Effects of Co-Administration of Icariin and Panax notoginseng Saponins on Intestinal Microbiota and Hippocampal Protein Expression in a Mouse Model of Alzheimer's Disease.

OBJECTIVE: We investigated the effect of icariin (ICA) combined with Panax notoginseng saponins (PNS) on intestinal microbiota and hippocampal protein expression in amyloid precursor protein/presenilin 1 (APP/PS1) transgenic mice, a model of Alzheimer's disease (AD). METHODS: Transgenic mice were treated with icariin and PNS. The Morris water maze (MWM) was used to assess spatial memory, and the gut microbiota and differential protein expression in the hippocampus were investigated using high-throughput screening techniques. Differential protein expression was confirmed by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting. RESULTS: The MWM results showed that the mice treated with the medium dose of ICA+PNS spent significantly more time in the target quadrant compared with the AD group. Bacterial diversity was the lowest in the AD group, with significantly greater diversity in the ICA + PNS treatment group. Three proteins were selected for proteomic analysis, and qRT-PCR and Western blot were used to detect the expression of 2'-5'-oligoadenylate synthetase ubiquitin like 1 (Oasl1), trichoplein keratin filament-binding protein (TCHP), and tumor necrosis factor receptor associated 3-interacting protein 1 (MIPT3). Compared with control mice, MIPT3 expression was increased and Oasl1 and TCHP were reduced in the AD group. These abnormal protein expressions tended to normalization after treatment with medium dose of ICA and PNS. CONCLUSION: Treatment with ICA and PNS ameliorated memory impairment in an AD mouse model. The mechanisms may be related to modulation of the intestinal microbiota and expression of Oasl1, TCHP, and MIPT3.

Tanshinone IIA reduces SW837 colorectal cancer cell viability via the promotion of mitochondrial fission by activating JNK-Mff signaling pathways.

BACKGROUND: Mitochondrial homeostasis has been increasingly viewed as a potential target of cancer therapy, and mitochondrial fission is a novel regulator of mitochondrial function and apoptosis. The aim of our study was to determine the detailed role of mitochondrial fission in SW837 colorectal cancer cell viability, mobility and proliferation. In addition, the current study also investigated the therapeutic impact of Tanshinone IIA (Tan IIA), a type of anticancer adjuvant drug, on cancer mitochondrial homeostasis. RESULTS: The results of our data illustrated that Tan IIA promoted SW837 cell death, impaired cell migration and mediated cancer proliferation arrest in a dose-dependent manner. Functional investigation exhibited that Tan IIA treatment evoked mitochondrial injury, as witnessed by mitochondrial ROS overproduction, mitochondrial potential collapse, antioxidant factor downregulation, mitochondrial pro-apoptotic protein upregulation, and caspase-9-dependent apoptotic pathway activation. Furthermore, we confirmed that Tan IIA mediated mitochondrial damage by activating mitochondrial fission, and the inhibition of mitochondrial fission abrogated the proapoptotic effects of Tan IIA on SW837 cells. To this end, our results demonstrated that Tan IIA modulated mitochondrial fission via the JNK-Mff pathways. The blockade of the JNK-Mff axis inhibited Tan IIA-mediated mitochondrial fission and promoted the survival of SW837 cells. CONCLUSIONS: Altogether, our results identified mitochondrial fission as a new potential target to control cancer viability, mobility and proliferation. Furthermore, our findings demonstrate that Tan IIA is an effective drug to treat colorectal cancer by activating JNK-Mff-mitochondrial fission pathways.

Treating Alzheimer's disease with Yizhijiannao granules by regulating expression of multiple proteins in temporal lobe.

