CN115737627A - Application of alpha-mangostin in preparation of medicine for preventing and treating hypertension and hypertensive nephropathy - Google Patents
Application of alpha-mangostin in preparation of medicine for preventing and treating hypertension and hypertensive nephropathy Download PDFInfo
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Abstract
The invention relates to a new application of alpha-mangostin in preparing a medicament for preventing and treating hypertension and hypertensive nephropathy. The research finds that: alpha-mangostin can inhibit the generation of angiotensin II (ANG II) and up-regulate total nitric oxide synthase (T-NOS), thereby lowering blood pressure and playing a role in preventing and treating hypertension. Animal experiments show that the systolic pressure, diastolic pressure and mean arterial pressure of spontaneous hypertension rats can be obviously reduced by intragastric administration of 1.0mg/kg dose, the blood pressure reducing effect is equivalent to that of 20.0mg/kg losartan, a strong blood pressure reducing effect is shown, the potency of alpha-mangostin is better than that of the clinical blood pressure reducing drug losartan, and the alpha-mangostin is a novel blood pressure reducing drug and can be used for preparing oral blood pressure reducing drugs. In addition, the alpha-mangostin can improve renal tubular injury caused by spontaneous hypertension and has prevention and treatment effects on hypertensive nephropathy. Further cell experiments show that the action mechanism of the alpha-mangostin for improving hypertensive nephropathy is related to the inhibition of renal fibrosis.
Description
Technical Field
The invention relates to application of alpha-mangostin in preparing a medicine for preventing and treating hypertension and hypertensive nephropathy, belonging to the technical field of biological medicines.
Background
Hypertension is a clinical syndrome characterized primarily by elevated systemic arterial blood pressure. At present, the number of people suffering from hypertension is rapidly increased due to various factors such as continuous aging, continuous increase of working pressure, unhealthy life style and the like in China, and the trend of the people getting younger is also realized. By 2017, a survey based on a sample of the population in the 170 ten thousand, 35-75 years (55.6 years on average) showed that 44.7% of patients with hypertension; more seriously, only 7.2% of patients in this population have effective blood pressure control [1] . Hypertension is characterized by high prevalence rate, high disability rate, high death rate, low awareness rate, low treatment rate and low control rate. Hypertension is called as an invisible killer in the medical field, which not only seriously threatens the health of people in China, but also brings huge economic burden to individuals and the country.
The damage of hypertension to target organs is not negligible, the kidney is one of the most vulnerable target organs of hypertension, and the damage of hypertension to the kidney is early shown as damage to renal tubular function [2] . When the renal tubules are damaged, the content of N-acetyl-beta-D-glucosaminidase (NAG) and beta 2-microglobulin (beta 2-MG) markers in urine is increased. The injury of the hypertension to the renal tubules further causes renal fibrosis caused by the tubulointerstitial lesion, and the effective intervention of the renal fibrosis is beneficial to delaying the occurrence of chronic nephropathy and end-stage nephropathy. The tubular Epithelial cell-Mesenchymal Transition (EMT) is a key mechanism for initiating and maintaining the renal interstitial fibrosis, and the occurrence of EMT in the tubular Epithelial cell leads to tubular atrophy and promotes the renal interstitial fibrosis. The renal tubular epithelial marker protein E-cadherin (E-cadherin) is expressed during EMTThe expression of the mesenchymal marker protein alpha-smooth muscle actin (alpha-SMA) and Vimentin is increased, the form of the trans-differentiated renal tubular epithelial cells presents fiber-like change and obtains the migration force. The reversal of EMT can effectively improve renal interstitial fibrosis.
For a long time, researches on pathogenesis and prevention and treatment schemes of hypertensive nephropathy mainly focus on renal blood vessels [3-4] However, recent studies have shown that oxidative stress, inflammation, EMT and further fibrosis of kidney cells (renal tubular epithelial cells, podocytes, endothelial cells, etc.) are also important pathological processes and features in the development and progression of hypertensive nephropathy. The increase of renin-angiotensin system (RAAS) in hypertensive patients leads to abnormal increase of Ang II in vivo, which is the main factor inducing the above-mentioned pathological changes in kidney [5-7] . Activation of TGF-beta signaling pathway is also an important factor for the development of diseases such as fibrosis of kidney, angII has activation effect on TGF-beta and downstream Smad2/3, and AT 1 R inhibitors losartan and ERK 1/2 (MAPK 3/1 ) The inhibitor PD98059 can inhibit the activation of TGF-beta and obviously inhibit the fibrosis tendency of renal tubules [8-9] Thus, the TGF- β signaling pathway is an important downstream mechanism by Ang ii to induce hypertensive nephropathy.
