朱顶红无菌苗叶片高效再生体系

以朱顶红（Hippeastrum vittatum）叶片为外植体进行离体培养，具有取材方便、试材充足、成本低等优势，但叶片诱导再生率极低，是朱顶红离体培养的一大难题。本试验中分别以‘花孔雀’和‘黑天鹅’朱顶红无菌苗叶片为外植体，探究了不同植物生长调节剂和不同取材部位对不定芽诱导和继代增殖的影响。结果表明：最佳外植体为 MS 培养基中培养 10 d 形成的幼嫩叶片基部（0.5 cm），在光照 16 h · d-1（光照强度 36 μmol · m-2 · s-1）下，不定芽诱导的最适培养基为 MS + 2 mg · L-1 6-BA + 1 mg · L-1 NAA + 2 mg · L-1 TDZ，两个品种的不定芽均以间接途径发生，其中‘花孔雀’在培养 40 d 后形成愈伤组织，55 d形成不定芽，诱导率可达 69.44%；‘黑天鹅’在培养 45 d 后形成愈伤组织，65 d 形成不定芽，诱导率达到 66.67%；最适体细胞胚诱导培养基为 MS + 2 mg · L-1 6-BA + 1 mg · L-1 PIC，‘花孔雀’和‘黑天鹅’的诱导率分别达到 66.67%和 63.89%；最佳不定芽增殖培养基为 MS + 2 mg · L-1 6-BA + 1 mg · L-1 NAA + 1 mg · L-1 TDZ，‘花孔雀’和‘黑天鹅’的增殖系数分别达到 4.67 和 3.46；在不添加植物生长调节剂的 MS培养基中进行生根培养，30 d 后两个品种的生根率均达到 100%；将生根培养 30 d 的小植株转移至室温条件下放置 3 d，摘去封口膜再驯化 3 d 后，移栽至经高温消毒的草炭︰蛭石（体积比）为 1︰1 的基质中，成活率达到 100%

Role of SGK1 inhibitor EMD638683 in preventing renal tissue pyroptosis induced by bladder outlet obstruction in mice

AIM: To study the mechanism of serum and glucocorticoid-regulated kinase 1 (SGK1) inhibitor EMD638683 against inflammation of obstructive nephropathy, and to explore the relationship between SGK1 and NLRP3-caspase1-IL-1β/pyroptosis pathway-induced cell damage in a mouse model of lower urinary tract obstruction-induced bladder outlet obstruction (BOO). METHODS: After the BOO model was constructed, the inflammatory cell infiltration was observed by HE staining, the tissue lesions of the kidney were observed by PAS staining, the collagen deposition of renal tubules was detected by Masson staining, and the expression levels of NLRP3, caspase-1, F4/80, gasdermin D N-terminal fragment (GSDMD-N), IL-1β and SGK1 were determined by immunohistochemistry and Western blot. The mouse renal tubular epithelial cells (mRTECs) were used, and treated with aldosterone (ALD) and EMD638683. The expression levels of NLRP3, caspase-1, IL-1β, SGK1 and GSDMD-N were detected by Western blot. The IL-1β content in the cell supernatant was determined by ELISA. RESULTS: After obstruction for 3 weeks in the mice, the infiltration of inflammatory cells was observed in the kidney by HE staining, renal tissue lesions were observed by PAS staining, and collagen deposition was detected in tubulointerstitium by Masson staining. Compared with normal group, the levels of NLRP3, caspase-1, F4/80, GSDMD-N, IL-1β and SGK1 were significantly increased in the 3-week obstructed mice (P<0.05 or P<0.01). The expression levels of NLRP3, caspase-1, IL-1β, SGK1 and GSDMD-N in mRTECs were increased significantly in ALD group (P<0.05 or P<0.01). Compared with ALD group, the expression of NLRP3, caspase-1, IL-1β, SGK1 and GSDMD-N were significantly reduced in ALD+ EMD638683 group (P<0.05 or P<0.01). CONCLUSION: We successfully constructed renal inflammatory injury mediated by BOO-induced lower urinary tract obstruction in mice. EMD638683 blocked pyroptosis and exerted its anti-inflammatory effect by inhibiting NLRP3-caspase-1-pyroptosis pathway.