Herbal complex 'Buyang Huanwu Tang' improves motor endplate function of denervated-dependent skeletal muscle atrophy in rat.

Denervated-dependent skeletal muscle atrophy is a disease induced by skeletal muscle associated peripheral neuro-disconnection. Its specific molecular mechanisms remain unknown. The treating for denervated-dependent skeletal muscle atrophy is applied with an herbal complex Buyang Huanwu Tang used in traditional Chinese medicine and subjected to the established denervated-dependent skeletal muscle atrophy in rat models, and the therapeutic effects and associated mechanisms were evaluated in the pathogenesis of denervated-dependent skeletal muscle atrophy. Denervated-dependent skeletal muscle atrophy in rats was established and randomly divided into eight groups, including Normal control, Model, Positive control, Model + Buyang Huanwu Tang, Model + astragalus extracts, Model + Buyang Huanwu Tang-astragalus, Buyang Huanwu Tang + LY294002, and astragalus extract + LY294002 group. Hematoxylin-eosin staining and quantitative RT-PCR (qRT-PCR) assay were used to examine the inflammatory response of muscle tissues. Quantitative RT-PCR and Western blotting assay were utilized to analyze mRNA and protein expression. Immunohistochemistry assay was used to detect molecule expression in anterior cervical muscle tissues. Motor endplate activity was examined using the wholemount acetylcholinesterase staining method. The wet mass ratio of anterior cervical muscle was measured. The results indicated that Buyang Huanwu Tang treatment significantly alleviated inflammatory response, enhanced acetylcholinesterase activity, and motor endplate functions, and promoted wet mass of anterior cervical muscle compared to denervated-dependent skeletal muscle atrophy rat models (P < 0.05). Buyang Huanwu Tang regulated molecules of PI3K/PKB/GSK3ß/FOXO1 signaling pathway. Buyang Huanwu Tang significantly reduced muscle atrophy F-box protein, MuFR-1, Bax and caspase 9 expression, significantly enhanced Bcl-2 expression, and remarkably increased element-binding protein and vascular endothelial growth factor levels, compared to Model group (P < 0.05). Buyang Huanwu Tang suppressed caspase 9 and caspase 3 activity and associated apoptosis. Moreover, PI3K specific blocker, LY294002, significantly inhibited the effects of Buyang Huanwu Tang on the above molecule expression (P < 0.05). In conclusion, Buyang Huanwu Tang improved motor endplate functions of denervated-dependent skeletal muscle atrophy rat model through suppressing mitochondria-mediated apoptosis and activating PI3K/PKB/FOXO1 signaling pathway.

Buyang Huanwu Tang alleviates inflammation and improves motor endplate functions in DSMA rat models by activating several biological molecules and associated signaling pathways.

Denervated-dependent skeletal muscle atrophy (DSMA) is considered to be the neuro-disconnection of skeletal muscle. This study aimed to investigate the protective effects of Buyang Huanwu Tang (BYHWT) on the DSMA and clarify associated molecular and genetic mechanisms. DSMA rat models were established according to the previously published study and divided into Model group and BYHWT group. Meanwhile, normal rats were assigned as Normal control (NC) group. Hematoxylin and eosin (HE) staining was used to examine inflammatory responses. Motor endplate activity was evaluated with wholemount acetylcholinesterase (AChE) staining. Mass-spectrometry analysis was conducted to compare differentially expressed proteins. RNAs were prepared and applied to gene functional analysis. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were employed to analyze biological functions. The results indicated that BYHWT remarkably alleviated inflammatory responses and significantly improved motor endplate function, compared to that in DSMA Model rats (P<0.05). In BYHWT group, there were 393 differentially up-regulated and 576 differentially down-regulated molecules compared to that in Model group. Comparing to Model group, the cellular response to interferon-gamma, integral component of plasma membrane and voltage-gated potassium channel activity genes in BYHWT group were the most biological process (BP), cellular component (CC) and molecular function (MF) differential genes, respectively. Fructose/mannose metabolism and glycerolipid metabolism KEGG signaling pathways illustrated the most significant enrichment of differentially expressed genes. In conclusion, BYHWT alleviated the inflammations and improved the motor endplate function of DSMA rats by activating cellular response to interferon-gamma, integral component of plasma membrane and voltage-gated potassium channel activity genes and associated signaling pathways.

[Effect of Buyang Huanwu decoction on NF-κB/MuRFl expression in rats with denervated tibial muscle atrophy].

We aim to study the effect of Buyang Huanwu decoction (Astragali Radix, Angelicae Sinensis Radix, Paeoniae Radix Alba, Pheretima, Chuanxiong Rhizoma, Carthami Flos and Persicae Semen) on NF-κB/MuRFl signaling pathway in the denervated tibial muscle atrophy of rats and explore its protection mechanisms. Sixty Sprague-Dawley rats were subjected to common peroneal nerve crush models of 5 mm injury, and then were randomly divided into six groups for daily intragastric administration of drugs： sham operation group, model group, Buyang Huanwu decoction high, middle and low dose groups, and mecobalamin group (as a positive control). After drug administration for 10 days and 21 days, the gene and protein expression levels of NF-κB and MuRFl in the tibial muscle of rats were detected by qRT-PCR and Western blot respectively. The results showed that, as compared with the model group, RNA and protein expression levels of NF-κB and MuRF1 were significantly reduced in Buyang Huanwu decoction middle and high-dose groups (P<0.05 or P<0.01). These results indicated that Buyang Huanwu decoction could inhibit ubiquitin proteasome system probably by activating NF-κB/MuRF1 signal pathway to protect the denervated tibial muscle atrophy of rats.

[Study on preventive and therapeutic effects of astragali radix on denervated tibial muscle atrophy in rats].

