全杜仲胶囊在犬股骨头坏死修复过程中对VEGF/bFGF的调控作用和对Bcl-2/Bax及Caspase-3蛋白表达的影响

目的:探究全杜仲胶囊对犬股骨头坏死的修复治疗作用。方法:将12只比格犬随机分成正常组、模型组、全杜仲胶囊组、仙灵骨葆胶囊组,每组3只。除正常组其余各组均通过液氮交替冷冻建立股骨头坏死模型。正常组与模型组在造模期间不予任何干预,全杜仲组在造模当天即开始进行全杜仲灌胃;仙灵骨葆胶囊组在造模当天即进行仙灵骨葆胶囊灌胃,每日1次,连续12周。比较各组第12周血管VEGF、bFGF的水平表达量,并采用Micro CT测量BMD、BS/BV、BV/TV、Tb.Th、Tb.N的比值,以及用免疫反应检测出Bcl-2、Bax、Caspase-3蛋白的表达量。结果:(1)与正常组比较,模型组造模第12周后血清VEGF、bFGF水平降低(P<0.05);与模型组比较,仙灵骨葆胶囊组、全杜仲胶囊组造模第12周血清VEGF、bFGF水平均升高,结果具有统计学差异(P<0.05),全杜仲胶囊组水平最高,其次是仙灵骨葆胶囊组。(2)与正常组比较,模型组BMD、BS/BV、BV/TV、Tb.Th、Tb.N均将低,Tb.Sp升高,结果均具有统计学意义(P<0.05)。与模型组比较,仙灵骨葆胶囊组,全杜仲胶囊组BMD、BV/TV、Tb.Th、Tb.N均升高,BS/BV、Tb.Sp降低(P<0.05)。(3)全杜仲胶囊组和仙灵骨葆胶囊组能显著提高狗股骨头中bcl-2蛋白的表达,与模型组比较有明显差异(P<0.05)。全杜仲胶囊组和仙灵骨葆胶囊组能明显降低狗股骨头中bax蛋白的表达,与模型组比较有显著差异(P<0.05)。全杜仲胶囊组和仙灵骨葆胶囊组能明显降低狗股骨头中caspase-3蛋白的表达,与模型组有显著差异(P<0.05)。结论:全杜仲胶囊可通过升高血清中VEGF、bFGF表达含量,提高bcl-2的表达量,抑制bax的表达量,并且降低caspase-3的表达量,对治疗股骨头缺血性坏死起着协同的作用,存在潜在的作用靶点。

中医治疗膝骨关节炎研究进展

膝骨关节炎(KOA)是软骨下骨发生退行性改变的一类关节疾病,中医属“痹证”“骨痹”范畴,轻者疼痛、活动受限,重者会导致关节畸形,甚至不能行走,是临床骨科疾病中发病数量较多的一类疾病。多数医家认为该病属“本虚标实”,肝肾亏虚为本、风寒湿邪为标,且与肝、脾、肾等脏腑密切相关。临床中对于膝骨关节炎的治疗方式有很多,方药、针灸、推拿、外敷等均是被证明有效的治疗方式。而随着中医水平的不断提高发展,中医的各项治疗手段注定成为临床中不可或缺的一部分。笔者从膝骨关节炎病因病机、中医药物治疗、针灸练功等方面总结膝骨关节炎研究进展,对膝骨关节炎的治疗与辨证进行总结与概括。

Regulation of Quan Du Zhong capsule on VEGF/bFGF and expression of Bcl‑2/Bax and Caspase‑3 protein in the repairing process of canine femoral head necrosis