At present, the treatment of hypertension is mainly based on chemical synthesis western medicines, including alpha receptor blocker, beta receptor blocker, diuretic, calcium antagonist, angiotensin converting enzyme inhibitor and angiotensin II receptor antagonist. In clinical practice of treating hypertension by using the above drugs, although most patients can control blood pressure to a certain extent, factors such as combination of multiple drugs, long-term use and drug side effects easily cause poor patient compliance [10] . Furthermore, research studies have shown that approximately 10% of hypertensive patients are drug resistant and that, for such patients, even when treated with three different antihypertensive drugs (including even the maximum tolerated dose of diuretic), their blood pressure still exceeds the normal range [11] . These factors all contribute to the difficulty of continuous control of hypertension.
Modern researches show that part of traditional Chinese medicines or extracts thereof have stable curative effect and few adverse reactions in the aspect of hypertension treatment, and have the characteristics and advantages that western medicines can not be replaced. Therefore, the method has very important significance for searching safe and effective medicines for preventing and treating hypertension and target organ injury caused by hypertension from natural products.
Mangosteen (Garcinia mangostana L.) is a common subtropical fruit, and mangosteen shell has long been used for treating various diseases including abdominal pain, dysentery, diarrhea, suppurations, wound infection and chronic ulcer [12] . The alpha-mangostin is xanthone compound extracted and separated from mangosteen shell, and has rich content up to 121 mg/g of dried pericarp [13] . The chemical structural formula of the alpha-mangostin is as follows:
alpha-mangostin has a wide range of pharmacological activities, including anti-cancer [14] Anti-inflammatory agent [15] And oxidation resistance [16] Anti-diabetic agent [17] And neuroprotective properties in Alzheimer's disease and the like [18-19] The alpha-mangostin induces the apoptosis of colon cancer cells and inhibits the EMT of pancreatic cancer cells by regulating a PI3K/Akt signal channel, thereby inhibiting the migration and invasion of pancreatic cancer cells; by inhibiting Stat3 phosphorylation, cell mitochondrial function damage is induced, and human gastric cancer cell apoptosis is induced; can inhibit the expression of IL-6 and COX-2 in mouse brain tissue caused by LPS, and improve mouse brain inflammation; by inhibiting NF-kB and MAPK signal channels, the symptoms of the ulcerative colitis mouse such as weight loss, diarrhea, bloody stool and the like are effectively improved; can improve the injury of rat cerebral cortex neuron cell axon caused by A beta oligomer, inhibit the generation of ROS in cerebellar granule neuron induced by 3-nitropropionic acid, and protect nerve cell; by reducing isoproterenol-induced oxidative stress, the integrity of the myocardial cell membrane is protected, and the myocardial cell injury is reduced. Through retrieval, the antihypertensive effect of the alpha-mangostin and the improvement of renal injury caused by spontaneous hypertension are not reported.
Reference to the literature
1.Lu J,Lu Y,Wang X,et al.Prevalence,awareness,treatment,and control of hypertension in China:data from 1·7million adults in a population-based screening study(China PEACE Million Persons Project)[J].Lancet(London,England),2017,390(10112):2549-2558.
2. Wanshi, lupolitician, chengxing et al. Relationship of tubulointerstitial injury to glomerular injury in essential hypertension patients [ J ] Western medicine, 2017,29 (3): 351-354,358.
3.Seccia TM,Caroccia B,CalòL.Hypertensive nephropathy.Moving from classic to emerging pathogenetic mechanisms[J].J Hypertens,2017；35(2):205-212.
4.Textor SC,Lerman L.Renovascular hypertension and ischemic nephropathy[J].Am J Hypertens,2010,23(11):1159-1169.
5.Zhuo JL,Ferrao FM,Zheng Y,et al.New frontiers in the intrarenal Renin-Angiotensin system:a critical review of classical and new paradigms[J].Front Endocrinol(Lausanne),2013,4:166.
6.Wang F,Lu X,Liu M,et al.Renal medullary(pro)renin receptor contributes to angiotensin II-induced hypertension in rats via activation of the local renin-angiotensin system[J].BMC medicine,2015,13:278.
