Objective To investigate the effect of Ligustrazine on the expressions of FoXO3a, MAFbx, and MuRF1 indenervated skeletal muscle atrophy rats. Methods Fifty-four 8-week-old female Sprague Dawley rats were randomly dividedinto 3 groups: normal control group (group A, n=6), denervated control group (group B, n=24), and Ligustrazine interventiongroup (group C, n=24). After the denervated gastrocnemius models were established in the rats of groups B and C, sal ine andLigustrazine [80 mg/(kg·d)] were given every day by intraperitoneal injection, respectively. However, no treatment was donein group A. At 2, 7, 14, and 28 days after denervation, the wet weight of gastrocnemius was measured to calculate the ratio ofwet weight. The mRNA and protein expression levels of FoXO3a, MAFbx, and MuRF1 were detected by RT-PCR and Westernblot. Results The ratio of gastrocnemius wet weight decreased with time after denervation in groups B and C, showingsignificant differences when compared with that of group A (P lt; 0.05), and group C were significantly higher than that of groupB at 7, 14, and 28 days (P lt; 0.05). The mRNA and protein expressions of FoXO3a, MAFbx, and MuRF1 in groups B and Cwere significantly higher than those in group C at 7, 14, and 28 days (P lt; 0.05), and group C was significantly lower than groupB (P lt; 0.05). Conclusion Ligustrazine may postpone denervated skeletal muscle atrophy by reducing mRNA and proteinexpressions of FoXO3a, MAFbx, and MuRF1.
Objective To investigate the effect of tetramethylpyrazine (TMP) with a certain concentration added to vitrification solution on peripheral nerve allografts regeneration. Methods Forty-eight healthy clean SD male rats were selected as donors, and 96 healthy clean Wistar male rats as recipients, all rats being 3 months old and weighing 200-250 g. The sciatic nerves segments of 15 mm were removed from the donors, then randomly divided into 4 groups according to vitrificationsolution containing TMP. No TMP was used in group A as the control group; 100 mg/L, 200 mg/L and 400 mg/L TMP were used in group B, group C and group D, respectively. Then them were cryo-preserved at — 196 ℃ for 3 weeks. Nerve defect of 10 mm in length was made in the sciatic nerves of recipients. After rewarming, the allografts were transplanted to the corresponding rats. The gross appearance, the morphological and electrophysiological changes, the image analysis of axons and motor end-plate were detected at 4, 8, 12 and 16 weeks. Results All rates survived to the end of the experiment. The adhesion and edema of allografts in group A and group B were obvious 4 weeks after operation; then adhesion and edema was obvious in group A and were improved in the other groups 8 weeks after operation. Adhesion was observed in groups A and B; no adhesion was observed in groups C and D at 12 weeks. The number of regeneration nerve, the latent, the ampl itude, the nerve conduction velocity, the medullary sheath/μm2, the medullary sheath density/μm2 and the image analysis of axons and motor end-plate in groups A and B were significantly lower than those in groups C and D (P lt; 0.01); and there were no significant differences between groups C and D (P gt; 0.05). The observation of transmission electron microscope showed that medullated nerve fibers and myel in sheath of groups C and D were thicker than groups A and B, layers of groups C and D were clear. Conclusion The vitrification solution with 200 mg/L tetramethylpyrazine has protective effect on regeneration of peripheral nerve allografts．
In order to study the mechanism of the inhibitory effect of salvia miltiorrhiza (SM) and tetramethyl pyrazine (TP) on scar fibroblast, the DNA content of fibroblast and the all distribution in cellular cycle was measured by FCM. The hypertrophic scar tissue of chest was chosen for primary culture of fibroblast. Then this cultured cell was reacted with SM and TP. FCM was used to measure the DNA index and duration of cellular cycle. The results showed that: 1. SM and TP had little effect on DNA index, but when the concentration of drugs reached the threshold, they could increase the amount of fibroblasts in C2-M stage and the duration of G2-M stage was prolonged; 2. TP could also prolong the duration of S-stage; 3. SM and TP could prolong the multiplication time of fibroblasts and this effect was correlated postively with the dosage of drug. The conclusions were that the inhibitory effect of SM was the result of inhibiting the mitosis of cells and the cellular cycle be at a standstill in G2-M stage. The inhibitory effect of TP was due to the inhibition of synthesis and duplication of DNA and cellular mitosis, and the cellular cycle was also at a standstill in G2-M stage.
In order to investigate the inhibitory effect of salvia miltiorrhiza (SM) and tetramethyl pyrazine (TP) on scartricial fibroblast, the hypertrophic scar tissue of chest was chosen for culture of fibroblasts, and the influence of SM and TP on fibroblasts was observed, The effect of the drugs on the growth of fibroblasts, on DNA synthesis of fibroblasts and on mitosis index of fibroblasts were all determined quantitatively. The results showed: 1. SM and TP could inhibit significantly the growth of the fibroblasts, the inhibitory effect was irreversible when the concentration of the drugs reached 5 mg/ml and 500 micrograms/ml respectively; 2. SM and TP could inhibit the absorption of 3H-TdR and this effect was correlated positively to the dosage of the drugs and; 3. SM and TP could reduce the mitosis index of fibroblasts. It was concluded that SM and TP had definite depressive effect on growth of fibroblasts which was correlated positively with the concentration of drugs and duration of application. The inhibitory effect of the drugs on fibroblasts was mainly through inhibition of synthesis of DNA.
