This study investigated the therapeutic effect of Shegan Mahuang Decoction(SGMHD) on cold-induced asthma in rats and explored its underlying mechanism. Seventy-two healthy male SD rats of specific pathogen free(SPF) grade were randomly divided into a blank group, a model group, a positive control group(dexamethasone, 0.4 mg·kg~(-1)), and low-, medium-, and high-dose SGMHD groups(3.2, 6.4, and 12.8 g·kg~(-1)). The blank group received saline, while the other groups were sensitized by intraperitoneal injection of ovalbumin(OVA) solution. Subsequently, the rats were placed in a cold chamber adjustable to 0-2 ℃, and OVA solution was ultrasonically nebulized to induce cold-induced asthma in rats. After three weeks of treatment, the general behaviors of rats were observed. Hematoxylin-eosin(HE) staining was used to evaluate pathological changes in lung tissues, periodic acid-Schiff(PAS) staining assessed mucin changes, and Masson staining was performed to examine collagen deposition. Enzyme-linked immunosorbent assay(ELISA) was used to measure the levels of the inflammatory factors interleukin-4(IL-4) and vascular endothelial growth factor(VEGF) in serum and bronchoalveolar lavage fluid(BALF). Real-time quantitative polymerase chain reaction(RT-PCR) was employed to assess the mRNA expression levels of transient receptor potential vanilloid subfamily member 1(TRPV1), nuclear respiratory factor 1(NRF-1), and mitochondrial transcription factor A(mtTFA) in lung tissues. Western blot was used to measure the protein expression levels of TRPV1, NRF-1, and mtTFA in lung tissues. Compared with the blank group, the model group exhibited signs of rapid respiration, increased frequency of defecation with looser stools, and disheveled and dull fur. Pathological results showed significant infiltration of inflammatory cells in lung tissues, narrowing of bronchial lumens, increased mucin secretion, and enhanced collagen deposition in the model group. Additionally, the levels of IL-4 and VEGF in serum and BALF were significantly elevated, and the mRNA and protein expression levels of TRPV1, NRF-1, and mtTFA in lung tissues were significantly increased. Compared with the model group, SGMHD improved the behaviors of rats, alleviated pathological changes in lung tissues, mucin production, and collagen deposition, significantly decreased the levels of IL-4 and VEGF in serum and BALF, and reduced the mRNA expression levels of TRPV1, NRF-1, and mtTFA in lung tissues, with the medium-dose SGMHD group showing the most significant effect. Moreover, the protein expression levels of TRPV1, NRF-1, and mtTFA in lung tissues were also reduced, with the medium-dose SGMHD group exhibiting the most significant effect. In conclusion, this study demonstrates that SGMHD can alleviate airway inflammation and inhibit airway remodeling in cold-induced asthma rats. These effects may be associated with the modulation of the TRPV1/NRF-1/mtTFA signaling pathway.
Rats ; Male ; Animals ; Mice ; Interleukin-4/metabolism* ; Vascular Endothelial Growth Factor A/metabolism* ; Rats, Sprague-Dawley ; Asthma/genetics* ; Lung ; Bronchoalveolar Lavage Fluid ; RNA, Messenger/metabolism* ; Collagen/metabolism* ; Mucins/therapeutic use* ; Ovalbumin ; Disease Models, Animal ; Mice, Inbred BALB C ; TRPV Cation Channels/metabolism* ; Drugs, Chinese Herbal