7.Hitomi H,Kiyomoto H,Nishiyama A.Angiotensin II and oxidative stress[J].Current opinion in cardiology,2007,22(4):311-315.
8.Yang F,Chung AC,Huang XR,et al.Angiotensin II induces connective tissue growth factor and collagen I expression via transforming growth factor-beta-dependent and-independent Smad pathways:the role of Smad3[J].Hypertension,2009,54(4):877-884.
9.Liu Z,Huang XR,Lan HY.Smad3 mediates ANG II-induced hypertensive kidney disease in mice[J].Am JPhysiol Renal Physiol,2012,302(8):F986-997.
10.Burnier M,Wuerzner G,Struijker-Boudier H,et al.Measuring,analyzing,and managing drug adherence in resistant hypertension[J].Hypertension,2013,62(2):218-225.
11.Persell SD.Prevalence of resistant hypertension in the United States,2003-2008[J].Hypertension,2011,57(6):1076-1080.
12.Pedraza-Chaverri J,Cárdenas-Rodríguez N,Orozco-Ibarra M,et al.Medicinal properties of mangosteen(Garcinia mangostana)[J].Food and chemical toxicology:an international journal published for the British Industrial Biological Research Association,2008,46(10):3227-3239.
13.Ghasemzadenh A,Jaafar HZE,Baghdadi A,et al.Alpha-Mangostin-Rich Extracts from Mangosteen Pericarp:Optimization of Green Extraction Protocol and Evaluation of Biological Activity[J].Molecules(Basel,Switzerland),2018,23(8):1852.
14.Zhang H,Tan YP,Zhao L,et al.Anticancer activity of dietary xanthoneα-mangostin against hepatocellular carcinoma by inhibition of STAT3 signaling via stabilization of SHP1[J].Cell death&disease,2020,11(1):63.
15.Guan H,Li J,Tan X,et al.Natural Xanthoneα-Mangostin Inhibits LPS-Induced Microglial Inflammatory Responses and Memory Impairment by Blocking the TAK1/NF-κB Signaling Pathway[J].Molecular nutrition&food research,2020,64(14):e2000096.
16.Tatiya-Aphiradee N,Chatuphonprasert W,Jarukamjorn K.Ethanolic Garcinia mangostana extract andα-mangostin improve dextran sulfate sodium-induced ulcerative colitis via the suppression of inflammatory and oxidative responses in ICR mice[J].Journal of ethnopharmacology,2021,265:113384.
17.Jiang M,Huang S,Duan W,et al.Alpha-mangostin improves endothelial dysfunction in db/db mice through inhibition of aSMase/ceramide pathway[J].Journal of cellular and molecular medicine,2021,25(7):3601-3609.
18.Wang SN,Li Q,Jing MH,et al.Natural Xanthones from Garcinia mangostana with Multifunctional Activities for the Therapy of Alzheimer's Disease[J].Neurochemical research,2016,41(7):1806-1817.
19.Chen G,Li Y,WangW,et al.Bioactivity and pharmacological properties ofα-mangostin from the mangosteen fruit:a review[J].Expert opinion on therapeutic patents,2018,28(5):415-427.
Disclosure of Invention
Aiming at the technical problems, the invention provides the application of the alpha-mangostin in preparing the medicines for preventing and treating hypertension and/or hypertensive nephropathy.
The technical scheme of the invention is as follows:
firstly, the invention protects the application of the alpha-mangostin in preparing the medicine for preventing and treating hypertension and/or hypertensive nephropathy.
Preferably, the hypertension is spontaneous hypertension and the hypertensive nephropathy is nephropathy caused by spontaneous hypertension.
Alpha-mangostin can inhibit the generation of angiotensin II (ANG II) and up-regulate total nitric oxide synthase (T-NOS), thereby lowering blood pressure and preventing and treating hypertension.
The alpha-mangostin can reduce the content of NAG and beta 2-MG in urine of spontaneously hypertensive rats, thereby having the function of preventing and treating renal tubular injury caused by hypertension.
