[Research progress on in vitro models of cardiomyocyte injury].

Cardiovascular diseases seriously endanger human health and life. The accompanying myocardial injury has been a focus of attention in society. Chinese medicine,serving as a natural and precious reservoir for the research and development of new drugs,is advantageous in resisting myocardial injury due to its multi-component,multi-pathway,and multi-target characteristics. In recent years,with the extensive application of culture method for isolated cardiomyocytes,a cost-effective,controllable in vitro model of cardiomyocyte injury with uniform samples is becoming a key tool for mechanism research on cardiomyocyte injury and drug development.A good in vitro model can reduce experimental and manpower cost,and also accurately stimulate clinical changes to reveal the mechanism. Therefore,the selection and establishment of in vitro model are crucial for the in-depth research. This study summarized the modeling principles,evaluation indicators,and application of more than ten models reflecting different clinical conditions,such as injuries induced by hypoxia-reoxygenation,hypertrophy,oxidative stress,inflammation,internal environmental disturbance,and toxicity. Furthermore,we analyzed advantages and technical difficulties,aiming to provide a reference for in-depth research on myocardial injury mechanism and drug development.

[Effect of total phenolic part of Rhus chinensis against myocardial ischemia in mice].

Rhus chinensis is an important resource plant. The aqueous extract of R. chinensis roots or stems was to produce Shuguantong Syrup, which is mainly used for the treatment of coronary heart disease and angina pectoris with definite curative effect. On this basis, the crude phenolic part of R. chinensis prepared by macroporous resin was evaluated for the cardio protective effect against myocardial ischemia in mice. The results showed that the phenolic part group with oral administration at the dosages of 190.8-381.6 mg·kg~(-1), compared with the model group, reduced the values of left ventricular end systolic diameter(LVEDs) and the left ventricular end diastolic diameter(LVEDd), and increased the cardiac ejection fraction(EF) and left ventricular fractional shortening(FS) rate, which could effectively improve cardiac function and exert its anti-myocardial ischemia effect, and reduce the rising levels of creatine kinase isoenzyme(CK-MB) and lactate dehydrogenase(LDH) in serum. HE staining showed that the phenolic part group reduced the infiltration of myocardial inflammatory cells and alleviated the degree of myocardial fibrosis and collagen deposition. TUNEL staining showed that the blue-green fluorescence of the phenolic part group decreased successively, and the degree of myocardial cell apoptosis was reduced. Immunohistochemical staining suggested that it could reduce the number of positive cells for p53 protein expression and significantly improve myocardial cell damage. All above data suggested that the phenolic part group had an anti-mycardial ischemis effect. Related mechanism studies revealed that the crude phenolic part could regulate the expressions of the p53 gene(p53), Bcl-2-associated X protein(Bax), B lymphoma-2 gene(Bcl-2), and caspase-3 protein(caspase-3) in myocardial tissue, suggesting that it could reduce cardiac remodeling and myocardial ischemic damage, and improve cardiac function by inhibiting myocardial apoptosis.This research laid a foundation for the elucidation of the pharmacological ingredients R. chinensis.

[Advances on anti-tumor mechanisms of zerumbone].

Zerumbone(ZER), one of humulane-type sesquiterpenoids, showed its unique advantage against tumor activities. The main underlying mechanisms include inhibiting the growth and proliferation of cancer cells, inducing apoptosis of cancer cells and differentiation of cancer cells, regulating immune function, inhibiting invasion and metastasis of cancer cells, and reversing multidrug resistance of cancer cells. Studies on ZER focusing its cytotoxic or anti-tumor is one of hot topic. Currently, with the increasing studies on ZER, the clarified mechanisms are getting rich. The present paper describes a summary of its anti-tumor mechanism of action and helps to provide significant reference to more in-depth research.

[Chemical and pharmacological progress on a Tibetan folk medicine formula Bawei Chenxiang Powder].

