Effect of internal heat acupuncture regulating TGF-β1/Smads signaling pathway on early healing of rotator cuff injury in rats
Objective To investigate the effect of acupuncture regulating TGF-β1/Smads signaling pathway on early healing of rotator cuff injury in rats.Methods A total of 72 8-week-old(200±20)g SPF SD male rats were divided into model group(54 rats)and control group(18 rats).In the model group,the rotator cuff injury rat model was constructed by disconnecting the right supraspinatus tendon of the rat,and control group was treated the same as model group except that supraspinatus tendon was not transversed.The rats with successful modeling were divided into rotator cuff injury group,internal heat acupuncture group,and internal heat acupuncture+pathway activator SRI-011381 group(internal heat acupuncture+SRI-011381 group)by simple random sampling,with 18 rats in each group.The internal heat acupuncture group was treated with internal heat acupuncture,internal heat acupuncture+SRI-011381 group was treated with intraperitoneal injection of 7 mg/kg SRI-011381 at the same time of internal heat acupuncture,control group and rotator cuff injury group were treated with ordinary acupuncture at the same time and in the same place as internal heat acupuncture group.The thermal withdrawal latency(TWL)of the right anterior full floor of rats was measured;the levels of interleukin(IL)-1β,IL-6,and tumor necrosis factor-α(TNF-α)were detected by enzyme-linked immunosorbent assay;HE staining was used to observe the pathological injury of rotator cuff tendon;Masson staining was used to observe the degree of rotator cuff joint tendon fibrosis;the maximum tensile load of rotator cuff was measured by tendine-bone interface biomechanics;the expression of tendon tissue related protein was detected by immunohistochemical staining;and the expression of TGF-β1/Smads signaling pathway was detected by Western blot.Results Compared with control group,TWL was shortened,maximum tensile load and the expression of Scx and TnC was decreased,the levels of IL-1 β,IL-6,and TNF-α,and the expression of TGF-β1,p-Smad2/3/Smad2/3 was increased in rotator cuff injury group(P＜0.05).Compared with rotator cuff injury group,TWL was longer,maximum tensile load and the expression of Scx and TnC was increased,the levels of IL-1 β,IL-6,and TNF-α,and the expression of TGF-β1,p-Smad2/3/Smad2/3 was reduced in internal heat acupuncture group(P＜0.05).Compared with internal heat acupuncture group,TWL was shortened,maximum tensile load and the expression of Scx and TnC was decreased,the levels of IL-1 β,IL-6,and TNF-α,and the expression of TGF-β1,p-Smad2/3/Smad2/3 was increased in internal heat acupuncture+SRI-011381 group(P＜0.05).In control group,all levels of tendon fiber,cartilage,and bone tissue were clearly arranged,collagen fiber generation was relatively normal,and blue collagen fiber deposition existed;in rotator cuff injury group,the tissue cell arrangement was disordered,inflammatory cell infiltration was obvious,collagen fiber production was relatively reduced,and blue collagen fiber deposition was reduced;in the internal heat acupuncture group,the histopathological changes were improved,cell arrangement was relatively normal,inflammatory cell infiltration was reduced,collagen fibers were abundant,blue collagen fiber deposition increased in a large area,and the continuity,parallel orientation,and density of collagen fibers increased;in internal heat acupuncture+SRI-011381 group,histopathological injury was aggravated,histopathic cell arrangement was disordered,inflammatory cell infiltration was aggravated,collagen fiber production was reduced,and blue collagen fiber deposition was significantly reduced.Conclusion Internal heat acupuncture can promote early healing of rotator cuff injury in rats,and its mechanism of action may be related to the inhibition of TGF-β1/Smads signaling pathway.
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