This study aims to explore the pharmacodynamic effect of baicalin on rat brain edema induced by cerebral ischemia reperfusion injury and discuss the mechanism from the perspective of inhibiting astrocyte swelling, which is expected to serve as a refe-rence for the treatment of cerebral ischemia with Chinese medicine. To be specific, middle cerebral artery occlusion(suture method) was used to induce cerebral ischemia in rats. Rats were randomized into normal group, model group, high-dose baicalin(20 mg·kg~(-1)) group, and low-dose baicalin(10 mg·kg~(-1)) group. The neurobehavior, brain index, brain water content, and cerebral infarction area of rats were measured 6 h and 24 h after cerebral ischemia. Brain slices were stained with hematoxylin and eosin(HE) for the observation of pathological morphology of cerebral cortex after baicalin treatment. Enzyme-linked immunosorbent assay(ELISA) was employed to determine the content of total L-glutathione(GSH) and glutamic acid(Glu) in brain tissue, Western blot to measure the content of glial fibrillary acidic protein(GFAP), aquaporin-4(AQP4), and transient receptor potential vanilloid type 4(TRPV4), and immunohistochemical staining to observe the expression of GFAP. The low-dose baicalin was used for exploring the mechanism. The experimental results showed that the neurobehavioral scores(6 h and 24 h of cerebral ischemia), brain water content, and cerebral infarction area of the model group were increased, and both high-dose and low-dose baicalin can lower the above three indexes. The content of GSH dropped but the content of Glu raised in brain tissue of rats in the model group. Low-dose baicalin can elevate the content of GSH and lower the content of Glu. According to the immunohistochemical staining result, the model group demonstrated the increase in GFAP expression, and swelling and proliferation of astrocytes, and the low-dose baicalin can significantly improve this situation. The results of Western blot showed that the expression of GFAP, TRPV4, and AQP4 in the cerebral cortex of the model group increased, and the low-dose baicalin reduce their expression. The cerebral cortex of rats in the model group was severely damaged, and the low-dose baicalin can significantly alleviate the damage. The above results indicate that baicalin can effectively relieve the brain edema caused by cerebral ischemia reperfusion injury in rats, possibly by suppressing astrocyte swelling and TRPV4 and AQP4.
Animals ; Aquaporin 4/genetics* ; Astrocytes ; Brain Edema/drug therapy* ; Brain Ischemia/metabolism* ; Flavonoids ; Infarction, Middle Cerebral Artery/drug therapy* ; Rats ; Rats, Sprague-Dawley ; Reperfusion ; TRPV Cation Channels/therapeutic use*

OBJECTIVE: To observe the effect of electroacupuncture (EA) at "Tianshu" (ST 25) and "Shangjuxu" (ST 37) on mental state, visceral sensitivity and protein expression of nerve growth factor (NGF), tyrosine kinase receptor A (TrkA) and transient receptor potential vanilloid 1 (TRPV1) of colonic tissue in diarrhea-predominant irritable bowel syndrome (IBS-D) rats, and to explore its possible mechanism on treating IBS-D. METHODS: A total of 36 male SD rats of SPF grade were randomized into a blank group, a model group, an EA group and a western medication group, 9 rats in each group. In the model group, the EA group and the western medication group, IBS-D model was established by enema of dinitrobenzene sulfonic acid (DNBS) combined with chronic restraint stress method. In the EA group, EA was applied at "Tianshu" (ST 25) and "Shangjuxu" (ST 37), with disperse-dense wave, in frequency of 2 Hz/100 Hz, 20 min each time, once a day for 7 days. In the western medication group, pinaverium bromide suspension was given by gavage (15 mg•kg^-1•d^-1) for 7 days. Before and after model establishment, and after intervention, the body mass, 24 h food intake and fecal water content were observed, the visceral sensitivity was detected by abdominal withdrawal reflex (AWR); after intervention, the mental state was evaluated by elevated plus maze (EPM) test, the protein expression of NGF, TrkA and TRPV1 was detected by immunohistochemistry and Western blot in the 4 groups. RESULTS: After model establishment, compared with the blank group, the body mass and 24 h food intake were decreased (P<0.05), first systolic latency of AWR was shortened and number of contraction wave of AWR was increased (P<0.05), and fecal water content was increased (P<0.05) in the model group, the EA group and the western medication group. After intervention, compared with the blank group, open arm residence time ratio (OT%) of EPM was decreased (P<0.05) and protein expression of NGF, TrkA, TRPV1 in colonic tissue was increased in the model group (P<0.05); compared with the model group, the body mass and 24 h food intake were increased (P<0.05), first systolic latency of AWR was lengthened and number of contraction wave of AWR was decreased (P<0.05), the fecal water content was decreased (P<0.05), OT% of EPM was increased (P<0.05), and protein expression of NGF, TrkA, TRPV1 in colonic tissue was decreased (P<0.05) in the EA group and the western medication group. CONCLUSION Electroacupuncture at "Tianshu" (ST 25) and "Shangjuxu" (ST 37) can relieve the anxiety and depression-like behaviors in IBS-D rats, down-regulate the protein expression of NGF, TrkA, TRPV1 in colonic tissue, so as to reduce the visceral sensitivity and relieve symptoms.
Male ; Rats ; Animals ; Receptor Protein-Tyrosine Kinases ; Rats, Sprague-Dawley ; Irritable Bowel Syndrome/therapy* ; Sulfonic Acids ; Nerve Growth Factors ; TRPV Cation Channels/genetics*