Bawei Chenxiang Powder is a traditional Tibetan folk medicine formula, consisting of resinous wood of Aquilaria sinensis, kernel of Myristica fragrans, fruit of Choerospondias axillaris, travertine, resin of Boswellia carterii or B. bhaw-dajiana, stem of Aucklandia lappa, fruit of Terminalia chebula(roasted), and flower of Gossampinus malabarica. It has the function of clearing heart heat, nourishing heart, tranquilizing mind, and inducing resuscitation, which has been used for the treatment of coronary heart disease and angina pectoris. Modern research shows that the medicine materials of this formula mainly contain terpenoids like sesquiterpenes and triterpenes and polyphenols like flavonoids, lignans, and tannins, displaying some pharmacological activities such as anti-myocardial ischemia, anti-cerebral ischemia, and spatial learning and memory promotion. This review summaries the traditional uses, chemical constituents, and pharmacological activities research progress, hopefully to provide a reference for clarification of its pharmacological active ingredients.

[Analgesic and sedative effects of Mongolian medicine Syringa pinnatifolia].

The peeled root,stem or twig of Syringa pinnatifolia is a representative Mongolian folk medicine with the effects of antidepression and pain relief. It has been used for the treatments of heart tingling,heart palpitations,upset,insomnia and other symptoms. Inspired by Mongolian medical theory and clinical practices,this study evaluated the analgesic effect of S. pinnatifolia ethanol extract( T) through three analgesic models including acetic acid writhing test,formalin test,and hot plate test,and the sedative effect of T was evaluated by locomotor activity and synergistic sleeping experiments,and furthermore the effects of T on the GABAergic nervous system were investigated by ELISA,immunohistochemistry,Western blot,and PCR methods. The results showed that T can significantly reduce the number of writhing,the time of paw licking and extend the thermal threshold of mice,suggesting the analgesic effect of T.T also can indicate its sedative effect by reducing the number of activities,decreasing latency of sleeping and extending sleeping time of mice. ELISA results showed that T can increase the content of GABA/Glu in rat cortex,hippocampus,and hypothalamus,and the most significant increase in hypothalamus. The immunohistochemistry and Western blot results showed that T can up-regulate the expression of GAD67 protein in hypothalamus,and the PCR results showed that T can up-regulate the expression of GABAA Rα1,α2,α3,α5,ß1-3,γ1-3 genes,suggesting a sedative effect through the GABAergic nervous system. In conclusion,this study shed insight into the theoretical basis and clinical application of S. pinnatifolia,and also provides inspiration for subsequent development and application.

[Distribution of traditional Chinese medicine syndrome types of male infertility in Chengdu area].

OBJECTIVE: To explore the distribution characteristics of the traditional Chinese medicine (TCM) syndrome types of male infertility in Chengdu area, China, in order to provide some objective evidence for the clinical treatment and studies of male infertility. METHODS: We collected the clinical data on 500 cases of male infertility treated in the outpatient and inpatient departments of the Affiliated Hospital of Chengdu University of Chinese Medicine and Chengdu Hospital for Reproduction, Women and Children from January to December 2017. Based on the results of examinations using the four diagnostic methods of TCM, namely, observation, auscultation and olfaction, interrogation, and palpation, we differentiated the TCM syndromes of the patients and analyzed the distribution characteristics of the syndrome types. RESULTS: Analysis of the baseline characteristics and the results of Chi-square test showed statistically significant differences in the distribution of the frequency of sexual intercourse, body mass index, history of urinary tract infection, testis volume, testicular texture, vas deferens and varicocele among the 500 patients (P < 0.05). As for the distribution of the TCM syndrome types, 115 cases (23.0%) were diagnosed with the unlicensed discernible type of syndrome, 109 (21.8%) with kidney-yang deficiency, 36 (7.2%) with kidney-essence deficiency, 30 (6.0%) with both kidney-yang deficiency and liver-qi stagnation, 30 (6.0%) with both kidney-yang deficiency and spleen-asthenia with excessive dampness, 28 (5.6%) with kidney-yin deficiency, 20 (4.0%) with spleen-asthenia and excessive dampness, 19 (3.8%) with liver-qi stagnation, 19 (3.8%) with phlegm dampness obstruction, 19 (3.8%) with kidney-yang and kidney-essence deficiency, 16 (3.2%) with downward damp-heat, 11 (2.2%) with both kidney-yin and kidney-yang deficiency, 10 (2.0%) with qi-stagnation and blood stasis, and 38 (7.6%) with other types of syndromes. CONCLUSIONS: The main TCM syndrome types of male infertility in Chengdu area include kidney-yang deficiency, kidney-yang deficiency with liver-qi stagnation, and kidney-yang deficiency with spleen-asthenia and excessive dampness. The distribution and influencing factors of the syndrome types need to be further explored and clarified by more large-sample and high-quality studies.