Objective To observe the effect of tetramethypyrazine (TMP) on the expression of hypoxia-related factors in human umbilical vein endothelial cells (HUVECs). Methods The second to fifth passage cultured HUVECs were divided into five groups: control group, CoCl2induced hypoxic group and 50, 100, 200 mu;mol/L TMP treatment groups. HUVECs in control group were not treated. HUVECs inCoCl2induced hypoxic group were treated with 150 mu;mol/LCoCl2for four hours. HUVECs in 50, 100, 200 mu;mol/L TMP treated groups were pretreated with 150 mu;mol/LCoCl2 for four hours, followed by treatment with 50, 100, 200 mu;mol/L TMP for eight hours. Real-time reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the mRNA levels of prolyl hydroxylase 2 (PHD2), hypoxia-induced factor-1alpha;(HIF-1alpha;) and vascular endothelial growth factor (VEGF). Protein levels of PHD2, HIF-1alpha;, and VEGF were detected using Western blot. Results Compared with the control group, theCoCl2 induced hypoxic group showed decreased mRNA and protein levels of PHD2 (t=3.734, 3.122;P＜0.05), while those of HIF-1alpha; and VEGF increased (HIF-1alpha; mRNA:t=4.589,P＜0.05; HIF-1alpha; protein:t=3.778,P＜0.05. VEGF mRNA:t=3.926,P＜0.05; VEGF protein:t=3.257,P＜0.05). Compared with theCoCl2 induced hypoxic group, 50, 100, 200 mu;mol/L TMP treated groups showed increased mRNA and protein levels of PHD2 (PHD2 mRNA: t=3.286, 3.617, 3.886;P＜0.05. PHD2 protein: t=2.813, 3.026, 3.078; P＜0.05); while those of VEGF decreased (VEGF mRNA: 50 mu;mol/L TMP: t=1.696,P＞0.05; 100 mu;mol/L TMP:t=2.974,P＜0.05; 200 mu;mol/L TMP: t=3.492,P＜0.05; VEGF protein: 50 mu;mol/L TMP: t=1.986,P＞0.05; 100 mu;mol/L TMP: t=2.976,P＜0.05; 200 mu;mol/L TMP:t=3.136,P＜0.05); although changes in HIF-1alpha;mRNA levels were not statistically significant (t=1.025, 0.726, -1.386;P＞0.05), showed a decrease in HIF-1alpha;protein levels (50 mu;mol/L TMP: t=2.056,P＞0.05; 100 mu;mol/L TMP:t=3.058,P＜0.05; 200 mu;mol/L TMP:t=3.828,P＜0.05). ConclusionIn HUVECs, TMP can upregulate the mRNA and protein expression of PHD2, while down regulating HIF-1alpha; protein expression and VEGF mRNA and protein expression under acute hypoxic conditions.
Objective:To observe the intervention effect of the tetra methylpyraz ine on the rds mice with retinitis pigmentosa. Methods:A total of 84 rds mice were randomly divided into 2 groups, with 42 mice in each group. The mice in the experimental group underwent intraperitoneal cavity injection with hydrochlor i c tetramethylpyrazine (80 mg/kg, twice per day) at the date of birth and till 35 days after birth, whereas the normal saline was injected into the intraperito n eal cavity of rats in the control group. The mice were sacrificed 0, 3, 7, 14, 2 1, 28, 35 days after birth, and the eyeballs were enucleated for the routine pat hologic examination with the light microscope. The apoptosis of photoreceptor ce ll nuclei was detected by terminal deoxynucleotidyl transferasemediated dUTP n i ck endlabeling (TUNEL) technigue and the expression of bcl2 in retina was de tect by immunohistochemistry method. Results:The results of li ght microscopy s howed that the layer number of retinal photoreceptor cell nuclei with tetramethy lpyrazine treatment was increased 14, 21, 28, 35 days after the treatment compar ed with that in the control group（P＜0.01）. The results of electron-micro scope suggested that tetramethylpyrazine might reduce lesions in the photoreceptor cells and the destruction of the disc member, mitochondrion，and outer limiting me mbrane in the photoreceptor outer segment in rds mice. The apoptosis of the phot oreceptor cell nuclei reduced in rds mice 3, 7, 14, 21, 28 and 35 days after the treatment compared with that in the control group （P＜0.01）. The express ion of bcl-2 in the matrix of retinal photoreceptor cell nuclei and its inner and o u ter segments increased significantly in rds mice 3，7, 14, 21, 28 and 35 days af ter the treatment （P＜0.05）. Conclusions:Tetramethylpyra zine might reduce ret inal photoreceptor apoptosis by upregulating the expression of bcl-2 in the m at rix of retinal photoreceptor cell nuclei or its inner and outer segments in rds mice.