OBJECTIVE: To analyze the clinical and genetic characteristics of a patient featuring autosomal dominant Olmsted syndrome. METHODS: Clinical features of the patient was reviewed. High-throughput sequencing was carried out to detect potential genetic variants. RESULTS: The proband, a 12-year-old girl, featured excessive keratinization on hands and feet, contracture of finger joints, and abnormal position and residual contraction of the fifth toes. Skin biopsy showed significant hyperkeratosis, epidermal hyperplasia, and mild interepidermal cell edema. A de novo heterozygous missense variant c.2016G>T(p.Met672Ile) was identified in the TRPV3 gene by high-throughout sequencing. The result was verified by Sanger sequencing. CONCLUSION The destructive palmoplantar keratosis in the child may be attributed to the c.2016G>T(p.Met672Ile) variant of the TRPV3 gene. Aboving finding has provided new evidence for the correlation of genetic variants with clinical phenotypes of Olmsted syndrome.
Child ; Female ; Heterozygote ; Humans ; Keratoderma, Palmoplantar/genetics* ; Skin ; Syndrome ; TRPV Cation Channels/genetics*

The purpose of the present study was to investigate the effect of transient receptor potential vanilloid 4 (TRPV4) channel on the permeability of pulmonary microvascular endothelial cells (PMVECs) in rats with chronic hypoxia-induced pulmonary hypertension (CHPH), so as to clarify the mechanism of vascular endothelial dysfunction during the occurrence of pulmonary hypertension (PH). CHPH rat model was established by exposure to chronic hypoxia (CH) for 21 days. Primary PMVECs were cultured by adherent tissue blocks at the edge of the lung. The permeability coefficient of primary cultured PMVECs was detected by fluorescein isothiocyanate (FITC)-dextran. The structure of tight junction (TJ) was observed by transmission electron microscope. The expression of TRPV4 and TJ-related proteins, such as, Occludin, Claudin-5, ZO-1 were examined by real-time fluorescence quantitative PCR and Western blotting. The intracellular calcium concentration ([Ca
Animals ; Endothelial Cells ; Hypertension, Pulmonary ; Hypoxia/complications* ; Lung ; Permeability ; Rats ; TRPV Cation Channels/genetics*

To explore the role of vascular endothelial growth factor (VEGF) in diabetic peripheral neuropathic pain in rats.
 Methods: Twenty-four adult male Sprague-Dawley rats aged 8 weeks were randomly divided into 3 groups (n=8 per group). The control group (C group): rats were intraperitoneally injected with sodium citrate solution at 10 mL/kg; the model group (M group): rats were intraperitoneally injected with streptozotocin at 65 mg/kg; the treatment group (T group): rats received intraperitoneal injection of anti-VEGF antibody (10 mg/kg) at the 1st, 3rd, 7th, 10th day after STZ treatment. Meanwhile, rats of C and M group were received with the same volume of sodium citrate solution. Blood glucose was measured before 1 day or at the 1st, 3rd, 7th or 14th day after receiving STZ. Body weight, paw withdrawal mechanical threshold (PWMT) and paw withdrawal thermal latency (PWTL) were measured before 1 day or at the 1st, 3rd, 5th, 7th, 10th or 14th day after receiving STZ. All lumbar spinal cords were dissected to examine the p-protein kinase B (p-Akt) and transient receptor potential vanilloid 1 (TRPV1) expression by Western blot.
 Results: After injection with STZ, the body weight showed significant differences at some time point between the M, T or C group (P<0.01); body weight of rat in the C group were increased gradually. Compared with the C group, the fast blood glucose in the M or the T Group at the same time points were increased significantly (P<0.01). The PWMT and PWTL of the M, T or C group were significant difference among various time points (P<0.01). The PWMT and PWTL in the M or T group were obviously reduced compared with those in the C group (P<0.01). Compared with the M group, the PWMT and PWTL in the T group were increased at the 10th or 14th day (P<0.01 or P<0.05). Compared with the C group, the p-Akt and TRPV1 levels in the M and T group were increased (P<0.01). Compared with the M group, p-Akt and TRPV1 levels in T group were decreased (P<0.01).
 Conclusion: VEGF is able to regulate the expression of TRPV1 through PI3K/Akt pathway, which contributes to diabetic peripheral neuropathic pain in rats. Anti-VEGF treatment may be useful for alleviation of diabetic peripheral neuropathic pain.
Animals ; Antibodies ; pharmacology ; therapeutic use ; Diabetes Mellitus, Experimental ; chemically induced ; Diabetic Neuropathies ; chemically induced ; drug therapy ; Gene Expression Regulation ; drug effects ; Male ; Phosphatidylinositol 3-Kinases ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; TRPV Cation Channels ; genetics ; Vascular Endothelial Growth Factor A ; metabolism