[Pharmacological evaluation of Mongolian medicine Syringa pinnatifolia fraction I against acute myocardial ischemia in mice].

Syringa pinnatifolia Hemsl.( SP) is a representative Mongolian folk medicine with the effects of inhibiting Heyi related diseases,clearing heat and relieving pain. It has been used for the treatment of Heyi-induced heart tingling,heart palpitations,upset,insomnia and other symptoms. Total ethanol extract( T) and major fraction( M) of SP have been evaluated its anti-ischemic effects,and the mechanism was related to the regulation of cyclooxygenase( COX)-mediated inflammatory pathway and p53-mediated apoptosis pathway in our previous studies. This study reports the chemical fractionation on M by which to obtain subfractions( I and M_3),and the pharmacological evaluation of M,I,and M_3 against myocardial ischemia in mice. The result showed that I and M reduced the values of LVEDd and LVEDs,significantly increased EF and FS values,increased serum CK-MB and LDH levels in mice,and reduced in inflammatory cells infiltration and collagen deposition in the infarcted myocardial tissue,suggesting that M and I possess the same degree anti-myocardial is chemia equally whereas M_3 has no this effect. Related mechanism studies suggested that I can reduce the expression of COX-1,COX-2 and p53 protein in myocardial tissue in a dose-dependent manner. This study lays the foundation for further chemical segmentation and clarification of pharmacological substance groups,paving the way for the full use and benefits to be use of systematic biological methods to analyze the pharmacological basis of SP against myocardial ischemia.

[Phytochemical and pharmacological progress on humulane-type sesquiterpenoids].

Humulane-type sesquiterpenoids, widely distributed in plants and microbes, include three types: α-humulene, ß-humulene, and γ-humulene. Up to now, 98 humulane-type sesquiterpenoids have been reported, which possessed anti-tumor, anti-inflammatory, antibacterial, and antiviral activities. Herein, this paper describes their chemical constituents and pharmacological activities, hoping to bring benefits for further research and lay a foundation for investigating the structure-activity relationships.

[Inhibitory effect of Qilan Capsules on the expressions of vasculogenic mimicry-related proteins in prostate cancer].

Objective: To investigate the effect of Qilan Capsules (QLC) on the expressions of the related proteins HIF-1α, VEGF-α, EphA2 and MMP-1 in the formation of vasculogenic mimicry (VM) in prostate cancer. METHODS: Prostate cancer PC-3 cells were cultured, transfected with siRNA, and divided into eight groups, blank control, HIF-1α siRNA, VEGF-α siRNA, EphA2 siRNA, QLC intervention, QLC + HIF-1α siRNA, QLC + VEGF-α siRNA, and QLC + EphA2 siRNA. The expressions of the HIF-1α, VEGF-α and EphA2 proteins in the pathway of VEGF were determined by Western blot. RESULTS: Compared with the blank control group, the expression of HIF-1α was evidently decreased in the HIF-lα siRNA and QLC + HIF-lα siRNA groups (0.624 7 ± 0.042 8 vs 0.032 8 ± 0.002 5 and 0.036 8 ± 0.018 1, P < 0.05), so were that of VEGF-α in the VEGF-α siRNA and QLC + VEGF-α siRNA groups (0.068 9 ± 0.005 1 vs 0.016 9 ± 0.000 7 and 0.010 9 ± 0.000 8, P < 0.05), that of EphA2 in the EphA2 siRNA and QLC + EphA2 siRNA groups though with no statistically significant difference (0.1684 ± 0.0126 vs 0.134 5 ± 0.028 6 and 0.165 4 ± 0.039 8, P > 0.05), and that of MMP-1 in the HIF-lα siRNA, VEGF-α siRNA and EphA2 siRNA groups (1.696 1 ± 0.152 7 vs 0.435 9 ± 0.036 9, 0.198 7 ± 0.009 0 and 0.0218 ± 0.000 7, P < 0.05). CONCLUSIONS: Qilan Capsules can suppress VM formation in prostate cancer by inhibiting the expressions of HIF-1α, VEGF-α and MMP-1, which plays a role in the clinical treatment of prostate cancer by checking the growth and development of the blood supply system in the tumor tissue.

