20(S)-protopanaxadiol induces apoptosis in human umbilical vein endothelial cells by activating the PERK-eIF2alpha-ATF4 signaling pathway.

20(S)-protopanaxadiol (PPD)-type ginsenosides are generally believed to be the most pharmacologically active components of Panax ginseng. These compounds induce apoptotic cell death in various cancer cells, which suggests that they have anti-cancer activity. Anti-angiogenesis is a promising therapeutic approach for controlling angiogenesis-related diseases such as malignant tumors, age-related macular degeneration, and atherosclerosis. Studies showed that 20(S)-PPD at low concentrations induces endothelial cell growth, but in our present study, we found 20(S)-PPD at high concentrations inhibited cell growth and mediated apoptosis in human umbilical vein endothelial cells (HUVECs). The mechanism by which high concentrations of 20(S)-PPD mediate endothelial cell apoptosis remains elusive. The present current study investigated how 20(S)-PPD induces apoptosis in HUVECs for the first time. We found that caspase-9 and its downstream caspase, caspase-3, were cleaved into their active forms after 20(S)-PPD treatment. Treatment with 20(S)-PPD decreased the level of Bcl-2 expression but did not change the level of Bax expression. 20(S)-PPD induced endoplasmic reticulum stress in HUVECs and stimulated UPR signaling, initiated by protein kinase R-like endoplasmic reticulum kinase (PERK) activation. Total protein expression and ATF4 nuclear import were increased, and CEBP-homologous protein (CHOP) expression increased after treatment with 20(S)-PPD. Furthermore, siRNA-mediated knockdown of PERK or ATF4 inhibited the induction of CHOP expression and 20(s)-PPD-induced apoptosis. Collectively, our findings show that 20(S)-PPD inhibits HUVEC growth by inducing apoptosis and that ATF4 expression activated by the PERK-eIF2α signaling pathway is essential for this process. These findings suggest that high concentrations of 20(S)-PPD could be used to treat angiogenesis-related diseases.

[Effect of tianma gouteng decoction on the endothelial function and the renal protein expression in spontaneously hypertensive rats].

OJECTIVE: To observe the effect of tianma gouteng decoction (TGD) on the endothelial function and the renal protein expression of spontaneously hypertensive rats, and to analyze its possible mechanism. METHODS: Totally 18 6-week-old SHR were randomly divided into 3 groups according to randomized block design, the SHR control group, the TGD group, and the captopril group, 6 in each group. Meanwhile, Wistar Kyoto (WKY) rats of the same age were recruited as a WKY control group. Rats in the TGD group were administered with TGD at the daily dose of 10. 260 g/kg. Rats in the captopril group were administered with captopril at the daily dose of 3. 375 g/kg. 2 mL/100 g distilled water was administered to rats in the SHR control group and the WKY control group. All medication was performed by gastrogavage once per day till rats were 24 weeks old. Changes of blood pressure were measured once per two weeks. The relaxation of the thoracic aorta and the superior mesenteric artery was determined by vascular ring in vitro to reflect the endothelial function. The total renal protein was separated by two-dimensional electrophoresis (2-DE). The significantly deviated protein was verified by Western blot. RESULTS: (1) Compared with the SHR control group, blood pressure was significantly lowered in rats (10 - 24 weeks old) of the captopril group (P <0.01, P <0.05). The hypotensive effect of TGD was obvious at the beginning of hypertension (10 -12 weeks) (P <0. 01). But along with the progression of hypertension, its hypotensive effect was not obvious (P>0. 05). (2) Compared with the SHR control group, the relaxation of the superior mesenteric artery was obviously improved in the TGD group (P <0. 05); the relaxation of the thoracic aorta and the superior mesenteric artery was obviously superior in the WKY control group (P <0. 01, P <0. 05). But there was no statistical difference in each relaxation index between the captopril group and the SHR control group (P >0. 05).(3) RESULTS: of 2-DE found 16 significantly differential renal protein, mainly involved nitric oxide (NO) system, oxidative stress, and cytoskeleton-related proteins. Results of Western blot showed that TGD could significantly improve expressions of Cu-Zn superoxide dismutase (SOD), N(G, N(G)-dimethylarginine dimethylaminohydrolase 2 (DDAH2), and pterin-4-alpha-carbinolamine dehydratase 1 (PCBD1) (P <0. 05). CONCLUSION: GTD could protect the endothelial function of the superior mesenteric artery in SHR, and its intervention mechanism of hypertension induced early renal injury might be relevant to regulating the NO system and antioxidative stress.