The alpha-mangostin can increase the expression of Ang II induced E-cadherin of human tubular epithelial cells (HK-2 cells), reduce the expression of alpha-SMA, inhibit the morphological fibrosis of the tubular epithelial cells, inhibit the migration of the tubular epithelial cells and reverse EMT so as to effectively improve the renal interstitial fibrosis caused by hypertension. The alpha-mangostin can inhibit the expression of the angiol-induced TGF-beta receptor of HK-2 cells and the downstream Smad2 thereof, thereby indicating that the mechanism of improving hypertensive renal injury by the alpha-mangostin is related to the inhibition of TGF-beta signaling pathway.
Furthermore, the medicine is a composition which takes alpha-mangostin as an active ingredient and contains auxiliary materials acceptable in the pharmaceutical field, and is any pharmaceutical dosage form.
Experiments prove that: the alpha-mangostin is effective in spontaneous hypertension and renal injury such as renal tubular injury and renal fibrosis caused by the same, and can be administered orally.
Furthermore, the dosage of the alpha-mangostin is 0.4-400 mg/kg.d
The invention has the beneficial effects that:
1) The research of the invention discovers that the compound alpha-mangostin has stronger blood pressure reducing effect, and the blood pressure level of spontaneous hypertension rats can be obviously reduced by intragastric administration of 1.0 mg/kg.
2) The invention applies the alpha-mangostin to the preparation of the antihypertensive drug for the first time, and compared with the existing like products, the potency of the alpha-mangostin is superior to that of the first-line clinical drug losartan. Animal experiments show that the systolic pressure, diastolic pressure and mean arterial pressure of spontaneous hypertension rats can be obviously reduced by intragastric administration of 1.0mg/kg dose, the blood pressure reducing effect is equivalent to that of 20.0mg/kg losartan, a strong blood pressure reducing effect is shown, the potency of alpha-mangostin is better than that of the clinical blood pressure reducing drug losartan, and the alpha-mangostin is a novel blood pressure reducing drug and can be used for preparing oral blood pressure reducing drugs.
3) Meanwhile, alpha-mangostin is also found to improve renal injury caused by hypertension. The research finds that: alpha-mangostin can improve renal tubular injury caused by essential hypertension, and has prevention and treatment effects on hypertensive nephropathy. Further cell experiments show that the action mechanism of the alpha-mangostin for improving hypertensive nephropathy is related to the inhibition of renal fibrosis.
Drawings
FIG. 1 shows the effect of alpha-mangostin on the spontaneous hypertensive rat urine beta 2-MG, NAG and mAB, note: in comparison with the normal group, # P<0.05， ## p is less than 0.01; in comparison to the set of models, * P<0.05， ** P<0.01. the data statistical method is the same as that in Table 1, n =6-8; wherein FIG. 1A is the effect of alpha-mangostin on the urinary beta 2-MG of spontaneously hypertensive rats; FIG. 1B is a graph showing the effect of alpha-mangostin on NAG in urine of spontaneously hypertensive rats; FIG. 1C shows the effect of alpha-mangostin on the mAB of spontaneous hypertensive rat urine
FIG. 2 is a graph showing the effect of alpha-mangostin on the expression levels of E-cadherin and alpha-SMA protein in spontaneously hypertensive rats, note: in comparison with the normal group, # P<0.05， ## p is less than 0.01; compared with model group， * P<0.05， ** P<0.01. The data statistics method is the same as that in Table 1, n =6-8. Wherein FIG. 2A is the effect of alpha-mangostin on the expression level of E-cadherin in spontaneously hypertensive rats; FIG. 2B is a graph showing the effect of alpha-mangostin on the expression level of alpha-SMA protein in spontaneously hypertensive rats;
FIG. 3 is a graph showing the effect of alpha-mangostin on Ang II-induced HK-2 migration, compared to normal, # p is less than 0.05; in comparison to the model set, * P＜0.05， ** P＜0.01，n＝3
FIG. 4 is a graph showing the effect of α -mangostin on the levels of mRNA of EMT marker molecules in HK-2 cells induced by Ang II; as compared with the normal group, the group, ## P＜0.01， ### p is less than 0.001; in comparison to the model set, * P＜0.05， ** P＜0.01， *** p < 0.001,n =3. Wherein FIG. 4A is a graph of the effect of α -mangostin on the levels of E-cadherin mRNA of the cell of HK-2 induced by Ang II; FIG. 4B is a graph of the effect of α -mangostin on Ang II induced Vimentin mRNA levels in HK-2 cells; FIG. 4C is a graph of the effect of alpha-mangostin on Ang II-induced mRNA levels of alpha-SMA in HK-2 cells