Yizhijiannao granules have been shown to improve cognitive function in Alzheimer's disease patients. The present study sought to explore the mechanisms involved in the cognitive enhancing effects of Yizhijiannao granule. Senescence-accelerated mouse prone 8 mice with learning and memory disorders were intragastrically treated with Yizhijiannao granule for 8 weeks. Mice intragastrically treated with double distilled water for 8 weeks were considered as the control group. 2D gel electrophoresis was used to isolate total protein from the temporal lobe of senescence-accelerated mouse prone 8 mice, and differential protein spots were obtained by mass spectrometry. Thirty-seven differential protein spots were found in the temporal lobe area of both groups. Ten protein spots were identified: high mobility group box 1, dimethylarginine dimethylaminohydrolase-1, neuroglobin, hemoglobin beta adult major chain, peroxiredoxin-6, cofilin-1, flotillin 1, peptidylprolyl isomerase A, voltage-dependent anion channel-2 and chaperonin containing TCP1, and subunit 2. Among other functions, these proteins are separately involved in the regulation of amyloid beta production, oxidative stress, neuroinflammation, regulation of tau phosphorylation, and regulation of neuronal apoptosis. Our results revealed that Yizhijiannao granule can regulate the expression of various proteins in the temporal lobe of senescence-accelerated mouse prone 8 mice, and may be therapeutically beneficial for the treatment of Alzheimer's disease.

Yizhijiannao Granule and a combination of its effective monomers, icariin and Panax notoginseng saponins, inhibit early PC12 cell apoptosis induced by beta-amyloid (25-35).

One of our previous studies showed that Yizhijiannao Granule, a compound Chinese medicine, effectively improved the clinical symptoms of Alzheimer's disease. In the present study, we established a model of Alzheimer's disease using beta-amyloid (25-35) in PC12 cells, and treated the cells with Yizhijiannao Granule and its four monomers, i.e., icariin, catechin, Panax notoginseng saponins, and eleutheroside E. Flow cytometry showed that Yizhijiannao Granule-containing serum, icariin, Panax notoginseng saponins, and icariin + Panax notoginseng saponins were protective against beta-amyloid (25-35)-induced injury in PC12 cells. Icariin in combination with Panax notoginseng saponins significantly inhibited early apoptosis of PC12 cells with beta-amyloid (25-35)-induced injury compared to icariin or Panax notoginseng saponins alone. The effects of icariin + Panax notoginseng saponins were similar to the effects of Yizhijiannao Granule. The findings indicate that two of the effective monomers of Yizhijiannao Granule, icariin and Panax notoginseng saponins, can synergistically inhibit early apoptosis of PC12 cells induced by beta-amyloid (25-35).

Icariin upregulates phosphorylated cyclic adenosine monophosphate response element binding protein levels in the hippocampus of the senescence- accelerated mouse.

At 8 weeks after intragastric administration of icariin to senescence-accelerated mice (P8 strain), Morris water maze results showed that escape latency was shortened, and the number of platform crossings was increased. Immunohistochemical staining and western blot assay detected significantly increased levels of cyclic adenosine monophosphate response element binding protein. These results suggest that icariin upregulates phosphorylated cyclic adenosine monophosphate response element binding protein levels and improves learning and memory functions in hippocampus of the senescence-accelerated mouse.

[Effect of yizhi jiannao granule concentration fluid on the differential expression protein in entorhinal area tissue of senescence accelerated mouse P8].

OBJECTIVE: To explore the mechanism of Yizhi Jiannao Granule (YZJN) in treating Alzheimer's disease (AD) on proteomic level by analyzing the differential expression proteins in entorhinal cortex tissue of senescence accelerated mouse P8 (SAMP8) treated with YZJN. METHODS: Six-month old SAMP8 were randomly divided into 3 groups, the model group, the YZJN group and the control group, 10 mice in each group. The model group was untreated with free water access, the YZJN group was treated with concentrated water extract of YZJN 0.3 g per day via gastric perfusion, and the control group was perfused with equal volume of double distilled water. The total protein in entorhinal cortex tissue of mice was extracted after an 8-week treatment with two-dimensional gel electrophoresis, and the differential expression protein spots were separated for identification through peptide mass fingerprint analysis and database searching. RESULTS: Thirty-two protein spots expressed differentially between the YZJN group and the model group were found, and 14 differential expression proteins were identified, including NADH dehydrogenase iron-sulfur protein 6, Rho-GDP dissociation inhibitor alpha, beta2-globin, phosphoglyceric kinase, etc, their functions involved mitochondria energy metabolism, oxidative stress and neuron function. CONCLUSION: YZJN could regulate multiple protein expressions in entorhinal cortex tissues of SAMP8, suggesting that it has multi-target therapeutic action and its mechanism in treating AD is possibly realized by way of improving mitochondria function, antagonizing oxidation stress, preventing nerve cell apoptosis and protecting neurons.

