[Location, selection, and comparative anatomy of "Taixi" (KI3), "Shuiquan" (KI5), "Fuliu" (KI7), "Jiaoxin" (KI8), "Zhubin" (KI9), and "Yingu"(KI10) acupoints in rabbits].

OBJECTIVE: To investigate the. METHODS: for locating and selecting the acupoints of "Taixi" (KI3), "Shuiquan" (KI5), "Fuliu" (KI7), "Jiaoxin" (KI8), "Zhubin" (KI9), and "Yingu" (KI10) and the morphological structure of these acupoints in rabbits. MethodsAccording to the WHO and national standards for human acupoints and rabbit X-ray images, acupoint locations were marked using the anatomical landmarks on body surface in 10 New Zealand rabbits. The acupoints were dissected to compare the homologous and analogous tissue between rabbits and human body and thus correct the locations of these acupoints. Potentials were measured for the 10 New Zealand rabbits at the corrected locations of the acupoints and around the acupoints, and the final locations of these acupoints were determined by comparing the anatomical results and the data of potentials. Anatomical observation was performed after marking, and the relationship between acupuncture needle and adjacent structure was observed. RESULTS: "Taixi" was located in the ankle area, at the midpoint between the prominence of the medial malleolus and the calca-neal tendon; "Shuiquan" was located in the calcaneal area below "Taixi" in the depression anterior to the calcaneal tuberosity; "Fuliu" was located at the medial side of the calf, at 2 cun above the prominence of the medial malleolus anterior to the calcaneal tendon; "Jiaoxin" was located at the medial side of the calf, at 2 cun above the prominence of the medial malleolus and in the depression posterior to the medial border of the tibia; "Zhubin" was located at the medial side of the calf, at 5 cun above the medial malleolus on the line between "Taixi" and "Yingu"; "Yingu" was located at the medial side of the knee, at the posterior-inferior border of the semitendinosus tendon on the popliteal crease. The results of skin potentials at the acupoints suggested that "Taixi", "Shuiquan", "Fuliu", and "Zhubin" were high-reliability acupoints, "Jiaoxin" was a medium-reliability acupoint, and "Yingu" was a low-reliability acupoint. CONCLUSION: Comparative anatomy combined with imaging, surface anatomy, and electrophysiological techniques of acupoints can help with the accurate localization and selection of acupoints in experimental animals, improve the reliability of acupoint location, and enrich the comparative anatomical data of acupoints.

[Hierarchical structure of Five-shu points and Source-point of Triple Energizer Meridian in rabbits].

OBJECTIVE: To study the position and hierarchical structure of Five-shu points, i．e., "Guanchong" (TE1), "Yemen" (TE2), "Zhongzhu" (TE3), "Yangchi" (TE4) and "Zhigou" (TE6), and the Source-point "Tianjing" (TE10) of the Triple Energizer (TE) Meridian in the rabbit. METHODS: Based on WHO Standard Acupuncture Point Locations in the Western Pacific Region (WHO Standard) and National Standard (GB/T 22103-2008) for Acupuncture Point Locations in human body, and combined with X-ray images, the hierarchical structure of Five-shu points and Source-point of the TE Meridian were observed in ten New Zealand rabbits. The acupoint locations were determined by comparing the same name tissues of the rabbits and human body after dissecting the above-mentioned acupoints. After inserting acupuncture needles into the aforementioned acupoints, the relationship between the acupuncture needle and adjacent structure were dissected and measured. RESULTS: "Guanchong" (TE1) was located on the lateral side of the 4th terminal phalanx, and behind the corner of the onyx root. When needled, the penetrated tissues of the acupuncture needle are skin, superficial fascia, deep fascia, and the root region of the 4th phalanx, respectively. "Yemen" (TE2) was located between the 4th and 5th onyxes, at the depression of intersection of coat hair superior to the fingerweb edge. When needled, the penetrated tissues of the acupuncture needle are the superficial fascia, deep fascia and lumbrical muscles of the forepaw, respectively. "Zhongzhu" (TE3) was located between the 4th and 5th metacarpal bones, at the depression proximal to the 4th metacarpophalangeal joint. When needled, the penetrated tissues are skin, superficial fascia, deep fascia, and lumbrical muscle, respectively. "Yangchi" (TE4) was positioned at the dorsal side of the forepaw, and the surface connection line between the accessory and radial bones intersected with the depression of the extensor digitorum communis on the ulnar side. When needled, the penetrated tissues are skin, superficial fascia, deep fascia, and the ulnar side of the common extensor tendon, respectively. "Zhigou" (TE6) was positioned between the radius and ulna, on the posterior aspect of the forelimb and 3 Bone-cun proximal to the distal dorsal forepaw crease. When needled, the penetrated tissues are skin, superficial fascia, deep fascia, and the extensor digitorum, respectively. "Tianjing" (TE10) was located at the junction of the body of humerus and the lateral condyle of humerus, on the posterior aspect of the elbow and proximal to the prominence of the olecranon. When needled, the penetrated tissues are skin, superficial fascia, deep fascia, and triceps brachii muscle, respectively. CONCLUSION: The Five-shu points and Source-point of the TE Meridian on the forelimb in rabbits are innervated by the cutaneous branches of the ulnar radial nerve and the medial brachial cutaneous nerve at the superficial layer, and by the branches of the ulnar nerve and radial nerve in the deep layer, accompanied with cephalic vein and forearm blood vessels and their branches.