[Chemical constituents from a Tibetan herbal medicine Corydalis hendersonii].

Nine alkaloids and two phenolic glycosides were isolated from EtOH extract of the whole plants of Corydalis hendersonii by various chromatographic techniques including silica gel, ODS, Sephadex LH-20, and semi-preparative HPLC. Their structures were identified as groenlandicine (1), berberine (2), protopine (3), cryptopine (4), N-trans-feruloyloctopamine(5), 3-(4-hydroxy-3-methoxyphenyl)-N-[2-(4-hydroxyphenyl)-2-methoxyethyl] acrylamide (6), N-cis-p-coumaroyloctopamine (7), N-trans-p-coumaroylnoradrenline (8),N-cis-feruloyloctopamine (9), apocynin (10), and glucoacetosyringone (11) by the spectroscopic data analysis and comparison with those in the literature. Among them, compounds 10 and 11 were isolated from this genus for the first time, and 1, 2, and 5-9 were isolated from the species for the first time. All isolates were tested for their protection for in vitro PC12 cell line and antiplatelet aggregation activity. The results showed that compounds 5 and 7 displayed protective effects at a concentration of 10 µmol·L⁻¹, and compound 2 showed antiplatelet aggregation activity induced by THR, ADP, and AA, and compound 3 exhibted inhibitory effect induced by THR.

[Effects of Scutellaria Barbata on VEGF Expression in K562 Cells].

OBJECTIVE: To investigate the anti-angiogenesis effect of Scutellaria barbata extract(SBE) on chronic myeloid leukemia(CML) K562 cell line in vitro. METHODS: The proliferating activity after treating K562 cells with 0.5,1.0,2.0 and 4.0 g/ml SB for 24, 36, 48 hours were assessed by MTT assay. The level of vascular endothelial growth factor(VEGF) in the culture supematant of K562 cells was determined by ELISA; and the expression of VEGF mRNA was detected by RT-PCR. RESULTS: MTT assay showed that SBE could inhibit the proliferation of K562 cells in a dose-dependent manner (r=0.56); ELISA displayed that the concentration of VEGF in K562 cells in blank-control group was most high; after intervention of K562 cells by SBE (0.5,1.0,2.0 and 4.0 g/ml) for 48 h, the concentration of VEGF decreased, the comparison between different groups showed significant differences (P<0.05); after treatment with SBE for 48 h, the expression of VEGF mRNA in K562 cells decreased, the gray scale ratio of target gene/ß-actin declined, and the difference between various groups was statistically significant (P<0.05). Conclution: SBE can inhibit K562 cell proliferation, its action mechanism may related with the VEGF level concentration in K562 cells and down-regulation of VEGF mRNA expression.

[Inhibitory Effect of Serum Containing Fuzheng Jiedu Decoction on the Leukemia K562/A02 Multi-drug Resistance Cells and Its Mechanism].