Olmsted syndrome (OS) is a rare congenital skin disorder characterized by severe palmoplantar and periorificial keratoderma, alopecia, onychodystrophy, and severe pruritus. Recently, pathogenic ‘gain-of-function‘ mutations of the transient receptor potential vanilloid 3 gene (TRPV3), which encodes a cation channel involved in keratinocyte differentiation and proliferation, hair growth, inflammation, pain and pruritus, have been identified to cause OS. Due to the rarity, the pattern of inheritance of OS is still unclear. We report a case of OS in a 3-year-old Korean girl and its underlying gene mutation. The patient presented with a disabling, bilateral palmoplantar keratoderma with onychodystrophy. She also exhibited pruritic eczematous skin lesions around her eyes, ears and gluteal fold. Genetic analysis identified a heterozygous p.Gly568Val missense mutation in the exon 13 of TRPV3. To our knowledge, this is the first case of OS in the Korean population showing a missense mutation p.Gly573Ser.
Abnormalities, Multiple/*genetics ; Base Sequence ; Child, Preschool ; Female ; Heterozygote ; Humans ; Keratoderma, Palmoplantar/genetics ; Lipid Droplets/ultrastructure ; Mutation, Missense/*genetics ; Skin/pathology/ultrastructure ; Syndrome ; TRPV Cation Channels/*genetics

Capsaicin in chili peppers bestows the sensation of spiciness. Since the discovery of its receptor, transient receptor potential vanilloid 1 (TRPV1) ion channel, how capsaicin activates this channel has been under extensive investigation using a variety of experimental techniques including mutagenesis, patch-clamp recording, crystallography, cryo-electron microscopy, computational docking and molecular dynamic simulation. A framework of how capsaicin binds and activates TRPV1 has started to merge: capsaicin binds to a pocket formed by the channel's transmembrane segments, where it takes a "tail-up, head-down" configuration. Binding is mediated by both hydrogen bonds and van der Waals interactions. Upon binding, capsaicin stabilizes the open state of TRPV1 by "pull-and-contact" with the S4-S5 linker. Understanding the ligand-host interaction will greatly facilitate pharmaceutical efforts to develop novel analgesics targeting TRPV1.
Binding Sites ; Capsaicin ; chemistry ; pharmacokinetics ; Humans ; Hydrogen Bonding ; Protein Binding ; TRPV Cation Channels ; chemistry ; genetics ; metabolism

OBJECTIVETo study the effects of the change in transient receptor potential vanilloid 1 (TRPV1) channel activity on the degree of airway inflammation in asthmatic mice.

METHODSBALB/c mice were randomly divided into control, asthma, capsaicin (TRPV1 agonist), capsazepine (TRPV1 antagonist), and dexamethasone groups. The asthmatic mouse model was established by intraperitoneal injection of mixed ovalbumin-aluminium hydroxide solution and ultrasonic atomization with OVA for sensitization and challenge. The capsaicin, capsazepine, and dexamethasone groups were given intraperitoneal injection of capsaicin (30 μg/kg), capsazepine (10 μmol/kg), and dexamethasone (2 mg/kg) respectively, at 30 minutes before challenge. Hematoxylin and eosin staining was used to observe the degree of pulmonary inflammation. ELISA was used to measure the content of interleukin-8 (IL-8) and interleukin-13 (IL-13) in bronchoalveolar lavage fluid (BALF). Real-Time PCR was used to measure the relative content of TRPV1 mRNA in lung tissue.