[Analysis of renal protein expressions of spontaneously hypertensive rat intervention by three different Chinese medicine compounds for hypertension].

OBJECTIVE: To observe the differential protein expression profile of three different Chinese medicine compounds on spontaneous hypertensive rat, which were Banxiabaizhutianma Decoction, Dihuangyinzi Decoction and Tianmagouteng Decoction, and to elaborate their function characters and regulation pathway using the principal component analysis and analyze the basic theory of different treatment of hypertension. METHODS: Give three compounds to 6-week-old SHR (Spontaneous hypertensive rat) as experimental group and captopril to 6-week-old WKY (Wistar-Kyoto rat) as control group by gavage. Monitor the blood pressure of rat tail to 24-week-old and then all rats were decapitated. Extracte the rental protein to finish two-dimensional electrophoresis experiments and get the differential protein spots using PDQuest software. Study the three different compounds' effect of trend on SHR blood pressure using PCAC principal component analysis. RESULTS: The blood pressure of SHR remained rising from 6-week-old to 10-week-old, when Tianmagouteng Decoction had a significant anti-hypertension effect. But as the blood pressure of SHR remained high, Banxiabaizhutianma Decoction and Dihuangyinzi Decoction performed better in lowering blood pressure. At last, renal protein expression profile analysis showed that there were 84 significantly different proteins between SHR and WKY of 24-week-old and the PCA result showed that the principal component extraction was 95.951%, it was sure that Tianmagouteng Decoction and Banxiabaizhutianma Decoction had significantly changed rental protein spots on SHR while Dihuangyinzi Decoction had not. CONCLUSION: In the early stages of rising blood pressure, Tianmagouteng Decoction has significant anti-hypertensive effect. But when the blood pressure remain high, Banxiabaizhutianma Decoction and Dihuangyinzi Decoction are superior to Tanmagouteng Decoction in anti-hypertension. Banxiabaizhutianma Decoction and Tianmagouteng Decoction can significantly improve rental protein spots on SHR while Dihuangyinzi Decoction can not.

[The effect of Banxia Baizhu Tianma decoction on renal protein expression in spontaneously hypertensive rats].

OBJECTIVE: To study the effect of Banxia Baizhu decoction on the spontaneously hypertensive rats and its possible mechanism. METHODS: Male SHRs of 6 weeks old were randomly and equally divided into two groups, the control SHR group and the group treated with BXBZTMD, the treated group were treated with BXBZTMD orally for 18 weeks, the control SHR group and normotensive WKY group were fed with the same amount of distilled water. Rats' blood pressure was measured through carotid artery catheterization, and protein of kidney tissues was analyzed by two-dimensional electrophoresis after extraction and purification. Different protein spots expressed significantly which were anlyzed by PDQuest software were then identified by MALDI-TOF/TOF MS. The protein expressions of peroxidase 2 and Cu-Zn superoxide in kidney were determined by Western blot. RESULTS: Compared SHR with WKY, 12 protein spots were changed. BXBZTMD could adjust kidney protein expression such as Cu-Zn superoxide dismutase, peroxidase 2 and may change kidney protein expression such as the alpha-beta-crystallin, which were closely related to oxidative stress. These discoveries of the target proteins suggested that BXBZTMD had curative effect by anti-oxidative stress. CONCLUSION: Oxidative stress plays an important role in the process of hypertensive renal injury. BXBZTMD can play a therapeutic role to oxidative stress. The discovery of the target protein provides a theoretical basis for the mechanism of the therapeutic effect of BXBZTMD.

[Effect of banxia baizhu tianma decoction on the vascular endothelial function of spontaneous hypertensive rats].

