Tinospora crispa extract inhibits MMP-13 and migration of head and neck squamous cell carcinoma cell lines

Objective: To investigate the effect of Tinospora crispa(T. crispa) extract on matrix metalloproteinase 13(MMP-13) expression and cell migration. Methods: The cytotoxicity of T. crispa extract was examined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay on head and neck squamous cell carcinoma(HNSCC) cell lines. The effect on expression of MMP-13 was analysed by RT-PCR and ELISA. The migration was assessed by wound healing assay. Results: MMP-13 m RNA was highly expressed in the metastatic human HNSCC cell lines, HN22 and HSC-3. T. crispa extract at a concentration of 100.0 μg/m L caused about 50% reduction of cell survival. T. crispa extract at a non-toxic concentration of 12.5, 25.0 and 50.0 μg/m L signii cantly suppressed MMP-13 m RNA expression and secreted MMP-13 in both HN22 and HSC-3. The expression of tissue inhibitors of metalloprotease by HSC-3 cells was attenuated by 25.0 and 50.0 μg/m L of T. crispa extract. Addition of the extract to cells in a wound healing assay showed inhibition of cell migration by HN22 cells. Conclusions: These data suggest that T. crispa could be considered as a potential therapeutic drug to prevent metastasis of HNSCC.

cis-Clerodane Diterpenoids with Structural Diversity and Anti-inflammatory Activity from Tinospora crispa

Twelve new cis-clerodane diterpenoids, designated as crispinoids A—L (1—12), together with three known analogues (13—15), were isolated from Tinospora crispa. Their structures were fully assigned by comprehensive spectroscopic techniques, single-crystal X-ray diffraction experiments, and electronic circular dichroism (ECD) analyses. The isolated clerodanes displayed diverse heterocyclic frameworks including 6/6/6-, 6/5/6/6-, 6/6/5-, 6/6-, and 6/5/6-fused ring systems. Some of the isolates showed ATP-citrate lyase (ACLY) and nuclear factor kappa B (NF-κB) pathway inhibition. The NF-κB inhibitors further suppressed the lipopolysaccharide (LPS)-induced inflammatory responses in RAW 264.7 cells.

波叶青牛胆提取物对糖尿病肾病小鼠药效及急性毒性

目的研究波叶青牛胆的乙醇提取物和不同极性溶剂萃取部位对Ⅱ型糖尿病肾病小鼠的疗效以及急性毒性。方法 100只采用高脂饮食联合小剂量链脲佐菌素(STZ)复制Ⅱ型糖尿病肾病成功的ICR小鼠分为模型组,乙醇提取物低、中、高剂量组,三氯甲烷部位组,乙酸乙酯部位组,正丁醇部位组,水溶性部位组,以及卡托普利组和消渴丸阳性药组,并设正常对照组。给药6周末测定各组小鼠血清中总胆固醇、尿素氮、肌酐、胱抑素C水平,PAS染色观察肾脏病理变化。结果乙醇提取物的LD50为4 887 mg/kg。与模型组相比,乙醇提取物高剂量组和正丁醇部位组能显著改善糖尿病肾病小鼠各血清生化指标水平,缓解肾小球肥大,肾小球基底膜增厚及系膜扩张等病理改变。结论波叶青牛胆属于低毒类中草药,在其安全剂量范围内可以降低糖尿病肾病小鼠的血糖、总胆固醇,改善肾功能状态。

波叶青牛胆胰蛋白酶抑制剂的纯化及其性质研究

采用亲和层析及电泳制备等方法 ,从防已科植物波叶青牛胆 (Tinosporacrispa)根茎中分离到一种胰蛋白酶抑制剂TCTI。对其性质研究表明 :TCTI的相对分子量约为 10 0kD ,对牛胰蛋白酶的抑制常数Ki为 2 0 8× 10 7mol/L (BAPNA为底物时 ) ,摩尔抑制比为 1∶3 5 ,是一种抑制活力很强的竞争性抑制剂。TCTI具有很高的耐热性 ,在 10 0℃加热 60min ,仍保持 93%的抑制活力。另外 。

不同处理对波叶青牛胆扦插的影响

以波叶青牛胆的茎段为插条,采用三因素三水平正交实验设计,研究基质、ABT浓度以及浸泡时间对波叶青牛胆扦插成活的影响,并在正交试验最优组合的基础上做进一步优化试验。结果表明,3种因素对波叶青牛胆生根的影响从大到小依次为A(基质)、C(浸泡时间)、B(ABT浓度),优组合为A_(1)B_(2)C_(1);对发芽效果的影响从大到小为C(浸泡时间)、A(基质)、B(ABT浓度),发芽效果的优组合为A_(2)B_(3)C_(1);对扦插效果的影响从大到小为A(基质)、C(浸泡时间)、B(ABT浓度),优组合为A_(1)B_(2)C_(1);优化实验中,最优组合为河沙+ABT 400 mg/L+浸泡2~10 min,此处理组合的波叶青牛胆生根率高、腐烂率低、根系发达、粗壮、发芽率较高,且植株健壮。