Endothelial-Dependent and Independent Vascular Relaxation Effect of Tetrahydropalmatine on Rat Aorta.

Tetrahydropalmatine (THP) is an active natural alkaloid isolated from Corydalis yanhusuo W.T. Wang which has been widely used for treating pain and cardiovascular disease in traditional Chinese medicine. Previous studies suggested THP have various pharmacological effects in neural and cardio tissue while the vascular reactivity of THP was not fully established. The present study found that THP relaxed rat aorta which contracted by phenylephrine (Phe), KCl, and U46619. The vascular relaxation effect of THP was partially attenuated by PI3K inhibitor wortmannin, Akt inhibitor IV, endothelial nitric oxide synthetase (eNOS) inhibitor L-NAME, guanylate cyclase inhibitors and the mechanical removal of endothelium. Also, the eNOS substrate L-arginine reversed the inhibition effect of L-NAME on THP-induced vascular relaxation. THP also induced intracellular NO production in human umbilical vein endothelial cells. However, Pre-incubation with ß-adrenergic receptor blocker propranolol, angiotensin II receptor 1 (AT1) inhibitor losartan, angiotensin II receptor 2 (AT1) inhibitor PD123319 or angiotensin converting enzyme inhibitor enalapril enhanced the vascular relaxation effect of THP. THP did not affect the angiotensin II induced vascular contraction. Cyclooxygenase-2 (COX2) inhibitor indomethacin did not affect the vascular relaxation effect of THP. Furthermore, pre-treatment THP attenuated KCl and Phe induced rat aorta contraction in standard Krebs solution. In Ca2+ free Krebs solution, THP inhibited the Ca2+ induced vascular contraction under KCl or Phe stress and reduced KCl stressed Ca2+ influx in rat vascular smooth muscle cells. THP also inhibited intracellular Ca2+ release induced vascular contraction by blocking Ryr or IP3 receptors. In addition, the voltage-dependent K+ channel (Kv) blocker 4-aminopyridine, ATP-sensitive K+ channel (KATP) blocker glibenclamide and inward rectifying K+ channel blocker BaCl2 attenuated THP induced vascular relaxation regardless of the Ca2+-activated K+ channel (KCa) blocker tetraethylammonium. Thus, we could conclude that THP relaxed rat aorta in an endothelium-dependent and independent manner. The underlying mechanism of THP relaxing rat aorta involved PI3K/Akt/eNOS/NO/cGMP signaling path-way, Ca2+ channels and K+ channels rather than COX2, ß-adrenergic receptor and renin-angiotensin system (RAS). These findings indicated that THP might be a potent treatment of diseases with vascular dysfunction like hypertension.

Schisantherin A protects against 6-OHDA-induced dopaminergic neuron damage in zebrafish and cytotoxicity in SH-SY5Y cells through the ROS/NO and AKT/GSK3ß pathways.

ETHNOPHARMACOLOGICAL RELEVANCE: The fruit of Schisandra chinensis (Turcz.) Baill, has been traditionally used in management of liver diseases and ageing associated neurodegeneration. The bioactive compound from this medicinal plant would be valuable for its potential use in prevention and treatment of Parkinson׳s disease. AIM OF THE STUDY: The overall objective of the present study was to understand the neuroprotective effect of schisantherin A, a dibenzocyclooctadiene lignan from the fruit of S. chinensis (Turcz.) Baill, and to elucidate its underlying mechanism of action. MATERIAL AND METHODS: This study investigated the protective effect of schisantherin A against selective dopaminergic neurotoxin 6-hydroxydopamine (6-OHDA)-induced neural damage in human neuroblastoma SH-SY5Y cells and zebrafish models. Oxidative stress and related signaling pathways underlying the neuroprotective effect were determined by multiple biochemical assays and Western blot. RESULTS: Pretreatment with schisantherin A offered neuroprotection against 6-OHDA-induced SH-SY5Y cytotoxicity. Moreover, schisantherin A could prevent 6-OHDA-stimulated dopaminergic neuron loss in zebrafish. Our mechanistic study showed that schisantherin A can regulate intracellular ROS accumulation, and inhibit NO overproduction by down-regulating the over-expression of iNOS in 6-OHDA treated SH-SY5Y cells. Schisantherin A also protects against 6-OHDA-mediated activation of MAPKs, PI3K/Akt and GSK3ß. CONCLUSION: These findings demonstrate that schisantherin A may have potential therapeutic value for neurodegenerative diseases associated with abnormal oxidative stress such as Parkinson׳s disease.