FIG. 5 is a graph of the effect of alpha-mangostin on Ang II induced protein levels of the HK-2 EMT marker molecule; as compared with the normal group, the ratio of, # P＜0.05， ## p is less than 0.01; in comparison to the model set, * P＜0.05， ** p <0.01, n =3. Wherein FIG. 5A is a graph of the effect of alpha-mangostin on Ang II induced levels of Vimentin protein in HK-2 cells; FIG. 5B is a graph showing the effect of alpha-mangostin on Ang II induced levels of alpha-SMA protein in HK-2 cells; FIG. 5C is a graph showing the effect of alpha-mangostin on the levels of E-cadherin in Ang II-induced HK-2 cells
FIG. 6 is a graph showing the effect of α -mangostin on Ang II-induced expression of TGF- β receptors in HK-2 cells and their downstream Smad2 proteins; as compared with the normal group, the ratio of, # P＜0.05， ## p is less than 0.01; in comparison to the model set, * P＜0.05， ** p <0.01, n =3. Wherein FIG. 6A is a graph showing the effect of α -mangostin on Ang II-induced expression of TGF- β R1 protein in HK-2 cells; FIG. 6B is a graph of the effect of alpha-mangostin on Ang II-induced Smad2 protein expression in HK-2 cells
Detailed Description
The present invention is further explained with reference to the following specific examples, which are not intended to limit the present invention in any way.
Example 1: the compound alpha-mangostin improves the blood pressure of Spontaneous Hypertensive Rats (SHR)
Healthy male SHR rats (12-13 weeks old) and normal Wistar rats of the same week old purchased from beijing sbefu biotechnology limited, animal certification number: SCXK (Jing) 2019-0010. The test compound was prepared with 0.5% sodium carboxymethylcellulose (0.5% CMC-Na) and the volume administered was 10mL/kg for each group. SHR rats are randomly divided into a hypertension model group, a 0.5mg/kg and 1.0mg/kg dosage group of the inventive compound, a 20.0mg/kg dosage group of losartan, and normal Wistar rats of the same week age are normal control groups. Rats in the normal control group and the model control group are intragastrically administered with equal volume of solvent, and the positive drug and the compound group are intragastrically administered with losartan (20.0 mg/kg) and alpha-mangostin (0.5 and 1.0 mg/kg) respectively, once a day for 5 weeks. During the 1-3 week period, blood pressure determination training was performed on each rat for 30min at 9-12 am daily according to the standard operating instructions of a non-invasive rat tail blood pressure monitor (BP-2000 SERIES II) to adapt the animals to the blood pressure monitoring environment. Systolic, diastolic and mean blood pressures were monitored 1h (at 9-12 am) after dosing at weeks 3, 4 and 5, respectively.
Experiments show that: the systolic pressure, diastolic pressure and mean arterial pressure of spontaneous hypertension rats can be obviously reduced by intragastric administration of alpha-mangostin with a dose of 1.0mg/kg, the blood pressure reduction effect is equivalent to that of losartan with a dose of 20.0mg/kg (tables 1, 2 and 3), a strong blood pressure reduction effect is shown, the titer of the alpha-mangostin is better than that of a clinical blood pressure reduction drug losartan, and the alpha-mangostin is a novel blood pressure reduction drug and can be used for preparing oral blood pressure reduction drugs.
Table 1 effect of alpha-mangostin on mean blood pressure in Spontaneously Hypertensive Rats (SHR) (n =7-10,
)
note: in comparison with the normal group, ## P<0.01; in comparison to the set of models, * P<0.05， ** P<0.01. as a result, to
And (4) showing. Student's t-est was used for comparison between the two groups; comparisons among groups were performed using one-way analysis of variance (ANOVA), with LSD and Tamhane tests for all variances. The mean blood pressure is calculated as follows: mean blood pressure = systolic pressure/3 +2 × diastolic pressure/3
Table 2 effect of alpha-mangostin on systolic blood pressure in Spontaneously Hypertensive Rats (SHR) (n =7-10,
)
note: in comparison with the normal group, ## P<0.01; in comparison to the set of models, * P<0.05， ** P<0.01. the data statistics method is the same as that in Table 1.
Table 3 effect of alpha-mangostin on Spontaneous Hypertensive Rats (SHR) diastolic blood pressure (n =7-10,
)
note: as compared with the normal group, the test results, ## P<0.01; in comparison to the set of models, * P<0.05， ** P<0.01. the data statistics method is the same as that in Table 1.