OBJECTIVE: To study the vascular endothelial function recovery and its mechanism of Banxia Baizhu Tianma Decoction (BBTD). METHODS: 54 SH rats were randomly divided into three groups: the blank control group, BBTD group and Captopril treatment group. BBTD (at the daily dose of 4. 320 g crude drug/kg) and Captopril (at the daily dose of 3.375 g/kg) was administered from the 7th week to the 24th week. Another eighteen Wistar-Kyoto rats of the same ages were taken as the control. Medication was discontinued and effects were observed until the 32nd week. The blood pressure was determined by arterial carotis cannula. The concentration of serum NO2(-) and total anti-oxidation were determined by Griess and fluorescence recovery after photobleaching (FRAP). The acetylcholine (Ach)-dependent relaxation of superior mesenteric artery was detected using in vitro vascular ring. The mRNA expressions of IL-1, IL-6 and iNOS were detected by Real-time PCR at the 18th, 24th, and 32nd week. RESULTS: BBTD could significantly lower blood pressure of SHR and the concentration of serum NO2(-) at the 18th and 24th week (P<0.05). The total anti-oxidation of SH rats increased at the 18th week (P<0.01), and ACh-dependent relaxation of superior mesenteric artery increased at the 24th week. The mRNA expressions of IL-1 was markedly suppressed by BBTD at the 18th, 24th, and 32nd week (P<0.05), while IL-6 and iNOS mRNA expression were significantly lowered only at the 32nd week (P< 0.01). Captopril could significantly lower blood pressure of SHR at the 18th and 24th week (P<0.05). It significantly increased the total anti-oxidation of SH rats at the 18th week (P<0.01). However, it could not increase ACh-dependent relaxation of superior mesenteric artery and regulate the concentration of NO2(-) at the 18th, 24th, and 32nd week. The mRNA expression of iNOS was markedly suppressed by Captopril at the 24th and 32nd week, while mRNA expressions of IL-1 and IL-6 were significantly lower only at the 32nd week (P<0.01). CONCLUSIONS: BBTD showed similar effect in decreasing the blood pressure to captopril, but it showed better effect in improving the mesenteric endothelial dysfunction of SHR, which may be associated with its inhibition on NO and IL-1 expression, and improvement of the oxidative stress state.

[Effect of banxia baizhu tianma decoction on the left ventricular hypertrophy of hypertrophied myocardium in spontaneously hypertensive rat].

OBJECTIVE: To investigate the changes in renin angiotensin system (RAS) in hypertrophied myocardium of spontaneous hypertensive rat (SHR), and the effect of Banxia Baizhu Tianma Decoction (BBTD) on the changes in haemodynamic parameters and mRNA of signaling molecules of RAS at different periods. METHODS: Fifty-four male SHRs of 6 weeks old were randomly and equally divided into three groups: the untreated control group, the test group, and the positive control group, and they were treated respectively with distilled water, BBTD and captopril by dissolving in equal volume of water administrated via gavage for 18 weeks. Besides, 18 age matched Wistar-Kyoto (WKY) rats treated with distilled water were allocated in a normal control group. Rats were managed in batches at their age of 18, 24, and 32 weeks old. Rat's hemodynamic parameters were measured through carotid artery catheterization, myocardial pathology was observed, and their mRNA expressions of angiotensin (AGT), angiotensin-converting (ACE) and angiotension-converting 2 (ACE2) were determined by Real-time PCR. RESULTS: Compared with WKY rats, the arterial pressure and left ventricular mass index (LVMI)in SHR were significantly higher at 18, 24 and 32 weeks respectively (P < 0.01); average cycle rate showed in electrocardiogram was higher (P < 0.05), though the blood stream was similar; mRNA expressions of AGT and ACE in heart tissue were markedly higher (P < 0.01), but that of ACE2 at 18 and 24 weeks were lower (P < 0.01). Compared with untreated SHR, arterial pressure at 18 and 24 weeks was lower (P < 0.05); cardiac muscle structure was improved; LVMI at 24 weeks was improved (P < 0.05); the mRNA expressions of AGT and ACE were suppressed but that of ACE2 increased at 18, 24,and 32 weeks significantly in the test group after BBTD treatment (P < 0.05). CONCLUSIONS: Changes in RAS in the hypertrophied myocardium of SHR may be one of the molecular mechanisms for hypertension leading to left ventricular hypertrophy. BBTD can improve the hemodynamic parameters, regulate RAS, so as to lower the arterial pressure.

[Effect of total saponins from rhizomes and flowers of Panax notoginseng on tumor cell induced platelet aggregation].

OBJECTIVE: To investigate the effects of total saponins from rhizomes and flowers of Panax notoginseng on tumor cell induced platelet aggregation (TCIPA). METHODS: MDA-MB-231 breast carcinoma was applied as inductor and the platelet aggregation were investigated by Born's method in vitro and in vivo. RESULTS: Saponins from the flowers of Panax notoginseng inhibited TCIPA at the dose of 120, 240, 480 mg/L respectively in vitro and in vivo. Saponins from rhizomes of Panax notoginseng inhibited TCIPA only at its high dose in vivo. CONCLUSION: The saponins from Panax notoginseng has a dose-dependent inhibition of TCIPA. It may be a new class of antimetastatic agent.