Danshensu protects against 6-hydroxydopamine-induced damage of PC12 cells in vitro and dopaminergic neurons in zebrafish.

The overproduction of reactive oxygen species (ROS) has been implicated in the development of neurodegenerative diseases such as Parkinson's disease (PD) and Alzheimer's disease (AD). Previous studies have indicated that danshensu (beta-3,4-dihydroxyphenyl-lactic acid), a main hydrophilic component of the Chinese materia medica Radix Salviae Miltiorrhizae (Danshen, Pharmacopoeia of PR China), has ROS scavenging and antioxidant activities, however its mechanism of action was not clear. In this study, we investigated whether the protective effects of danshensu against neurotoxin 6-hydroxydopamine (6-OHDA)-induced oxidative stress involved the Nrf2/HO-1 pathways. Pretreatment with danshensu in PC12 cells significantly attenuated 6-OHDA-induced cytotoxicity and the production of ROS. Danshensu activated the nuclear translocation of Nrf2 to increase heme oxygenase-1 (HO-1), conferring protection against ROS. Danshensu induced the phosphorylation of Akt, and its cytoprotective effect was abolished by PI3K, Akt and HO-1 inhibitors. These results confirmed the crucial role of PI3K/Akt and HO-1 signaling pathways as the underlying mechanistic action of danshensu. Taken together, the results suggest that danshensu enhances HO-1 expression to suppress 6-OHDA-induced oxidative damage via PI3K/Akt/Nrf2 signaling pathways. Moreover, 6-OHDA-induced dopaminergic neuronal loss in zebrafish could be reduced by danshensu, further supporting the neuroprotective potential of danshensu.

[Use of zebrafish models for the research of traditional Chinese medicine].

Traditional Chinese medicine (TCM) exhibits a broad range of effects on biological activity that is probably due to interactions of complex chemical constituents with multiple targets in the body. Understanding the active chemical constituents in TCM is very important in providing rationales for the clinical usage of TCM. The zebrafish (Danio rerio) has recently become a popular model in the field of drug screening, specifically emerging as an important vertebrate model for in vivo high-content drug screening of multiple efficacy parameters and whole-organism toxicity. The authors also discussed the advantages of the zebrafish model for evaluating drug metabolism. Zebrafish usage in TCM screening should be a viable approach that helps identify active chemical markers, biological pathways and mechanistic actions of TCM.

[Macroscopical anatomic structure of acupoint "Shangyang" (LI 1), "Erjian" (LI 2), "Sanjian" (LI 3), "Hegu" (LI 4), and "Quchi" (LI 11) regions in the forelimb in rabbits].

OBJECTIVE: To observe the structure of acupoints: "Shangyang" (LI 1), "Erjian" (LI 2), "Sanjian" (LI 3), "Hegu" (LI 4) and "Ouchi" (LI 11) of the Yangming Meridian in the rabbit's forelimb. METHODS: The acupoints were first located in accordance with the related atlas of rabbits and confirmed by low resistance and higher electric potential determinations by using a "multipurpose electronic acupoint detection and therapeutic instrument". Then the animals under anesthesia were killed by intravenous injection of air embolism, perfused with 5% natrium citricum first, acetic ether containing acrylonitrile-butadiene-styrene copolymer, and 5% dicapryl phthalate, blue and red pigments for paintings via subclavian artery, respectively. After routinely inserting an acupuncture needle into the acupoint, the local tissues (muscles, blood vessels and nerves) of the acupoints mentioned above were dissected layer by layer and their relations with the needle were observed under microscope. RESULTS: "Erjian" (LI 2) is situated at the depression site distal to the second metacarpophalangeal joint on the radial side. "Sanjian" (LI 3) is located at the depression site proximal to the second metacarpophalangeal joint on the radial side. The shallow-layers of "Shangyang" (LI 1), "Erjian" (LI 2), "Sanjian" (LI 3), "Hegu" (LI 4) and "Quchi" (LI 11) mainly contain cephalic vein and shallow branches of the radial nerve, and their deep layers chiefly contain radial artery and its branches, and the median nerve. CONCLUSION: "Shangyang" (LI 1), "Erjian" (LI 2), "Sanjian" (LI 3), "Hegu" (LI 4) and "Quchi" (LI 11) have a close association with the cephalic vein,radial artery,radial vein and its branches,superficial branch of radial nerve and the median nerve,which constitutes the morphological basis of the five points.