Example 2: toxicity and safety test of compound alpha-mangostin
Healthy ICR mice of 20 animals with male and female halves and 18-22g weight are selected, the animals are fasted for about 12 hours before administration, the compound of the invention is prepared into suspension by 0.5 percent sodium carboxymethylcellulose, the suspension is intragastrically administered by 5g/kg according to the maximum concentration and the maximum volume of 40mL/kg body weight, the intragastrically administered is carried out once at 9 am, the administration is continuously observed for 14 days, and the poisoning and death conditions of the animals are recorded.
After the gavage administration, no obvious abnormal reaction occurs in animals, the animals are continuously observed for 14 days, no toxic reaction occurs in the animals, the animals can move freely, the animals can normally eat, drink, urinate and defecate, the weight of the animals normally increases, and the general condition is good. None of the 20 animals died, and after the animals were sacrificed, necropsy was observed, and no macroscopic pathological change was observed in each organ, and the maximum tolerance of the compound mouse by intragastric administration was determined to be more than 5g/kg.
Cell experiments show that the alpha-mangostin is less than 10 -5 mol/L had no effect on the viability of HK-2 cells.
Example 3: effect of the inventive Compound alpha-mangostin on the serum angiotensin II (AngII) and Nitric Oxide Synthase (NOS) in Spontaneously Hypertensive Rats (SHR)
The animals of the same batch are anesthetized by pentobarbital (50 mg/kg) after 5 weeks of administration and 1h of the last administration, blood is taken from abdominal aorta, centrifugation is carried out for 10min at 3600rpm, supernatant is taken, and the ELISA kit is used for detecting the levels of serum T-NOS, iNOS (Nanjing Indo) and AngII (Jiangsu enzyme-free).
As a result, ang ii was significantly increased in the model group compared to the normal group (P < 0.05) as shown in table 4. Compared with a model group, after the compound alpha-mangostin and the positive drug losartan are administrated for 5 weeks, the level of blood serum Ang II is obviously reduced, and the difference has statistical significance (P is less than 0.01); compared with the model group, the serum T-NOS level is obviously increased (P < 0.05) after the compound 1.0mg/kg and the positive medicament losartan are administrated for 5 weeks. The compound alpha-mangostin can inhibit the generation of angiotensin II (ANGII) and up-regulate total nitric oxide synthase (T-NOS), thereby reducing blood pressure and playing a role in preventing and treating hypertension.
Table 4 effect of alpha-mangostin on Ang ii and NOS in Spontaneous Hypertensive Rats (SHR) serum (n =6-8,
)
note: as compared with the normal group, the test results, # P<0.01; in comparison to the set of models, * P<0.05， ** P<0.01. the data statistics method is the same as that in Table 1.
Example 4: effect of alpha-mangostin on amelioration of renal injury induced by Spontaneously Hypertensive Rat (SHR)
Taking the same batch of animal serum, and detecting the levels of serum beta 2-MG (Jiangsu enzyme immune, MM-21246R 1), mAB (Jiangsu enzyme immune, MM-70785R 1) and NAG (Nanjing-built, A031-1-1) by using an ELISA kit. Rat kidney tissue is taken, 9 times volume of RIPA lysate is added, and the tissue is crushed by an ultrasonic crusher to prepare 10% homogenate. Centrifuging at 12000rpm/mi at 4 deg.C for 30min, collecting supernatant, quantifying with BCA protein quantification kit, and detecting protein expression levels of kidney EMT-related protein E-cadherin and alpha-SMA with western blotting.
The result is shown in figure 1, compared with the normal group, the urine beta 2-MG and NAG of the model group rats are obviously increased (P < 0.01), which indicates that the kidney of the SHR rats is damaged; compared with a model group, after the compound alpha-mangostin and the positive drug losartan are administrated for 5 weeks, urine beta 2-MG is remarkably reduced (P < 0.05), the dose group of the alpha-mangostin (0.5 MG/kg) remarkably reduces the urine NAG level (P < 0.05), and the urine mACLB level of rats of the alpha-mangostin (0.5 and 1.0 MG/kg) group has no obvious change (P > 0.05). The compound alpha-mangostin can improve the renal tubular injury caused by spontaneous hypertensive rats.
