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Objective:To explore the role of thioredoxin interaction protein (TXNIP)/NOD-like receptor protein 3 (NLRP3) pathway in renal interstitial fibrosis induced by renal ischemia-reperfusion injury (IRI) in mice.Methods:Adult male C57BL/6J mice aged 6 to 8 weeks and TXNIP knockout mice with the same genetic background were selected. The wild type mice were divided into the sham operation (Sham) group and renal IRI group. The TXNIP knockout mice were divided into the sham+TXNIP KO group and IRI+TXNIP KO group, with 12 mice in each group. The model of renal ischemia-reperfusion injury was established by clamping bilateral renal pedicles for 45 min and then restoring perfusion. The sham operation model was only dissociated bilateral renal arteries without other treatment. Blood creatinine, urea nitrogen, kidney injury molecule-1 (Kim-1) and neutrophil gelatinase-associated lipocalin (NGAL), blood transforming growth factor-β (TGF-β) and interleukin 6 (IL-6) were measured on the 1st, 7th and 28th days after reperfusion. The renal cortex was taken on the 1st and 28th days for Masson staining, in which the renal tubule-interstitial injury score was obtained. TGF-β and IL-6 mRNA expression were detected by qPCR, TXNIP, NLRP3, Pro-IL-1β, IL-1β and α-SMA protein expression were detected by Western blot, and MDA and SOD levels were detected by ELISA. Homogeneity test of variance was performed before the statistics of normal distribution measurement data, one-way ANOVA was used for the comparison between multiple groups, and LSD- t test was used for the comparison between the two groups. Results:On the 1st, 7th and 28th days after IRI, compared with the sham group, the Scr, BUN, Kim-1, NGAL, TGF-β and IL-6 were increased continuously in the IRI group ( P<0.05). On the 28th day after IRI, large areas of collagen fibers and inflammatory cell infiltration were observed in the renal interstitium of the IRI group. In the IRI group, the scores of renal tubular injury and renal interstitial fibrosis on the 28th day were significantly higher than those on the 1st day (all P<0.05). On the 1st, 7th and 28th days after IRI, compared with the IRI group, the levels of Scr, BUN, Kim-1, NGAL, TGF-β and IL-6 were significantly decreased in the IRI+TXNIP KO group (all P<0.05). On the 1st and 28th days after IRI, compared to the IRI group, the areas of collagen fibers and inflammatory cell infiltration in the renal interstitium of the IRI+TXNIP KO group were decreased. The renal tubule injury score [Day 1, (192.2 ± 62.4) vs. (103.2 ± 49.1); Day 28, (154.3 ± 93.6) vs. (64.3 ± 24.8), both P<0.05] and interstitial fibrosis score [Day 1, (7.3 ± 3.2) vs. (4.8 ± 1.7); Day 28, (12.8 ± 3.9) vs. (2.3 ± 0.8), both P<0.05] were all decreased. The expression of TGF-β, IL-6 mRNA, TXNIP, NLRP3, Pro-IL-1 β, IL-1 β and α-SMA protein in renal cortex were significantly decreased (both P<0.05). In renal cortex, MDA level was decreased and SOD level was increased (all P<0.05). Conclusions:TXNIP/NLRP3 pathway is involved in the development of renal interstitial inflammation and fibrosis after renal ischemia and reperfusion. Knockout or inhibition of TXNIP can inhibit the progression of acute renal injury to chronic renal disease.
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OBJECTIVE：To study the effects of chrysophanol on the activa tion of microglia and the expression of inflammatory factors in cerebral ischemia model rats. METHODS ：SD rats were randomly divided into sham operation group ， model group and chrysophanol high ，medium，low dose groups [7.88，3.94，1.97 mg/（kg·d）]，with 20 rats in each group （the number was complemented in cases of death or unsuccessful modeling during modeling process ）. Except for sham operation group ， middle cerebral artery occlusion model was established in other groups by improved thread method. After 2 hours of ischemia ， sham operation group and model group were intraperitoneally injected with 1 mL normal saline ，and each administration group was intraperitoneally injected with 1 mL corresponding drug ，once a day ，for 7 consecutive days. After last medication ，the score of neurological impairment was recorded ；cerebral infarction of rats was observed by TTC staining ，and the percentage of cerebral infarction area was calculated. The expression of Iba- 1 positive cells in ischemic penumbra of rats was observed by immunofluorescence staining. The expression of Notch- 1，TNF-α and ICAM-1 in the ischemic penumbra of rats were detected by Western blotting assay. RESULTS ：In sham operation group ，there was no infarction area in the brain tissue ，and the Iba- 1 positive cells in the ischemic penumbra were few and branched. Compared with sham operation group ，the infarction area of cerebral tissue in rats was obvious in model group ； the 052）number of Iba- 1 positive cells in ischemic penumbra were 〔ZQ2017003〕） increased significantly ，and they were in amoeba or round shape；the neurological impairment score ，the percentage of cerebral infarction area ， relative expression of Notch- 1， TNF-α and ICAM-1 protein in ischemic penumbra were increased significantly （P＜0.05）. Compared with m odel rats ，the infarction area of cerebral tissue in each dose group of chrysophanol was reduced to different extent ；the number of Iba- 1 positive cells in ischemic penumbra was decreased ；neurological impairment score ，the percentage of cerebral infarction area ，relative expression of Notch- 1，TNF-α and ICAM-1 protein were significantly decreased （P＜0.05 or P＜0.01）. CONCLUSIONS ：Chrysophanol has a certain protective effect on the brain tissue of cerebral ischemia model rats ，and can relieve the nerve injury. Its mechanism may be associated with inhibiting the activation of microglia and expression of inflammatory factors mediated by Notch pathway.