[Influence of acupuncture on isoprostane in patients with Alzheimer's disease].

OBJECTIVE: To explore the clinical therapeutic effect of acupuncture on Alzheimer's disease (AD) and its mechanism. METHODS: Twenty patients with Alzheimer's disease were treated by acupuncture with reinforcing kidney and activating blood method for 12 weeks and Baihui (GV 20), Shenshu (BL 23), Xuehai (SP 10) and Geshu (BL 17) were selected. The clinical therapeutic effect were assessed by comparing the scores of Alzheimer's Disease Assessment Scale-Cognitive Section (ADAS-Cog) and 8-IPF2alpha concentration in cerebrospinal fluid, blood and urine before and after treatment were detected by using enzyme linked immunosorbent assay. RESULTS: After treatment, the effective rate was 90.0%. The score of ADAS-Cog was 35. 70 +/- 14. 70 before treatment and 31. 45 +/- 4. 08 after treatment, with a significant difference (P<0. 001). The concentration of 8-IPF2alpha in cerebrospinal fluid, blood and urine were all significantly decreased after treatment (all P<0.001). CONCLUSION: Acupuncture can improve the cognitive ability of AD patients and its possible mechanism may be relative to the decrease in lipid peroxidation in AD patients' brain.

[Clinical observation on acupuncture combined with Yizhi Jiannao granules for treatment of Alzheimer's disease].

OBJECTIVE: To observe clinical therapeutic effect of acupuncture combined with Yizhi Jiannao Granules for treatment of Alzheimer's disease and its effects on intelligence, daily life and social activity ability. METHODS: Eighty-four cases were randomly divided into 3 groups, 28 cases in each group. The combined acupuncture and medication group was treated with acupuncture at Baihui (GV 20), Sishencong (EX-HN 1), Dazhui (GV 14), Guanyuan (CV 4), etc. and oral administration of Yizhi Jiannao Granules; the Chinese herb group was treated with Yizhi Jiannao Granules, and the western medicine group with oral administration of Aricept. The scores for the Mini-Mental State Examination (MMSE), Ability of Daily Life (ADL) and the therapeutic effects were assessed and compared before treatment and after treatment for 12 weeks among the groups. RESULTS: After treatment, the scores for MMSE and ADL were improved in the combined acupuncture and medication group, the Chinese herb group and the western medicine group, which were better in the combined acupuncture and medication group (P < 0.05). The total effective rate of 85.7% in the combined acupuncture and medication group was better than 71.4% in the Chinese herb group and 67.9% in the western medicine group. CONCLUSION: Acupuncture combined with Yizhi Jiannao Granules has a significant therapeutic effect on Alzheimer's disease, which is better than that of Yizhi Jiannao Granules or Aricept.

[Effect of yizhi jiannao granules on the expression of Pin1 and HMGB1 mRNA in the hippocampus of SAMP8 mice].

OBJECTIVE: To investigate the effect of yizhi jiannao granule concentration fluid (YCF) on the expression of peptidyl-prolyl-cis-trans isomerase A (Pin1) and high mobility group box 1 (HMGB1) mRNA in the hippocampus of senescence accelerated mice Senile-Prone8(SAMP8). METHODS: Six-month old SAMP8 mice were randomly divided into a YCF group and a model group. Six-month old SAMP8 mice were served as a normal control group. The YCF group was ravaged, while the model group and the normal control group were gavaged with double-distilled water for 8 weeks. The hippocampus was taken out for examination. The expression of Pin1 and HMGB1 mRNA was measured by RT-PCR. RESULTS: In the YCF group, the level of Pin1 mRNA increased, and the level of HMGB1 mRNA decreased compared with that of the model group. CONCLUSION: Yizhi jiannao granules can prevent Alzheimer's disease by increasing the Pin1 level and decreasing the HMGB1 level.

[Effect of compound Rhizoma Smilacinus granules on the expression of Bcl-2 and Bax gene in A549 cell lines of non-small cell lung cancers].