OBJECTIVE: To study the inhibitory effect of serum containing Fuzheng Jiedu decoction on leukemia multi-drug-resistance K562/A02 cells and its possible mechanism. METHODS: The MTT method was used to detect the inhibitory rate of K562/AO2 cells treated with serum containing Fuzheng Jiedu decoction; the flow cytometry was used to detect the inhibitory effect of serum containing medicin on growth of K562/AO2 cells and P-gp expression; the Q-PCR was used to assay the BCL-2 mRNA expression; the Western blot was used to detect the BCL-2 protein expression. RESULTS: MTT cytotoxic test showed serum containing Fuzheng Jiedu decoction could inhibit K562/A02 cell growth, and the inhibitory rate increased with the increase of drug concentration; the flow cytometry showed that the serum containing Fuzheng Jiedu decoction could promote K562/A02 cell apoptosis in a concentration-dependent manner. qPCR and Western blot showed that serum containing Fuzheng Jiedu decoction could down-regulate the protein expression of BCL-2. Fuzheng Jiedu decoction could reduce the protein expression of P-gp on the K562/A02 cell membrane. CONCLUSION: serum containing Fuzheng Jiedu decoction can promote K562/A02 cell apoptosis, its mechanism of inducing apoptosis may be related with the inhibition of BCL-2 and P-gp protein expression.

[Effects of electroacupuncture on the immunoreactivity of focal cutaneous CB2 receptor positive cells in arthritis rats].

OBJECTIVE: To study the effects of electroacupuncture (EA) on the immunoreactivity of focal cutaneous cannabinoid receptor 2 (CB2) positive cells in adjuvant arthritis (AA) rats. METHODS: A total of 48 adult female SD rats were randomly divided into control group (n = 12), model group (n = 12), acupoint group (n = 12), and non-acupoint group (n = 12). Arthritis model was established by hypodermic injection of complete Freund's adjuvant (CFA, 50 microL) into the left ankle joint. EA (2/ 15 Hz, 1 mA) was applied to "Huantiao" (GB 30) and "Yanglingquan" (GB 34) or two control points 5 mm left to GB30 and GB34 on the diseased side for 30 min, once every other day from the second day on after injection of CFA. Behavioral performance (pain test score) was assesed by using dorsiflexion and plantarflexion pain tests. On the 6th and 16th day after injection of CAF, the animals were anesthetized with 20% urethane (1 g/kg) for collecting the focal skin and subcutaneous tissue samples which were cut into sections (5 microm) to be stained with HE (haematoxylin & cosin) method and immunohistochemical technique respectively for observing changes of the focal cells of the inflamatory tissue and the immunoactivity of the focal cutaneous CB receptor positive cells. RESULTS: 1) In comparison with control group, the scores of both dorsiflexion and plantarflexion pain tests in model, acupoint and non-acupoint groups increased evidently after modeling (P < 0.05). Compared with model group, the scores of dorsiflexion test on the 3rd day and 5th day and plantarflexion test on the 5th day in EA-acupoint group were considerably lower (P < 0.05), and the scores of the two tests on the 3rd day and 5th day of acupoint group were also markedly lower than those of non-acupoint group (P < 0.05), suggesting a marked pain-relief after EA; while no significant differences were found between non-acupoint and model groups. 2) HE staining showed that the inflammatory cells in the dermal layer of the focus of acupoint group were evidently fewer than those of model and non-acupoint groups on the 6th day and 16th day after modeling. 3) The immunohistochemical results revealed that compared with control group, on the 6th day, the percentages of CB2 receptor positive cell area in the focus of model, non-acupoint and acupoint groups were significantly higher (P < 0.05), and that of acupoint group was markedly higher than those of model and non-acupoint groups (P < 0.05), suggesting further upregulation of the expression of CB2 receptor positive cells; no significant differences of percentages of CB2 receptor immunoreaction positive cell area among model, non-acupoint and acupoint groups were found on the 16th day (P > 0.05). CONCLUSION: EA of GB30 and GB34 can raise the immunoactivity of cutaneous CB2 receptor positive cells in the inflammatory tissue which maybe contribute to its effects in relieving inflammatory pain and suppressing focal inflammation and adjusting the balance of nociception and antinociception in AA rats.