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Objective:To investigate the expressions of leptin receptor (OBR) and phosphorylated protein kinase B (p-AKT) in patients with diffuse large B-cell lymphoma (DLBCL) and its significance.Methods:Immunohistochemistry was applied to detect the expressions of OBR and p-AKT in tissues from 90 patients with DLBCL and 20 patients with reactive lymphoid hyperplasia (RLH) between 2010 and 2015 in Shandong Provincial Hospital Affiliated to Shandong First Medical University. Cell proliferation assay was used to detect the effect of leptin on the proliferation of SUDHL4 and SUDHL5 in DLBCL cell lines, and the expression of p-AKT in SUDHL4 and SUDHL5 after the cultured leptin was detected by using Western blot.Results:The high expression rate of OBR and p-AKT of DLBCL was 47.7% (43/90) and 27.7% (25/90), respectively, and low expression was found in 20 cases of RLH. There were statistically differences in the expressions of OBR and p-AKT in DLBCL and RLH ( P < 0.01, P = 0.027). The expressions of OBR and p-AKT were not correlated with age, gender, extranodal infiltrations, clinical staging, lactic dehydrogenase (LDH) level, B-symptom and international prognostic index (IPI) score (all P > 0.05). The expression of OBR was positively related with that of p-AKT in DLBCL patients ( r = 0.532, P < 0.05). Leptin could increase the proliferation of SUDHL4 and SUDHL5 cells and promote the expression of p-AKT. Conclusion:Leptin and OBR can promote the proliferation of DLBCL cells and may be involved in the occurrence and development of DLBCL by activating PI3K-AKT pathway.
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OBJECTIVE:To detect the contents of chlorogenic acid and rutoside in Saussurea involucrate,and to optimize the decoction and extraction technology of S. involucrate from different producing areas. METHODS:The contents of chlorogenic acid and rutoside in 10 batches of S. involucrate from different producing areas were determined by HPLC. L9(34)orthogonal experiment was used to optimize the water amount,decoction times and decoction time using comprehensive score of extraction transport rate of chlorogenic acid and rutoside as index. The verification test was also conducted. RESULTS:The contents of chlorogenic acid and rutoside in 10 batches of S. involucrate were 0.380%-0.546% and 0.334%-0.617%;the optimal decoction technology was as follows as the amout of crude material of S. involucrate 100 g,soaking for 20 min,decocting for 3 times,12,10 and 10 fold of water,decocting 45,30 and 30 min,respectively. The extraction transfer rates of chlorogenic acid and rutoside were 96.2%(RSD=2.66%,n=3)and 89.3%(RSD=3.31%,n=3)in verification test. CONCLUSIONS:For S. involucrate from different producing areas,the contents of effective components are different;optimized decoction and extraction technology is stable and feasible.
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OBJECTIVE:To clarify the bio-transformation form of apigenin in rats,and to speculate its possible metabolic path-way. METHODS:Rats were divided into blank group and medication group(apigenin 200 mg/kg,i.g.)with 6 rats in each group. Urine and feces samples were collected from 2 groups within 24 h after medication. After corresponding treatment,urine and feces samples were analyzed and detected by HPLC-IT-TOF-MSn under cation mode and anion mode. RESULTS:9 metabolites were iden-tified in urine sample of rats from medication group,i.e. 2,3-double bond reduction of apigenin (U1,U7,U8,U9),bonded to glucuronic acid(U2,U3,U4),bonded to sulphate(U5,U6,U7,U8,U9)and bonded to glucose(U2). 4 metabolites were iden-tified in feces sample of rats from medication group,i.e. 2,3-double bond reduction of apigenin(F3),bonded to glucuronic acid (F2)and bonded to glucose(F1). CONCLUSIONS:Apigenin mainly exists in form of prototype drug in rats. The reduction hap-pens on 2,3-double bond by the intestinal bacteria,and the product of apigenin boned to glucuronic acid or glucose can be formed when excreting in intestinal tract and rats in vivo,while the product of apigenin boned to sulphate can be formed only when excret-ing in rats in vivo.
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OBJECTIVE: To establishes an HPLC method for determination of the contents of Osthol and Berberine in Kangfuyan effervescent suppository. METHODS: HPLC was carried out on a column of Diamonsil (250 mm?4.6 mm,5 ?m) with mobile phase consisted of acetonitrile-0.2% phosphate acid (52∶48,Lauryl sulfonic acid sodium 0.1 g was contained in every 100 mL) with flow rate at 1 mL?min-1.The detection wavelength was set at 322 nm and the column temperature was set at room temperature. RESULTS: The linear range was 0.019 8～0.396 0 ?g(r=0.999 97) for Osthole versus 0.056 65～1.133 00 ?g(r=0.999 99) for Berberine. The average recoveries of Osthol and Berberine were 101.06% (RSD=1.10%) and 101.88%(RSD=0.98%), respectively. CONCLUSION: This method is simple, accurate and reproducible, and applicable for the quality control of Kangfuyan effervescent suppository.