Objective To provide evidence for clinical practice by assessing the effectiveness and safety of Ligustrazine for primary nephrotic syndrome. Methods We searched MEDLINE (1966.1-2002.12), EMBASE (1975-2002.12), CBM (1979.1-2002.12), Chinese Evidence-Based Medicine/Cochrane Centre Database (CEBM/CCD, Issue 4, 2002) , Cochrane Library, and SCI (1985-2002.12) and handsearched 15 kinds of journals (including Journal of Nephrology et al) (1980-2003.2) for the randomized controlled trials (RCTs).Jadad score was used to assess the quality of RCTs. The outcomes of short term and long term effectiveness, and adverse effect of the treatment were analyzed by RevMan 4.1. Results Thirteen RCTs involving 675 patients met inclusion criteria. Jadad scores of each trial was 1 point. Meta-analysis of 4 studies showed that Ligustrazine had significant better short term effect [OR 4.24, 95% confidence interval (CI) 1.76 to 10.19], lowered 24 h urine protein (OR -0.36, 95% CI -0.71 to -0.02), improved renal function [ creatinine level in children group: (OR -3.34, 95% CI -5.25 to -1.43), creatinine level in adult group: (OR -48.29, 95% CI -68.24 to -28.35)], and increased serum albumin level (OR 3.61, 95% CI 2.61 to 4.61). Whether Ligustrazine had long term side effect was not confirmed. No adequate evidence showed that Ligustrazine could reduce the relapse rate of primary nephrotic syndrome. Conclusions Meta-analysis of low quality RCTs suggest that Ligustrazine does work in primary nephritic syndrome in short term observation. No adequate evidence shows that Ligustrazine has severe side effect or can reduce the relapse rate of primary nephrotic syndrome. We can’t draw a conclusion that Ligustrazine is safe in primary nephrotic syndrome treatment.A rigorously designed, randomized, double blind, placebo controlled trial are required.
Objective To evaluate the efficacy and safety of tetramethylpyrazine (TMP) for diabetic peripheral neuropathy (DPN). Methods Randomized controlled trials were identified from CBM (1978-2003.3), TCMLRS (1980-2003.3), Medline (1970-2003.3), EMbase (1970-2003.3) and Cochrane Library (issue 3, 2003). We handsearched Journal of Traditional Chinese Medicine (1990-2002), New Chinese Medicine (1990-2002), Chinese Journal of Integrated Traditional and Western Medicine (1990-2002) and Research of Traditional Chinese Medicine (1990-2002). Papers of the controlled trials of clinical therapeutic studies on DPN treatment by Chinese medicine herb TMP were included and analyzed according to the criteria of the Cochrane Handbook in 1997. Results Six RCTs involving 669 patients were included, with all trials of low methodological quality. Meta-analysis indicated TMP was more effective than western medicine on pain or numbness of extremities of DPN [The pooled OR = 10.12, 95%CI (6.70 to15.28), P=0.000] and motor nerve conduction velocity change of common peroneal nerves and median nerves . Only one trial reported the side effects of TMP, such as dizziness and headache. Conclusions Based on the review, TMP infusion may have positive effect on DPN. However, the evidence is not b enough due to the general low methodological quality, so we can’t draw a reliable conclusion about the effects of TMP for DPN at the moment. Further large randomized, double blind, placebo-controlled trial are needed.
目的 探讨川芎嗪对大鼠重症急性胰腺炎（SAP）脑损伤的保护作用。方法 将72只健康Wistar大鼠按数字表法随机均分为对照组、SAP组和川芎嗪治疗组3组。对照组仅剖腹翻动胰腺后即缝合腹壁； SAP组采用胰腺被膜下均匀注射5%牛磺胆酸钠（2ml/kg体重）制备SAP动物模型；川芎嗪治疗组在SAP建模后5min于大鼠尾静脉注射川芎嗪注射液（100mg/kg体重）。各组大鼠分别于术后第6、12及24 h观察胰腺组织及脑组织的病理改变，检测血清淀粉酶、 脑组织含水量和微血管内白细胞聚集附壁计数，以及脑组织中MDA、TNF-α和IL-1β水平。结果 对照组胰腺组织无明显改变；SAP组胰腺组织腺泡细胞坏死，结构不清，间质水肿，红细胞漏出，部分腺导管扩张，有点片状出血坏死，炎性细胞浸润；川芎嗪治疗组胰腺组织病理改变较同一时相的SAP组明显减轻。对照组脑组织无明显改变；SAP组脑组织神经元细胞水肿，微血管内白细胞聚集及附壁，脑组织内有炎性细胞增生、聚集，且随时间延长上述表现逐渐加重；川芎嗪治疗组脑组织病理改变较同一时相的SAP组明显减轻。SAP组大鼠各时相脑组织含水量和微血管内白细胞聚集附壁计数，脑组织中TNF-α、IL-1β和MDA水平，以及血淀粉酶含量均明显高于对照组（P＜0.05）；川芎嗪治疗组大鼠各时相的上述指标均明显低于SAP组（P＜0.05）。结论 大鼠脑组织中的TNF-α、IL-1β及MDA参与了SAP脑损伤的病理过程，川芎嗪对SAP大鼠脑损伤具有保护、治疗作用。