RESULTSCompared with the asthma group, the capsazepine and dexamethasone groups showed reduced pulmonary inflammation, while the capsaicin group showed aggravated pulmonary inflammation. Compared with the control group, the asthma and capsaicin groups showed increases in the content of IL-13 and IL-8 in BALF and the mRNA expression of TRPV1 in lung tissue (P<0.05). Compared with the asthma group, the capsazepine and dexamethasone groups showed reductions in the content of IL-13 and IL-8 in BALF and the mRNA expression of TRPV1 in lung tissue (P<0.05). The capsaicin group showed increases in the content of IL-13 and IL-8 in BALF (P<0.05).

CONCLUSIONSTRPV1 channel agonist and antagonist can influence the degree of airway inflammation in asthmatic mice. Dexamethasone may reduce airway inflammation through regulating TRPV1 level.

Animals ; Asthma ; etiology ; Female ; Interleukin-13 ; analysis ; Interleukin-8 ; analysis ; Lung ; pathology ; Mice ; Mice, Inbred BALB C ; RNA, Messenger ; analysis ; TRPV Cation Channels ; genetics ; physiology

OBJECTIVETo explore the analgesic effect and action mechanism of electroacupuncture (EA) on gastric ulcer rats with liver-depression syndrome.

METHODSThrough open-field experimental method, 45 qualified SPF-grade male SD rats were selected and divided into a blank group, a model group and an EA group according to random number table method, 15 rats in each group. The model of gastric ulcer rats with liver-depression syndrome was established in the model group and the EA group by using chronic unpredictable stimulation combined with acetic acid burning method. Rats in the blank group did not receive intervention. Rats in the model group were treated with fixation and immobilization for 13 days. Rats in the EA group were treated with EA at "Liangqiu" (ST 34) and "Ganshu" (BL 18); EA voltage was 2 V; disperse-dense wave was selected with 4 Hz of disperse wave and 15 Hz of dense wave, and the intensity of EA was according to the slight vibration of local skin and; muscles; the needles were retained for 20 min, once a day for consecutive 6 days; there was an interval of 1 day' and the treatment was given for 2 weeks. The general condition, open-field experimental result and gastric ulcer index were observed; the western blotting method was applied to measure the expression of vanilloid receptor subtype 1 (VR1) in hypothalamus and gastric antral mucosal, and ELISA method was applied to test the expression of 5-hydroxytryptamine (5-HT) and norepinephrine (NE) in hippocampus.

RESULTSAfter model establishment, the general behavior condition in the model group was inferior to that in the blank group, which was obviously improved after EA. The range of motion in the model group was less than that in the blank group (P<0.01) while that in the EA group was higher than that in the model group (P<0.01). The ulcer inhibition rate was. 54.95%, and the ulcer index in the EA group was lower than that in the model group (P<0.01). Compared with; the blank group, the expression of VR1 in hypothalamus and gastric antral mucosal in the model group was increased (P<0.05); compared with the model group, the expression of VR1 in the EA group was reduced (P<0.05). Compared with the blank group, the expression of 5-HT an NE in hippocampus in the model group was significantly reduced (both P<0.01); compared with the model group, the expression of 5-HT and NE in the EA group was increased (both P<0.01).

CONCLUSIONEA at "Liangqiu" (ST 34) and "Ganshu" (BL 18) has certain analgesic effect in gastric ulcer rats with liver-depression syndrome, which is likely to be related with lowering the contents of VR1 in hypothalamus and gastric antral mucosal and increasing the content of 5-HT and NE in hippocampus.

Acupuncture Analgesia ; Acupuncture Points ; Animals ; Electroacupuncture ; Humans ; Liver ; physiopathology ; Male ; Nociceptive Pain ; genetics ; metabolism ; physiopathology ; therapy ; Rats ; Rats, Sprague-Dawley ; Serotonin ; metabolism ; Stomach Ulcer ; genetics ; metabolism ; physiopathology ; therapy ; TRPV Cation Channels ; genetics ; metabolism