Electroacupuncture promotes gastrointestinal motility by activating autophagy of Cajal interstitial cells via downregulating PI3K/Akt/mTOR signaling pathway in stomach of diabetic gastro-paresis rats
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Electroacupuncture promotes gastrointestinal motility by activating autophagy of Cajal interstitial cells via downregulating PI3K/Akt/mTOR signaling pathway in stomach of diabetic gastro-paresis rats
ZHANG Tian-hua, ZHAO Sha-tong, LI Xiao-yu, et al. Electroacupuncture promotes gastrointestinal motility by activating autophagy of Cajal interstitial cells via downregulating PI3K/Akt/mTOR signaling pathway in stomach of diabetic gastro-paresis rats[J]. Acupuncture research, 2022, 47(12): 1060-1067.
DOI:
ZHANG Tian-hua, ZHAO Sha-tong, LI Xiao-yu, et al. Electroacupuncture promotes gastrointestinal motility by activating autophagy of Cajal interstitial cells via downregulating PI3K/Akt/mTOR signaling pathway in stomach of diabetic gastro-paresis rats[J]. Acupuncture research, 2022, 47(12): 1060-1067. DOI: 10.13702/j.1000-0607.20211241.
Electroacupuncture promotes gastrointestinal motility by activating autophagy of Cajal interstitial cells via downregulating PI3K/Akt/mTOR signaling pathway in stomach of diabetic gastro-paresis rats
0.01)。透射电镜结果显示,模型组、3-MA组大鼠胃窦部ICC细胞内受损较重,未见明显自噬结构;电针组、3-MA+电针组胃窦部ICC细胞内自噬小体数量明显增多,ICC受损状况有所改善。结论:电针可能通过调控PI3K/Akt/mTOR通路调节胃窦部ICC自噬水平,从而改善DGP胃肠动力功能障碍。Objective To observe the effect of electroacupuncture(EA) of “Zusanli”(ST36)
“Sanyinjiao”(SP6) and “Liangmen”(ST21) on gastrointestinal motility
blood glucose content and expression of autophagy-related proteins 1 light chain 3(LC3)
p62
phosphatidyli-nositol-3 kinase(PI3 K)
protein kinase B(Akt)
p-Akt and mammalian target protein of rapamycin(mTOR) of interstitial cells of Cajal(ICCs) in the cultured gastric antrum cells in diabetic gastroparesis(DGP) rats
so as to reveal its mechanisms underlying improvement of DGP. Methods A total of 45 Sprague Dawley(SD) rats were randomly divided into blank control
model
EA
medication(3-methyladenine
3-MA) and EA+3-MA groups
with 9 rats in each group. The DGP model was established by intraperitoneal injection of 2% streptozotocin(STZ) combined with high-fat and high sugar diet for 8 weeks. The gastric emptying rate was measured by using gavage of phenol red(to measure the propelling length of the phenol red/total length of small intestine ×100%). The symptom score(mental state
coat color and luster
behavior and activity
stool traits) of rats was observed every week and the blood glucose content was measured by using a glucometer. EA(20 Hz/100 Hz
2 mA) was applied to unilateral ST36
SP6 and ST21 alternatively for 15 min
once daily
5 days a week for 3 weeks. Rats of the 3-MA and 3-MA+EA groups received intraperitoneal injection of 3-MA(30 mg·kg(-1);3-MA+电针组予3-MA腹腔注射联合电针治疗;每个疗程5 d
Objective To observe the effect of electroacupuncture(EA) of “Zusanli”(ST36)
“Sanyinjiao”(SP6) and “Liangmen”(ST21) on gastrointestinal motility
blood glucose content and expression of autophagy-related proteins 1 light chain 3(LC3)
p62
phosphatidyli-nositol-3 kinase(PI3 K)
protein kinase B(Akt)
p-Akt and mammalian target protein of rapamycin(mTOR) of interstitial cells of Cajal(ICCs) in the cultured gastric antrum cells in diabetic gastroparesis(DGP) rats
so as to reveal its mechanisms underlying improvement of DGP. Methods A total of 45 Sprague Dawley(SD) rats were randomly divided into blank control
model
EA
medication(3-methyladenine
3-MA) and EA+3-MA groups
with 9 rats in each group. The DGP model was established by intraperitoneal injection of 2% streptozotocin(STZ) combined with high-fat and high sugar diet for 8 weeks. The gastric emptying rate was measured by using gavage of phenol red(to measure the propelling length of the phenol red/total length of small intestine ×100%). The symptom score(mental state
coat color and luster
behavior and activity
stool traits) of rats was observed every week and the blood glucose content was measured by using a glucometer. EA(20 Hz/100 Hz
2 mA) was applied to unilateral ST36
SP6 and ST21 alternatively for 15 min
once daily
5 days a week for 3 weeks. Rats of the 3-MA and 3-MA+EA groups received intraperitoneal injection of 3-MA(30 mg·kg
(-1)
·d(-1)·d
(-1)
10 mg/mL)
once daily
5 days a week for 3 weeks. After 15 days' intervention
the rats were operated for gastric emptying rate test
specimen collection
isolation
and culture of primary ICCs. The expression levels of microtubule associated protein LC3
p62
PI3 K
Akt
p-Akt and mTOR of ICCs of cultured gastric antrum cells were detected using Western blot
and the number of autophagosomes in ICC of gastric antrum was observed under transmission electron microscope. Results Compared with the blank control group
the symptom score
blood glucose
and the expression levels of p62
class Ⅰ PI3 K
Akt
p-Akt and mTOR proteins were increased significantly(P
<
0.01)
while the gastric emptying rate and ratio of LC3Ⅱ/LC3Ⅰ and the expression level of class Ⅲ PI3 K protein were significantly decreased(P
<
0.05
P
<
0.01) in the model group. In comparison with the model group
the increase of symptom score
blood glucose
and expression levels of p62
class Ⅰ PI3 K
Akt
p-Akt and mTOR proteins and the decrease of gastric empty rate and LC3Ⅱ/LC3Ⅰ ratio and the expression level of class Ⅲ PI3 K protein were all reversed in both EA and EA+3-MA groups(P
<
0.05
P
<
0.01)
rather than in the 3-MA group. In addition
3-MA also reversed modeling-induced increase of class Ⅰ PI3 K
Akt
p-Akt and mTOR proteins expression(P
<
0.01). No significant differences were found between the EA and EA+3-MA in downregulating the levels of symptom score and blood glucose content
and in upregulating gastric empty rate(P
>
0.05). The effect of EA was notably superior to that of EA+3-MA in upregulating the ratio of LC3Ⅱ/LC3Ⅰ and the expression level of class Ⅲ PI3 K protein
and in downregulating the expression of p62
class Ⅰ PI3 K
Akt
p-Akt and mTOR proteins(P
<
0.05
P
<
0.01). The findings of transmission electron microscopy showed obvious swelling
breakage of some mitochondrial cristae in the ICC cells of antrum and no autophagosomes in the model group and 3-MA group
which was milder in the damage of mitochondrial cristae and marked increase in the autophagosomes in both EA and EA+3-MA groups. Conclusion EA can improve the gastrointestinal motility and symptoms in DGP rats
which may be related to its functions in downregulating PI3 K/Akt/mTOR signaling to promote autophagy level of ICC.
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