Objective:To explore the repair and treatment effect of Quan Du Zhong capsule on necrosis of femoral head in dogs.Methods:Totally 12 beagles were randomly divided into normal group,model group,Quan Du Zhong capsule group and Xianlinggubao capsule group,with three in each group.In addition to the normal group,the other groups established the femoral head necrosis model by liquid nitrogen alternative freezing.The normal group and the model group did not have any intervention during the modeling period,and the Quan Du Zhong group began to receive the Quan Du Zhong by gavage on the day of modeling;Xianlinggubao capsule group was given Xianlinggubao capsule by gavage once a day for 12 consecutive weeks on the day of modeling.The levels of VEGF and bFGF in the blood vessels of each group at the 12th week were compared,and the ratios of BMD,BS/BV,BV/TV,Tb.Th,Tb.N were measured by Micro CT,and the expressions of Bcl-2,Bax,Caspase-3 proteins were detected by immune reaction.Results:1.Compared with the normal group,the level of serum VEGF and bFGF in the model group decreased after 12 weeks of modeling(P<0.05);Compared with the model group,the levels of serum VEGF and bFGF water in the Xianlinggubao capsule group and the Quan Du Zhong capsule group increased on average at the 12th week of modeling,with statistical difference(P<0.05).The level of the Quan Du Zhong capsule group was the highest,followed by the Xianlinggubao capsule group.2.Compared with the normal group,BMD,BS/BV,BV/TV,Tb.Th and Tb.N in the model group were lower,and Tb.SP were higher,the results were statistically significant(P<0.05).Compared with the model group,the BMD,BV/TV,Tb.Th and Tb.N of Xianlinggubao capsule group and the total eucommia capsule group increased,while the BS/BV and Tb.SP decreased(P<0.05).3.The Quan Du Zhong capsule group and Xianlinggubao capsule group could significantly increase the expression of bcl-2 protein in the femoral head of dogs,which was significantly different from the model group(P<0.05).The expression of bax protein in the femoral head of dogs in the Quan Du Zhong capsule group and the Xianlinggubao capsule group was significantly reduced compared with the model group(P<0.05).The expression of caspase-3 protein in the femoral head of dogs was significantly reduced in the Quan Du Zhong capsule group and Xianlinggubao capsule group,which was significantly different from the model group(P<0.05).Conclusion:Quan Du Zhong capsule can increase the expression of VEGF and bFGF in serum,increase the expression of bcl-2,inhibit the expression of bax,and reduce the expression of caspase-3,which plays a synergistic role in the treatment of avascular necrosis of the femoral head,and has potential targets.

药用昆虫喙尾琵琶甲肠道抗菌活性放线菌的筛选及鉴定

【目的】探究药用昆虫喙尾琵琶甲Blaps rynchopetera肠道具有抗菌活性的放线菌,为抗菌药物开发提供新的放线菌资源。【方法】结合稀释涂布法和选择培养法从喙尾琵琶甲成虫肠道分离放线菌。以耐甲氧西林金黄色葡萄球菌(methieillin-resistant Staphylococus aureus)、金黄色葡萄球菌、大肠杆菌Escherichia coli、粪肠球菌Enterococcus faecalis、鼠伤寒沙门氏菌Salmonella typhimurium和铜绿假单胞菌Pseudomonas aeruginosa 6株致病细菌以及黑曲霉Aspergillus niger、青霉菌Penicillium expansum和白色念珠菌Canidia albicans 3株致病真菌为指示菌,通过牛津杯琼脂扩散法测试放线菌次生代谢产物的抗菌活性。随后,采用分子生物学方法进行16S rRNA序列分析鉴定活性显著的18株放线菌并构建系统发育树。【结果】从喙尾琵琶甲成虫肠道中分离到176株共生放线菌,初步活性筛选结果显示其中46株放线菌表现出不同程度的抗菌活性。多株放线菌对致病菌具有广谱的抑菌作用且抑菌圈直径超过阳性对照药。选择抑菌圈直径大于15 mm的18株放线菌进行分子鉴定,结果显示均为链霉菌属Streptomyces。【结论】喙尾琵琶甲肠道含有丰富的抗菌活性放线菌资源。

美洲大蠊成虫肠道抗细菌共生放线菌的筛选及鉴定

【目的】鉴于野外美洲大蠊Periplaneta americana对恶劣环境适应性强,本研究旨在从云南省大理州的野外美洲大蠊成虫肠道中分离、筛选出抗细菌活性放线菌,为抗生素开发提供菌种资源。【方法】采用涂布平板法和平板划线法对美洲大蠊成虫肠道放线菌进行分离;以金黄色葡萄球菌Staphylococcus aureus、耐甲氧西林金黄色葡萄球菌、铜绿假单胞菌Pseudomonas aeruginosa、粪肠球菌Enterococcus faecalis、大肠杆菌Escherichia coli和鼠伤寒沙门氏菌Salmonella typhimurium共6种人体病原细菌为指示菌株,采用牛津杯法对分离自这些放线菌的次生代谢产物进行抗菌活性测定;通过形态学特征和16S rRNA基因序列分析,对具有广谱和明显抗菌活性的放线菌进行鉴定,并经16SrRNA基因序列的BlAST同源性比对及系统发育分析确定它们的分类地位。【结果】从美洲大蠊成虫肠道共分离获得41株放线菌。抗菌活性测定结果表明,34株(82.9%)放线菌对至少1种指示病原细菌具有抑制作用,其中有7株对3种以上病原细菌具有抑制作用,9株表现出明显的抗菌活性。根据系统发育分析结果,这些放线菌均被鉴定为链霉菌属Streptomyces spp.。【结论】野外美洲大蠊成虫肠道含有丰富的抗细菌活性的放线菌资源,为后续挖掘新型抗生素提供重要的微生物资源。