OBJECTIVE: To determine the effect of compound Rhizoma Smilacinus granules (CRSG) on the expression of Bcl-2 and Bax gene in A549 cell lines, and to explore the mechanism of CRSG on non-small cell lung cancers. METHODS: The SD rats were randomly divided into 2 groups: one was administrated with CRSG (n=15), and the other with the same dose of distilled water (n=15). The herbage-containing serum was obtained 2 hours after the 6th treatment. non-small lung cancer A549 cell lines were randomly divided into CRSG group, diamminedichloroplatinum (DDP) group and normal control group, which were cultivated with 10% herbage-containing serum, DDP (3g/mL), and 10% SD rats serum respectively. The cultivated cells were collected after 48 hours, and then RT-PCR technique was used to determine the expression of Bcl-2 and Bax mRNA. RESULTS: Compared with the normal control group after 48 h, the level of Bcl-2 mRNA and the rate of Bc-2/Bax decreased in the CRSG group and the DDP group, and the level of Bax mRNA increased with significant difference (P<0.01). CONCLUSION: non-small cell lung cancer A549 cell lines have a high level of Bcl-2 mRNA and a low level of Bax mRNA. CRSG can significantly down-regulate the expression of Bcl-2 mRNA and the rate of Bc-2/Bax, and obviously up-regulate the expression of Bax mRNA, which probably is one of the molecular mechanisms of CRSG in inhibiting the growth of non-small cell lung cancers.

[Effect of yizhi jiannao granule on the behavior and neuron apoptosis in SAMP/8 mice].

OBJECTIVE: To investigate the effect of yizhi jiannao granule concentration fluid (YCF) on the behavior, the apoptosis rate of hippocampus neuron and the expression of apoptosis gene Bcl-2, Bax in senescence accelerated mice Senile-Prone/8 (SAMP/8), and to discuss some mechanism of traditional chinese medicine YCF in improving the capability of learning and memory. METHODS: Forty 6-month old SAMP/8 mice were randomly divided into the old group, huperzine A (Hup-A) group and YCF group. Ten 4-month old SAMP/8 mice were served as a young control group. Four groups were given different drugs for 8 weeks, their behavior changes were observed, and the hippocampus were taken out to examine the apotosis rate by flow cytomeutry (FCM) and the expression of Bcl-2, Bax mRNA by RT-PCR. RESULTS: In the YCF group, the escape latency was significantly shortened, the time of swim in the platform quadrant significantly increased, the apoptosis rate of hippocampus neural decreased; the level of Bcl-2 mRNA and the rate of Bcl-2/Bax increased, and the level of Bax mRNA decreased. CONCLUSION: Yizhi jiannao granule can decrease the neuron apoptosis rate and the Bax level, increase the Bcl-2 level, and modulate the rate of Bcl-2/Bax in SAMP/8 brain, which is probably part of the mechanisms of inhibiting the apoptosis and improving learning and memory.

[Effect of xiaokeling concentration fluid on mRNA expression of insulin-like growth factor-1 in sciatic nerve of Streptozotocin-induced diabetic rats].

OBJECTIVE: To investigate the effect of xiaokeling concentration fluid on insulin-like growth factor-1 (IGF-1) mRNA expression in sciatic nerve of Streptozotocin-induced diabetic rats. METHODS: Thirty diabetic rats were randomly divided into model group, mecobalamin tablets group, and xiaokeling concentration fluid group. The IGF-1 mRNA level in sciatic nerve of each group was determined after 8 weeks by relative quantity RT-PCR. RESULTS: The IGF-1 mRNA level in sciatic nerve of diabetic rats between xiaokeling concentration fluid group, mecobalamin tablets group and normal group showed no significant difference ( P = 0.213, P = 0.822, P = 0.304 ), while was significantly higher than that of the model group ( P < 0.05 ). IGF-1 mRNA level was negatively correlated with the level of blood sugar (P < 0.05). CONCLUSION: IGF-1 mRNA level decreased in sciatic nerve of diabetic rats. Xiaokeling concentration fluid can increase the IGF-1 mRNA level in sciatic nerve of diabetic rats. Xiaokeling concentration fluid is involved in the regulation of IGF-1 expression, and probably prevents diabetic peripheral neuropathy from deteriorating.