Pue group.Activated autophagy could be observed in TBⅠ group compared to S group.Compared to group S,LC3Ⅱ,Beclin1 and P-JNK1 were increased significantly.Pue and SP could reduce their expressions,respectively.Combined with puerarin and animycin treatments failed to reduce LC3Ⅱ,Beclin1 and P-JNK1 compared to TBⅠ + Pue group.Conclusions Puerarin could protect rats with TBⅠ via inhibiting autophagy,JNK signal pathway could involve the process of puerarin regulating autophagy.
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Objective To observe the effect of intrathecal injection (IT) of oxycodone hydrochloride on neuropathic pain and spinal cord level of microglial c-Jun N-terminal kinase/chemokine (C-X-C motif) ligand 1 (c-JNK/CXCL) 1signal in rat model of chronic constriction injury (CCI).Methods Male Sprague-Dawley rats were randomly divided into five groups (n =40 per group):sham group (intrathecal normal saline,IT NS),CCI group (CCI + IT NS),oxy group (CCI + IT 5 μg/30 μl oxy),mino group (CCI + IT 5 μg/30 μl Minocycline),and c-JNK inhibitor group (SP group,CCI + IT 5 μg/30 μl SP600125).The lumbar intrathecal catheters were implanted in L5-6 of rats and CCI models were established as previously described.The thermal and mechanical nociceptive thresholds were assessed by paw withdrawal latency (PWL) to radiant heat and von Frey filaments.The oxycodone,minocycline and SP600125 were administered intrathecally for 3 days before surgery.The spinal cord expression of Ⅰ ba-1,p-c-JNK and CXCL1 proteins assessed by Western blot.Immunofluorescence staining was performed to examine microglia morphology and the number of Ⅰ ba-1 positives cells in dorsal horn of injured spinal cord at 7 days post-IR.Results Compared to sham group,rats in CCI group had significantly lower mechanical and thermal pain thresholds,but higher spinal proteins expression of Ⅰ ba-1,and p-c-JNK and CXCL1 (P ＜0.05).Rats in oxy group,mino group and SP group had significantly higher mechanical and thermal pain thresholds and significantly lower proteins expression of Ⅰ ba-1,p-c-JNK and CXCL1 compared to those in CCI group (at any observed time-point after ligation,but most significantly at 7days,P ＜ 0.05).At the 7days after surgery,microglial cells in CCI group transformed from the ramified shape to amoeboid macrophage-like shape by immunofluorescence staining with the increases of Ⅰ ba-1 positive cells;while the other three groups exhibited hypertrophic morphology with less number Ⅰ ba-1 positive cells (P ＜ 0.05).There were no significant differences between these three groups at any observed time (P ＞ 0.05).Conclusions Intrathecal injection of oxycodone hydrochloride can relieve CCI-induced neuropathic pain by down-regulation microglial c-JNK/CXCL1 signal in spinal cords.Provide new therapeutic targets for clinical treatment of neuropathic pain.
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Objective To investigate the expressions of phosphatidylinositol 3-kinase (PI3K), Akt and E-cadherin and their clinical significance in thyroid papillary carcinomas.Methods Expressions of PI3K, Akt, and E-cadherin were detected in 62 cases of thyroid papillary carcinomas,30 cases of thyroid goiter and 30 cases of normal thyroid by immunohistochemistry (EnVison),and simultaneously compared with age, sex, tumor size, clinical tumor node metastasis( TNM) stages, and lymph node metastasis in thy-roid papillary carcinomas.Results The expression rate of PI3K, Akt, and E-cadherin was 74.2%(46/62), 66.1%(41/62), 16.1%(10/62),respectively.Expressions of three proteins in thyroid papillary carcinomas were significantly different from those in thyroid goiter and normal thyroid tissues ( P <0.05). The lower positive rates of PI3K and Akt proteins were obtained in the group of stageⅠ~Ⅱthan that in the group of stageⅢ~Ⅳ(χ2 =4.976, P =0.026;χ2 =6.233, P =0.013).Higher positive rates of PI3K and Akt proteins were obtained in the group of lymph-node metastasis than that in group of non-lymph-node metastasis (χ2 =6.675, P =0.010;χ2 =7.511, P =0.006).Higher positive rate of E-cadherin protein was obtained in the group of stage Ⅰ~Ⅱ than that in the group of stage Ⅲ ~Ⅳ (χ2 =6.558, P =0.010 ) .Higher positive rate of E-cadherin proteins was obtained in the group of non-lymph-node metastasis than that in the group of lymph node metastasis(χ2 =5.678, P =0.017).There was significant positive correlation between expressions of PI3K and Akt through Spearman correlation analysis ( r =0.423, P <0.05).PI3K was negatively correlated with E-cadherin with Spearman correlation analysis ( r =-0.527, P <0.05).Akt was also negatively correlated with E-cadherin ( r =-0.417, P <0.05).Conclusions PI3K/Akt pathway might regulate thyroid papillary carcinoma cells proliferation, invasion and metastasis.
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Objective To investigate the effect of fluvastatin on the expressions of caspase-12,CCAAT/enhancer-binding protein homologous protein(CHOP), and c-Jun N-terminal kinases (JNK) in ischemia-reperfusion brain injury in rats.Methods Forty two rats were randomly divided into sham operation group (6 rats), ischemia-reperfusion (I/R) group (18 rats), and fluvastatin (Flu) group (18 rats).The rats of I/R and Flu groups were molded by modified Longa intraluminal thread, then put to death at 2 h occlusion and 24 h reperfusion point.Expressions of caspase-12, CHOP, and JNK were detected with immunohistochemistry and Western blot.Results Immunohistochemistry and Western blot showed that the expressions of caspase-12, CHOP, and JNK were increased at 24 h reperfusion.Compared to I/R group, the expressions of caspase-12 and CHOP in Flu group were decreased significantly (all P ＜0.01);and the expression of JNK had no difference between I/R and Flu groups(P ＞ 0.05).Conclusions The increased expression of caspase-12, CHOP, and JNK showed that endoplasmic reticulum stress was involved in the pathological process of ischemia-reperfusion brain injury.Fluvastatin could inhibit the expression of caspase12 and CHOP, and could delete endoplasmic reticulum stress (ERS) in ischemia-reperfusion brain injury.