OBJECTIVE: To study the effect of Astragali Radix on the denervated tibial muscle atrophy in rats, and discuss its mechanism. METHOD: Totally 60 SPF-grade Sprague-Dawley rats were selected in the common peroneal nerve crush model, and then randomly divided into 6 groups: Astragali Radix high-dose, medium-dose, low-dose groups, the Mecobalamin group, the model group, and the sham operation group. They were administered with drugs after the operation. At 18 d, the pathological section staining and morphological analysis were performed. The wet-weight ratio and section area of tibial muscles were also measured. The real-time fluorescence quantification was adopted to detect the differential expression between Angptl4 and PI3K genes. RESULT: (1) Wet-weight ratio: The wet-weight ratio in Astragali Radix high-dose, medium-dose groups was much higher than that in the model group (P < 0.05 or P < 0.01). (2) Section area: The sham operation group was higher, with regular morphology; Whereas the model group showed significant decrease, with chaotic structure and obvious connective tissue proliferation; Astragali Radix groups and the mecobalamin group showed relatively small section areas, with chaotic structure and unobvious connective tissue proliferation. Compared with the model group, Astragali Radix groups showed significant increase (P < 0.01). (3) Motor end plate: The sham operation group was in uniform brownish black color and oval or round shape; Astragali Radix medium-dose and high-dose group and the mecobalamin group showed rough line edges; Astragali Radix medium-dose and low-dose groups and the model group showed decline in the number, with irregular morphology, rough line edges and a light color. (4) Angptl4 and PI3K: Compared with the model group, the Astragali Radix high-dose group showed significant increase (P < 0.05). CONCLUSION: Astragali Radix has a significant effect in preventing and treating denervated tibial muscle atrophy. It may delay the muscle atrophy by increasing Angptl4 and PI3K gene expressions.

[Experimental study on Buyang Huanwu decoction ([Chinese characters: see text]) for promoting functional recovery of crushed common peroneal nerve in rats].

OBJECTIVE: To study the effects of Buyang Huanwu Decoction ([Chinese characters: see text]) on promoting functional recovery of crushed common peroneal nerve in rats. METHODS: Thirty Sprague-Dawley rats were subjected to produce common peroneal nerve injuries model,and the length of injury was 5 mm. All the rats were divided into 3 groups: BYHWD group, mecobalamin group and model group. The drugs were given by gavage daily for 18 days. Footprint test was performed at the 18th day after surgery to evaluate toe spread function (TSF). Electrophysiology was performed at the 18th day after operation to determine the nerve conduct velocity (NCV). The wet weight ratio and section area of tibial muscle were also measured. RESULTS: (TSF:At the 18th day after operation, the TSF in BYHWD group (-0.15 +/- 0.07) increased significantly compared with that of model group (-0.25 +/- 0.07) (P < 0.01); the TSF in mecobalamin group (-0.17 +/- 0.08) also increased notably compared with that of model group (P < 0.01).(2) NCV: the NCV in BYHWD group [(18.36 +/- 2.74) m/s] (P < 0.01l) and in mecobalamin group [(16.32 +/- 3.54) m/s] (P < 0.05) also increased significantly compared with that of model group [(9.08 +/- 2.56) m/s]; there was striking variation between model group and mecobalamin group (P < 0.05). (3) Wet weight ratio: the wet weight ratio in BYHWD group [(64.21 +/- 2.92)%] (P < 0.01)and in mecobalamin group [(62.43 +/- 3.21)%] (P < 0.01) all increased significantly compared with that of model group [(54.27 +/- 2.05)%]. (4) The section area of tibial muscle: the section area of tibial muscle in BYHWD group [(654.21 +/- 42.92) cm2] (P < 0.01) and in mecobalamin group [(638.43 +/- 93.21) cm2] (P < 0.01) all increased significantly compared with that of model group [(574.27 +/- 52.05) cm2]; there was also striking variation between model group and mecobalamin group (P < 0.05). CONCLUSION: BYHWD can promotes functional recovery of crushed nerve as a result of accelerating recovery of TSF, raising NCV and delaying the decrease of tibial muscle section area and wet weight ratio.

[Study on the characteristics of population distribution of TCM syndromes and its related factors in patients of depression].

OBJECTIVE: To study the rules of occurrence and development of TCM syndromes in patients of depression by analysing the population distribution and related factors of it. METHODS: A clinical epidemiologic survey was carried out with the materials collected by the four-diagnostic method from patients of depression to gain the syndrome types of 397 patients through dynamic cluster analysis, factor analysis and TCM syndrome differentiation. The constitution of syndrome types in different population and the relation between TCM syndrome types and the clinical types differentiated by Western medicine of depression was analyzed. RESULTS: Six syndrome types were found in 397 patients with depression. They were, ranging in occurrence order, the Gan-Qi stagnation with mental uneasiness type (108 cases, 27.2%); the Xin-Pi deficiency with dampness retention type (94 cases, 23.7%); the Xin-Gan Qi-stagnation with Phlegm retention type (76 cases, 19.1%); the Xin-Gan Qi stagnation with Jing-Luo disharmony type (57 cases, 14.3%); the Xin-Shen Qi stagnation with collateral obstruction type (34 cases, 8.6%); and the Xin-Gan Qi stagnation with Heat interfering mental type (28 cases, 7.1%). There were statistical significance (P < 0.01) in comparison of the syndrome types constitution among patients of different ages or different Western medicinal types (mild, with or without psychotic symptoms), but no significant difference was shown in that among patients of different sexes, with or without familial history of mental diseases, with monophasic or biphasic, initial or recurrent episode (P > 0.05). CONCLUSION: The result obtained in this study is basically in accord with the cognitive rule of TCM theory, it provides a reliable base of syndrome differentiation for treatment.