Western blotting results show (figure 2), compared with a normal group, the kidney E-cadherin protein expression level of the model group is obviously reduced (P < 0.05), and the alpha-SMA protein expression level is obviously increased (P < 0.05); compared with the model group, the compound alpha-mangostin (1.0 mg/kg) and the positive drug losartan group remarkably up-regulate the expression level of the E-cadherin protein (P < 0.05) and down-regulate the expression level of the alpha-SMA protein in the kidney (P < 0.05). The alpha-mangostin of the invention is suggested to improve renal fibrosis caused by spontaneous hypertensive rats.
Example 5: alpha mangostin has obvious improvement effect on Ang II-induced migration of HK2 cells and epithelial mesenchymal transformation, and the mechanism is probably related to inhibition of TGF-beta signal pathway.
1) Alpha-mangostin inhibits Ang II-induced HK-2 cell migration
Selecting HK-2 cells with good growth state, inoculating to a six-hole plate when the HK-2 cells grow to 70% -80% fusion, wherein the density of the seed plate is 1 × 10 5 Individual cells/per well, 37 ℃,5% CO 2 Incubating the cultured cells in a culture box, changing the medium containing 10% FBS to a serum-free low-sugar medium after the cells are attached to the walls, starving for 12h, sucking out the serum-free medium the next day, and adding alpha-mangostin (10) at each concentration -9 、10 -8 、10 -7 mol/L) and losartan (5X 10) -6 mol/L), 2mL per well, 1h apart, angII (10) -7 mol/L). Ang II induced HK-2 cells for 72 hours, during which the inducer or drug was replaced every 24 hours, and the cells were cultured in a cell culture chamber at 37 ℃. In the scratch experiment, the position of 0h is marked by a marker pen, a picture is taken, the picture is taken at the same position after 72h, the picture is compared before and after, and 3 points are taken for each cell. Mobility was calculated by calculating the migration area using Image Pro-Plus 6.0 software. As shown in the results (FIG. 3), the HK-2 cells induced by Ang II showed a significant increase in the mobility (P) in the model group as compared with the normal group<0.05 ); compared with the model group, the mobility of the alpha-mangostin and the positive drug losartan is obviously reduced (P)<0.05 It is suggested that the compound alpha-mangostin of the invention can inhibit the migration of HK-2 cells, thereby inhibiting the occurrence of EMT.
2) Effect of alpha-mangostin on mRNA of Ang II-induced HK-2 cell EMT marker molecule
After Ang II is induced for 24 hours, samples are collected, mRNA levels of renal tubular epithelial cell marker molecules E-cadherin, fibrosis marker molecules alpha-SMA and Vimentin are detected by q-PCR, and results are shown in (figure 4), compared with a normal group, the mRNA expression level of the model group E-cadherin is obviously reduced (P is less than 0.05), and the mRNA expression levels of the Vimentin and the alpha-SMA are obviously increased; compared with the model group, the alpha-mangostin and positive drug losartan group significantly increases the E-cadherin gene expression level (P < 0.05) and significantly reduces the Vimentin and alpha-SMA mRNA expression level (P < 0.05). The alpha-mangostin compound is suggested to be capable of obviously increasing E-cadherin and reducing Vimentin and alpha-SMA mRNA expression levels so as to inhibit the ANG II from inducing EMT of HK-2 cells.
3) Effect of alpha-mangostin on protein levels of Ang II-induced EMT marker molecules of HK-2 cells
48h of Ang II induction, changing the induction medicine and the culture medium once in the period of 24h, collecting samples after the end, detecting the protein levels of the epithelial marker molecule E-cadherin and the fibrosis marker molecule Vimentin and alpha-SMA of the HK-2 cells by Western blotting, and showing that the results are shown in figure 5, compared with the normal group, the expression levels of the Vimentin and the alpha-SMA protein of the model group are obviously increased (P is the expression level of the Vimentin and the alpha-SMA protein of the model group is obviously increased)<0.05 Marked down-regulation of E-cadherin protein expression levels (P)<0.05 ); compared with the model group, the alpha-mangostin and positive drug losartan group remarkably reduces the expression levels (P) of Vimentin and alpha-SMA protein<0.05 And alpha-mangostin (10) -9 mol/L) and the positive medicament losartan group obviously increase the expression level (P) of the E-cadherin protein<0.05). The alpha-mangostin is prompted to inhibit ANG II from inducing EMT of the HK-2 cells by inhibiting protein expression levels of fibrosis marker molecules Vimentin and alpha-SMA of the HK-2 cells induced by ANG II and increasing protein levels of epithelial marker molecules E-cadherin.
The results further show that the compound alpha-mangostin has a remarkable inhibition effect on Ang II induced EMT of HK-2 cells.
4) Effect of alpha-mangostin on Ang II-induced TGF-beta receptor of HK-2 cells and downstream Smad2 thereof
Selecting HK-2 cells with good growth state, inoculating to a six-hole plate when the HK-2 cells grow to 70% -80% fusion, wherein the density of the seed plate is 1 × 10 5 Individual cells/per well, 37 ℃,5% CO 2 Incubating the cultured cells in a culture box, changing the medium containing 10% FBS to a serum-free low-sugar medium after the cells are attached to the walls, starving for 12h, sucking out the serum-free medium the next day, and adding alpha-mangostin (10) at each concentration -9 、10 -8 、10 -7 mol/L) and losartan (5X 10) -6 mol/L), 2mL per well, 1h apart, angII (10) -7 mol/L). The HK-2 cells were harvested 24 hours after Ang II induction and Western blotting to measure protein levels of TGF-. Beta.R 1 and Smad2 signaling molecules downstream of the TGF-. Beta.signaling pathway, as shown in (FIG. 6): compared with the normal group, the expression level of TGF-beta R1 and Smad2 proteins in the model group is obviously increased (P)<0.05 ); compared with the model group, the alpha-mangostin and positive drug losartan group remarkably down-regulates the expression levels (P) of TGF-beta R1 and Smad2 proteins<0.05). The inventive compound alpha-mangostin is suggested to regulate the effect of Ang II induced fibrosis of HK-2 cells through a TGF-beta signal channel.
Furthermore, the medicine is a composition which takes alpha-mangostin as an active ingredient and contains auxiliary materials acceptable in the pharmaceutical field. Preferably, the medicament is an oral dosage form.
The recommended dosage of the alpha-mangostin for human bodies is 0.4-400 mg/kg.d.
Although the present invention has been described in detail with reference to the above embodiments, it should be understood by those skilled in the art that various changes may be made and equivalents may be substituted without departing from the spirit and scope of the invention, which is defined in the claims.
Claims (8)
1. Application of alpha-mangostin in preparing medicines for preventing and treating hypertension and/or hypertensive nephropathy.
2. The use of α -mangostin according to claim 1, wherein the hypertension is essential hypertension and the hypertensive renal disease is renal disease caused by essential hypertension.
3. The use of α -mangostin according to claim 1, wherein the α -mangostin inhibits angiotensin ii production and up-regulates total nitric oxide synthase, thereby lowering blood pressure and preventing and treating hypertension.
4. The use of α -mangostin according to claim 1, wherein α -mangostin improves renal tubular injury caused by hypertension by reducing the amount of NAG and β 2-MG in urine.
5. The use of α -mangostin according to claim 1, wherein α -mangostin up-regulates E-cadherin protein expression and down-regulates α -SMA protein expression, thereby improving renal fibrosis in spontaneously hypertensive rats.
6. The use of α -mangostin according to claim 1, wherein the medicament is a composition comprising α -mangostin as an active ingredient and pharmaceutically acceptable excipients.
7. The use of alpha-mangostin according to claim 6, wherein the medicament is an oral dosage form.
8. The use of alpha-mangostin according to claim 6, wherein the recommended dosage of alpha-mangostin for human use is 0.4-400 mg/kg.d.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| KR20150062583A (en) * | 2013-11-29 | 2015-06-08 | 동국대학교 산학협력단 | Composition containing alpha-mangosteen for preventing or treating non-alcoholic fatty liver disease and metabolic syndrome |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| KR20150062583A (en) * | 2013-11-29 | 2015-06-08 | 동국대학교 산학협력단 | Composition containing alpha-mangosteen for preventing or treating non-alcoholic fatty liver disease and metabolic syndrome |
Non-Patent Citations (1)
| Title |
|---|
| VIVIAN SOETIKNO等: "Alpha-Mangostin Improves Cardiac Hypertrophy and Fibrosis and Associated Biochemical Parameters in High-Fat/High-Glucose Diet and Low-Dose Streptozotocin Injection-Induced Type 2 Diabetic Rats", J EXP PHARMACOL. * |
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